Antioxidant, anti-prostate cancer potential, and phytochemical composition of the ethyl acetate stem bark extract of Boascia coriacea (Pax.)

Gervason Moriasi; Mathew Ngugi; Peter Mwitari; George Omwenga

doi:10.1371/journal.pone.0309258

Abstract

Background

The antioxidant and anticancer potential of natural compounds, particularly from medicinal plants, is increasingly being explored as alternatives to synthetic antioxidants and chemotherapeutics. Boascia coriacea (Pax) has been traditionally used for treating various ailments, including oxidative stress-related diseases and prostate cancer. However, there is a paucity of empirical evidence to validate the ethnomedicinal claims, hence this study.

Methods

The antioxidant capacity of the extract was assessed using 1,1-Diphenyl-2-picryl Hydrazyl radical (DPPH) radical scavenging and hydrogen peroxide scavenging assays, alongside total antioxidant capacity. In vitro cytotoxicity was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on Vero CCL-81 normal cells and DU-145 prostate cancer cells. Gene expression levels of ar, bcl-2, caspase 3, cdk1, and p53 were quantified using qPCR to elucidate the mechanisms of action. Phytochemical analysis was conducted using gas chromatography-mass spectrometry.

Results

The studied plant extract exhibited significant DPPH radical scavenging activity, with an EC₅₀ of 0.008 μg/ml, 10-fold lower than that of L-ascorbic acid (0.08 μg/ml), indicating potent antioxidant capacity. Similarly, the extract demonstrated substantial hydrogen peroxide scavenging activity, albeit with lower efficacy (EC₅₀ of 1039.10 μg/ml) compared to L-ascorbic acid, and a total antioxidant capacity of 454.39±25.26 μg AAE/mg dw. In vitro cytotoxicity assay revealed a CC₅₀ of 68.61 μg/ml against Vero CCL-81 cells and an IC₅₀ of 32.16 μg/ml against DU-145 cells, with a superior selectivity index of 2.13, compared to doxorubicin’s 1.46. The extract significantly downregulated the expression of ar, bcl-2, normalised caspase 3, cdk1 genes while upregulating p53 in DU-145 cells, suggesting its role in inducing apoptosis and inhibiting cancer cell proliferation. Phytochemical analysis identified 19 compounds, including lup-20(29)-en-3-one (7.99%) and lupeol (59.49%), which are associated with anticancer activity.

Conclusion

The ethyl acetate stem bark extract of B. coriacea demonstrates significant antioxidant and anticancer activities, potentially through modulation of apoptosis and cell cycle pathways. The presence of bioactive compounds supports its potential as a therapeutic agent, warranting further investigation for developing novel treatments for prostate cancer and oxidative stress-related conditions.

Citation: Moriasi G, Ngugi M, Mwitari P, Omwenga G (2024) Antioxidant, anti-prostate cancer potential, and phytochemical composition of the ethyl acetate stem bark extract of Boascia coriacea (Pax.). PLoS ONE 19(12): e0309258. https://doi.org/10.1371/journal.pone.0309258

Editor: Akingbolabo Daniel Ogunlakin, Bowen University, NIGERIA

Received: August 7, 2024; Accepted: October 26, 2024; Published: December 2, 2024

Copyright: © 2024 Moriasi et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Data Availability: All relevant data are within the manuscript and its Supporting information files.

Funding: This study was partly supported by the German Academic Exchange (DAAD) scholarship for PhD Study granted to Gervason Moriasi under the In-Country/In-Region Scholarship Programme (Ref: 91843013). There was no additional external funding received for this study.

Competing interests: The authors have declared that no competing interests exist.

1. Introduction

Oxidative stress (OS), is a fundamental factor in the pathogenesis of numerous debilitating conditions, including PCa [1]. The body’s intrinsic enzymatic and nonenzymatic antioxidant defence mechanisms typically counterbalance these pro-oxidants to maintain redox homeostasis. However, during disease states or excessive exposure to pro-oxidants, these defences are overwhelmed, leading to sustained oxidative stress and the progression of various diseases, including cancer [2]. Synthetic antioxidants such as butylated hydroxyanisole (BHA), tertiary butylhydroquinone (TBHQ), butylated hydroxytoluene (BHT), and propyl gallate (PG) are currently employed to bolster endogenous defences against OS [3]. Despite their effectiveness in scavenging free radicals, their extensive use has been linked to adverse health effects, including carcinogenicity, cytotoxicity, endocrine disruptions, and immunological disorders. Given the significant role of oxidative stress in prostate cancer (PCa) and the limitations of synthetic antioxidants, there is a critical need for alternative antioxidants from natural sources that are both safe and effective in mitigating OS and its associated pathologies.

Cancer remains one of the leading causes of mortality worldwide, ranking as either the first or second leading cause of death in most countries. Patients often succumb to the disease prematurely, before the age of 70 years [4]. Prostate cancer, in particular, present significant health challenges, with high morbidity and mortality rates affecting across the globe [5]. Sub-Saharan African countries, including Kenya, bear a disproportionate cancer burden, with over 80% of cancer cases attributed to limited access to quality healthcare In Kenya, cancer is the second leading cause of disease-related deaths, following cardiovascular disorders [6]. Besides, research indicates a higher mortality rate among PCa patients of African descent compared to their white counterparts, influenced by genetic, socioeconomic, and environmental factors [7]. This disparity is largely due to ineffective healthcare systems, the high cost of cancer treatment, and limited resources for accessing quality care [8]. Additionally, conventional anticancer therapies often lead to severe side effects, therapeutic failure, and high recurrence rates [9], underscoring the urgent need for alternative, safer, and more affordable treatments.

Boscia coriacea (Pax), a woody shrub native to Kenya and known locally by various names, is an important plant in traditional medicine and ethnoveterinary practices. Its stem bark, roots, and leaves are used to treat ailments such as body pains, ear problems, and respiratory infections, as well as veterinary conditions like anthrax [10]. Recent phytochemical studies have identified several bioactive compounds in B. coriacea, including alkaloids, flavonoids, glycosides, and tannins, which exhibit antimicrobial, anti-inflammatory, antioxidant, and anticancer activities [10]. However, comprehensive scientific studies on its antioxidant and anticancer properties, safety, and detailed phytochemical profile are limited. Therefore, this study aimed to investigate the cytotoxicity, anti-prostate cancer, antioxidant activity, and phytochemical composition of the ethyl acetate stem bark extract of B. coriacea, thereby validating its traditional uses and exploring its potential as a source of new therapeutic agents against oxidative stress and associated diseases, such as prostate cancer.

2. Materials and methods

2.1 Plant material

Fresh stem barks of mature B. coriacea were collected from their natural habitat in Mbeere North Sub-County, Embu County, Kenya as described by previously [11]. The selection of this plant for this study was based on its ethnomedicinal use by the local community for managing disorders associated with oxidative stress, especially cancer of the prostate. Initially, this plant was identified by a local herbalist by its local name as "Mutangira”. The plants were taxonomically authenticated by a qualified botanist (NMK/BOT/CTX1/2), and duplicate specimens archived at the Department of Botany, National Museums of Kenya, for future reference. The collected plant material was chopped into small pieces and spread on a wooden bench to dry naturally in a well-ventilated room for two weeks. Once dried, the material was ground into a powder using an electric plant grinder, and the resulting powder was packed and stored in a labelled khaki envelope pending extraction.

2.2 Extraction

The cold maceration method described previously [12] was employed following sight modifications [13]. Briefly, 100 grams of the powdered plant material were macerated in 1000 millilitres of analytical grade ethyl acetate in a well-labelled conical flask for 48 hours, with occasional agitation using a mechanical shaker. The resulting mixture was decanted and filtered through Whatman No. 1 filter paper into a separate flask. And concentrated in vacuo at 50°C using a rotary evaporator. The extract was further dried at 30°C for five days in a hot air oven, and percentage yield was computed according to Eq 1 [14].

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2.3 Determination of in vitro antioxidant effects of the study plant extract

2.3.1 Determination of 1,1-Diphenyl-2-picryl Hydrazyl Radical (DPPH) scavenging activity.

The experimental procedure was adapted from an established method [15] with slight modifications [13]. A 0.3 mM DPPH solution was prepared by dissolving 12 mg of 1,1-diphenyl-2-picrylhydrazyl in 100 ml of analytical-grade methanol. Test samples, including the extract and ascorbic acid, were prepared at concentrations of 1000, 100, 10, 1, 0.1, and 0.01 μg/ml. In triplicate, 1 ml of each test sample was mixed with 2.5 ml of the DPPH solution and incubated in the dark for 15 minutes. Absorbance was measured at 517 nm using a Shimadzu UV-VIS 1600 spectrophotometer within 60 minutes, against a control. The percentage of DPPH radical scavenging activity (% RSA) was calculated using Eq 2, and the EC₅₀ values, indicating the concentration required to scavenge 50% of DPPH radicals, were determined using linear regression plot of % RSA versus concentration to evaluate antioxidant activity.

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2.3.2 Hydrogen peroxide scavenging activity.

The hydrogen peroxide scavenging activity (HPSA) of the plant extract was evaluated using a modified protocol [16]. Briefly, 600 μl of 40 mM hydrogen peroxide in 0.1 M phosphate buffer (pH 7.4) was mixed with 1 ml of the plant extract or L-ascorbic acid across a concentration range of 0.01–1000 μg/ml. Following a 10-minute incubation at room temperature, absorbance was measured at 230 nm with a Shimadzu UV-Vis 1600 double-beam spectrophotometer, using phosphate buffer as the blank. The percentage HPSA was calculated according to Eq 3 [16], and the EC₅₀ value, representing the concentration required to scavenge 50% of hydrogen peroxide, was determined from a linear regression plot of % HPSA versus concentration.

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2.3.3 Determination of total antioxidant capacity.

In this experiment, we employed a modified phosphomolybdenum method [17] to assess the extract’s total antioxidant capacity. L-ascorbic acid concentrations (0.01 to 1000 μg/ml) was prepared and added in triplicate to a reaction mixture containing 0.6 M sulfuric acid, 28 mM sodium phosphate, and 4 mM ammonium molybdate. The samples were incubated at 95°C for 90 minutes, and cooled to room temperature, and their absorbances were recorded at 695 nm using a Shimadzu UV-Vis 1600 double-beam spectrophotometer, with 300 μl methanol serving as the blank. A linear regression curve of absorbance versus L-ascorbic acid concentration was generated to interpolate the ascorbic acid concentrations in plant extracts. Each plant extract (300 μl) was treated identically, with absorbance measured at 695 nm. All assays were performed in triplicate. The total antioxidant capacity of the extract was quantified as ascorbic acid equivalent per milligram of dry weight (μg AAE/mg d.w), providing a measure of antioxidant capacity.

2.4 Determination of in vitro cytotoxic effects of the study extract against normal mammalian and PCa cells

2.4.1 Preparation of plant extracts.

The plant extract was precisely weighed (100 mg), dissolved in dimethyl sulphoxide (DMSO) (10 ml), and thoroughly vortexed. Subsequently, 100 μl of the solubilized extract was diluted with phosphate-buffered saline (PBS) to a final volume of 1000 μl, achieving a stock concentration of 1000 μg/ml. Serial dilutions using a three-fold factor produced seven concentrations: 1.37 μg/ml, 4.12 μg/ml, 12.35 μg/ml, 37.04 μg/ml, 111.11 μg/ml, 333.33 μg/ml, and 1000 μg/ml of the extract. The reference drug, Doxorubicin, was similarly prepared, yielding concentrations of 0.04 μg/ml, 0.12 μg/ml, 0.37 μg/ml, 1.11 μg/ml, 3.33 μg/ml, 10 μg/ml, and 30 μg/ml.

2.4.2 Cell culture and maintenance.

The cell lines were cultured and maintained using a standard protocol [18]. Vero CCL-81 cells were grown in Eagle’s Minimum Essential Medium (EMEM) supplemented with 10% foetal bovine serum (FBS), 2 mM L-glutamine, and antibiotics (100 U/ml penicillin and 0.1 mg/ml streptomycin). DU-145 cells were cultured in high-glucose Dulbecco’s Modified Eagle Medium (DMEM) with similar supplements. All cultures were incubated at 37°C in a humidified incubator supplied with 5% CO₂ for 48 hours. Cells were routinely monitored, trypsinised, and passaged, and those at ≥90% confluence were harvested and prepared for subsequent assays.

2.4.3 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cytotoxicity and cancer cell inhibition.

The cytotoxicity of plant extract on normal mammalian cells (Vero CCL-81) was evaluated using the MTT assay [19] with slight modifications [18]. Cells were seeded at a density of 2×10⁴ cells/well in 96-well plates with 100 μL of medium. Following overnight incubation at 37°C with 5% CO₂, the cells were treated in triplicate with serial dilutions of plant extracts (0–1000 μg/mL in 0.1% DMSO) and the reference drug, Doxorubicin. After 48 hours of incubation under the same conditions, 10 μL of freshly prepared MTT reagent (5 mg/mL in PBS) was added to each well, and plates were incubated for an additional 4 hours, as detailed in section 2.4.2. Supernatants were aspirated, replaced with 100 μL of DMSO (0.1%), and agitated. Then, absorbance was measured at 570 nm and used to calculate cytotoxicity/inhibition percentages according to Eq 4.

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Furthermore, the median cytotoxic concentrations (CC₅₀) for Vero CCL-81 cells and median inhibitory concentrations (IC₅₀) for DU-145 cells were derived from a linear regression plot of percentage cytotoxicity/inhibition versus concentration to evaluate the safety of the extract. Moreover, the selectivity index (SI) of the plant extract was calculated using Eq 5, described previously [20].

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2.4.4 Determination of the expression profiles of cancer-associated genes in the DU-145 cells treated with the selected plant extract.

The expression profiles of key genes involved in cancer initiation and progression were analysed using quantitative real-time PCR (qPCR). DU-145 cells (1×10⁶) were seeded in 96-well plates and treated with the plant extract at a concentration corresponding to its IC₅₀ and upon the determination of its selectivity, based on SI value, followed by a 48-hour incubation at conditions described in section 2.4.2. Total RNA was extracted using a miniprep kit (Solis BioDyne) and quantified with a nanodrop spectrophotometer (ThermoFisher Scientific). RNA (2 μg) was reverse transcribed into cDNA using a cDNA synthesis kit (Solis BioDyne), added SYBR Green dye, and then amplified using specific primers (Table 1) for Bcl2, ar, cdk1, p53, caspase 3, with actb and gapdh as the normalisation controls. Gene expression was measured with a QuantStudio^™ 5 System (ThermoFisher Scientific) and analysed using the comparative threshold (CT) method, with fold changes calculated via the 2^-ΔΔCt approach [21].

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Table 1. Target genes and their respective primers.

https://doi.org/10.1371/journal.pone.0309258.t001

2.5 Phytochemical analysis

Phytochemical analysis was conducted using gas chromatography-mass spectrometry (GC-MS), following a standard protocol [22]. Analytical grade ethyl acetate (Sigma-Aldrich) was used. Standard reference compounds for GC-MS calibration and comparison were also obtained from Sigma-Aldrich. The ethyl acetate stem bark extract of B. coriacea was dissolved using its own solvent to get a concentration of 1 mg/ml. The sample solution was filtered through a 0.45μm PTFE syringe filter, then injected into a Shimadzu QP 2010-SE GC-MS equipped with an auto-sampler. Ultrapure helium (He) served as the carrier gas at a linear velocity of 35 cm/s. Separation was achieved using a BPX5 nonpolar column (30 m × 0.25 mm ID; 0.25 μm film thickness). The GC oven was programmed from 60°C, ramping at 10°C/min to 200°C (hold for 1 min), then at 10°C/min to 280°C (hold for 10 min), totalling a runtime of 33 minutes. The injection temperature was 250°C, the interface temperature was 250°C, and the EI ion source operated at 200°C in electron impact (EI) mode at 70 eV. Mass spectrometry was conducted in Scan mode over an m/z range of 35–550 a.m.u., with sample injection in split mode at a split ratio of 10:1.

2.6 Data management, statistical analysis, and reporting

In vitro antioxidant, cytotoxicity, and antiproliferative data were initially organized in Microsoft Excel (Microsoft 365) and subsequently analysed using Minitab software version 21.4. Descriptive statistics were computed, and results were expressed as mean ± standard deviation () from replicate experiments. Normality of the quantitative data from antioxidant and cytotoxicity experiments was assessed using the Shapiro-Wilk test, and deemed normally distributed. Afterward, statistical comparisons among means were conducted using unpaired student t-test or One-Way Analysis of Variance (ANOVA) with Tukey’s post hoc test at a significance level of α = 0.001, and graphical presentation of the findings was performed using GraphPad Prism version 10 software. Gene expression data were analysed using the 2^-ΔΔCt method via the QuantStudio^™ 5 System’s software. Compounds were identified by matching mass spectra against the NIST library and calculating retention indices with a homologous series of n-alkanes (C8-C20) under the same GC-MS conditions. Verification involved comparing mass spectra and retention indices with standards on the National Library of Medicine’s PubChem (nih.gov), NIST Chemistry WebBook, and scholarly literature. The identified compounds were catalogued by name, class, molecular weight, molecular formula, and retention time, with their abundance determined from peak areas in the total ion chromatogram. Agilent ChemStation software was used for data analysis.

2.7 Ethics approval and consent to participate

The present study and all its protocols were approved by the Kenyatta University Ethics Review Committee (reference number: PKU/2652/11787) and the National Commission for Science, Technology, and Innovation (reference number: NACOSTI/P/23/25162). Consent to participate was not required for this study.

3. Results

3.1 In vitro antioxidant activity

3.1.1 DPPH radical scavenging activity of stem bark extracts of B. coriacea.

The evaluation of DPPH radical scavenging activity of the ethyl acetate stem bark extract of B. coriacea revealed no significant differences at concentrations of 0.01 μg/ml, 0.1 μg/ml, and 1 μg/ml, as well as at 10 μg/ml and 100 μg/ml (P>0.001; Fig 1; S1 File). However, a significant increase in percentage scavenging activity was observed with increasing concentrations, (P<0.001; Fig 1; S1 File). Besides at concentrations of 0.01 μg/ml and 0.1 μg/ml, the ethyl acetate stem bark extract of B. coriacea showed significantly higher DPPH radical scavenging activities than those of L-ascorbic acid (P<0.001; Fig 1; S1 File). At a concentration of 1 μg/ml, no significant difference between the DPPH radical scavenging activity of the studied plant extract and L-ascorbic acid (P>0.001; Fig 1; S1 File). However, at concentrations of 10 μg/ml, 100 μg/ml, and 1000 μg/ml, L-ascorbic acid exhibited significantly higher DPPH radical scavenging activities than the ethyl acetate stem bark extract of B. coriacea (P<0.001; Fig 1; S1 File).

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Fig 1. In vitro DPPH radical scavenging effects of ethyl acetate stem bark extract of B. coriacea.

The results are presented as for triplicate experiments. Bars with different subscript lowercase letters within the same concentration are significantly different (P<0.0001; Unpaired student t-test) and those with different uppercase superscript letters across concentrations are significantly different (P<0.001; One-Way ANOVA with Tukey’s post hoc test); % DPPH RSA: Percentage 1,1-Diphenyl-2-picryl Hydrazyl Radical Scavenging Activity; BC-EA: Ethyl acetate stem bark extract of B. coriacea; L-ASA: L-Ascorbic Acid.

https://doi.org/10.1371/journal.pone.0309258.g001

Furthermore, it was observed that the EC₅₀ value of the study plant extract (0.008 μg/ml) was tenfold lower compared to hat L-ascorbic acid (0.08 μg/ml) (Fig 1).

3.1.2 Hydrogen peroxide scavenging activity of the stem bark extracts of B. coriacea.

The ethyl acetate stem bark extract of B. coriacea displayed no significant differences in hydrogen peroxide scavenging activity at concentrations of 1 μg/ml to 1000 μg/ml (P > 0.001), though these activities were markedly higher than those observed at 0.01 μg/ml and 0.1 μg/ml (P < 0.001; Fig 2; S2 File). Likewise, the hydrogen peroxide scavenging activities of the ethyl acetate stem bark extract of B. coriacea at concentrations of 0.1 μg/ml to 10 μg/ml were not significantly different (P>0.001; Fig 2; S2 File). Notably, the hydrogen peroxide scavenging activity of L-ascorbic acid increased significantly with increasing concentration (P<0.001; Fig 2; S2 File).

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Fig 2. Hydrogen peroxide scavenging activity of the ethyl acetate stem bark extract of B. coriacea.

The results are presented as for triplicate experiments. Bars with different subscript lowercase letters within the same concentration are significantly different (P<0.0001; Unpaired student t-test) and those with different uppercase superscript letters across concentrations are significantly different (P<0.001; One-Way ANOVA with Tukey’s post hoc test); BC-EA: Ethyl acetate stem bark extract of B. coriacea; L-ASA: L-Ascorbic Acid.

https://doi.org/10.1371/journal.pone.0309258.g002

At all the tested concentrations, the ethyl acetate stem bark extract of B. coriacea showed significantly lower hydrogen peroxide scavenging activity than L-ascorbic acid (P<0.001; Fig 2; S2 File). Additionally, the ethyl acetate extract had a considerably higher EC₅₀ (1039.10 μg/ml) than L-ascorbic acid (Fig 2).

3.1.3 Total antioxidant capacity of the stem bark extracts of B. coriacea.

This study showed that the total antioxidant capacity of the ethyl acetate stem bark extract of B. coriacea was high at 454.39±25.26 μg AAE /mg dw.

3.2 In vitro cytotoxicity and antiproliferative effects

3.2.1 In vitro cytotoxicity of the ethyl acetate stem bark extract of B. coriacea.

Overall, the ethyl acetate stem bark extract of B. coriacea exhibited positive significant (P<0.001) concentration-dependent cytotoxicity against Vero CCL-81 cells (Fig 3; S3 File). No significant difference between the cytotoxicity of the studied plant extract at concentrations of 333.33 μg/ml and 1000 μg/ml, and among those observed at concentrations of 12.35 μg/ml, 37.04 μg/ml, and 111.11 μg/ml, respectively (P>0.001; Fig 3; S3 File). Besides, the cytotoxicity of doxorubicin at concentrations of 3.33 μg/ml, 10.00 μg/ml, and 30.00 μg/ml (P>0.001), between 1.11 μg/ml and 3.33 μg/ml (P>0.001), 0.12 μg/ml and 0.37 μg/ml (P>0.001), and 0.04 μg/ml and 0.12 μg/ml (P>0.001), respectively, was not significantly different (Fig 3; S3 File). Furthermore, the CC₅₀ of the ethyl acetate extract was 68.61 μg/ml, and higher than that of doxorubicin (0.41 μg/ml) as shown in Fig 3.

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Fig 3. Cytotoxic effects of the ethyl acetate stem bark extract of B. coriacea against Vero CCL-81 cells.

The results are presented as for four triplicate experiments. Different uppercase letters across concentrations are significantly different (P<0.001; One-Way ANOVA with Tukey’s post hoc test); BC-EA: Ethyl acetate stem bark extract of B. coriacea; CC₅₀: Median cytotoxic concentration.

https://doi.org/10.1371/journal.pone.0309258.g003

3.2.2 Effects of the ethyl acetate stem bark extract of B. coriacea on DU-145 cells.

The inhibitory effects of the ethyl acetate stem bark extracts of B. coriacea against DU-145 cells did not differ significantly at concentrations of between 1.37 and 12.35 μg/ml and between 111.11 and 333.33 μg/ml (P>0.001; Fig 4; S4 File). Also, the difference between the inhibitory effects of the studied plant extract at concentrations of between 37.04 μg/ml and 111.11 μg/ml were insignificant (P>0.001; Fig 4; S4 File). Nevertheless, the percentage inhibition of DU-145 by this extract increased significantly with increasing extract concentration (P<0.001; Fig 4; S4 File). Besides, no significant difference was observed between the inhibition of DU-145 cells by doxorubicin at concentrations of 10 μg/ml and 30 μg/ml, was observed (P>0.001; Fig 4; S4 File); however, the inhibitions recorded at lower concentrations increased significantly with increasing concentration (P<0.001; Fig 4; S4 File). Moreover, the IC₅₀ of this extract was 32.16 μg/ml and higher than that of the reference drug, doxorubicin (0.28 μg/ml) (Fig 4).

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Fig 4. Anti-PCa effects of the ethyl acetate stem bark extract of B. coriacea against DU-145 cells.

The results are presented as for four triplicate experiments. Different uppercase letters across concentrations are significantly different (P<0.001; One-Way ANOVA with Tukey’s post hoc test); BC-EA: Ethyl acetate stem bark extract of B. coriacea; CC₅₀: Median cytotoxic concentration.

https://doi.org/10.1371/journal.pone.0309258.g004

3.2.3 Selectivity index.

The selectivity index (SI) of the studied plant extract was computed to determine its ability to selectively exert cytotoxic effects on cancer cells while sparing normal cells. The results showed that the ethyl acetate stem bark extract of B. coriacea was 2.13 and relatively higher than that of doxorubicin (1.46).

3.3 Expression levels of selected genes in DU-145 cells treated with the selected plant extracts

It was also observed that the DU-145 cells treated with the ethyl acetate stem bark extract of B. coriacea showed a significantly lower expression of the ar, bcl₂, caspase 3 and cdk1 genes (P<0.001; Table 2). Notably, the expression of p53 gene was significantly higher in DU-145 treated with the studied plant extract than in the control cells (P<0.001; Table 2).

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Table 2. Target gene expression profile in DU-145 cells treated with the ethyl acetate stem bark extract of B. coriacea.

https://doi.org/10.1371/journal.pone.0309258.t002

3.4 Phytochemical compounds detected in the ethyl acetate stem bark extract of B. coriacea

Phytochemical analysis of the ethyl acetate (BC-EA) showed the presence of 19 compounds distributed in various groups of chemical classes including glyceride esters, glycerides, hydantoins, phenolic aldehydes, alkenes, phthalates, aliphatic alcohol fatty acids, esters, diterpenoids, as well as aliphatic amine, alkylated alkanes, and phytosterols. Notably, Lup-20(29)-en-3-one (7.99%) and Lupeol (59.49%) were the most dominant (Table 3). Conversely, 4-Hydroxy-2-methoxybenaldehyde (0.22%) and 1-Octadecene (2.50%) were the least abundant compounds in this extract (Table 3). The obtained chromatograph is presented in Fig 5.

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Fig 5. A GC-MS chromatogram showing peaks of the detected phytochemical compounds in the ethyl acetate stem bark extract of B. coriacea.

https://doi.org/10.1371/journal.pone.0309258.g005

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Table 3. Phytochemicals identified in the ethyl acetate stem bark extract of B. coriacea.

https://doi.org/10.1371/journal.pone.0309258.t003

4. Discussion

The antioxidant research landscape has undergone a significant transformation in recent years, with a growing emphasis on identifying natural compounds that can mitigate oxidative stress and its associated disorders, including cancer, diabetes, and neurodegenerative disorders [23]. Synthetic antioxidants, such as BHA and BHT, cause undesirable effects, including cancer and autoimmunity, limiting their clinical significance. Contrastingly, natural antioxidants derived from medicinal plants are considered safer and more effective [24]. Therefore, the antioxidant potential of the ethyl acetate stem bark extract of B. coriacea was investigated to appraise its potential in averting OS and associated complications, especially cancer of the prostate.

The DPPH assay technique is reputed for its accuracy and reproducibility in measuring the ability of a compound to scavenge DPPH radicals, leading to a colour change from purple/violet to yellow, depending on the concentration of the antioxidant, and a reduction in absorbance measured at 517 nm [25]. The results showed that the ethyl acetate stem bark extract of B. coriacea exhibited significant DPPH radical scavenging activity, with a concentration-dependent increase in efficacy. This suggests that the extract contains robust antioxidants, emphasizing its potential as a putative source of antioxidant compounds [26].

The hydrogen peroxide scavenging activity of the ethyl acetate stem bark extract of B. coriacea was also investigated using a previously described procedure [27]. Hydrogen peroxide is a significant ROS that can penetrate cells and induce DNA strand breaks, leading to genetic instability and various disorders, including cancer [28]. The results showed that the studied lant extract possesses considerable hydrogen peroxide scavenging efficacy, suggesting its potential in preventing the generation of hydroxyl radicals and their devastating effects on the cell.

The total antioxidant capacity (TAC) of the ethyl acetate stem bark extract of B. coriacea was evaluated using a previously established method [17]. The results showed that the extract possessed significant TAC, indicating its potential in preventing and alleviating oxidative stress [29]. The observed TAC further typifies the various antioxidant effects reported in this study, underscoring the potential of the studied plant extract as a promising source of potent antioxidant compounds which can be optimized to avert oxidative stress.

The quest for effective and safe anticancer therapies remains a pressing concern, particularly in less-developed countries where cancer burden is disproportionately high (~ 80%) exacerbated by limited healthcare resources [4]. In this context, the exploration of plant-derived compounds as potential sources of novel anticancer agents is a feasible and viable alternative stratagem to identify, optimise, and develop efficacious, safe, and cost-effective armamentaria. The cytotoxic and anti-PCa effects of the ethyl acetate stem bark extract of B. coriacea (Pax.) were investigated in this study to appraise its efficacy.

The in vitro cytotoxic effects of the studied plant extract on normal mammalian cells (Vero CCL-81) and cancer cell line (DU-145) were investigated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay technique [18]. This method is renowned for its high sensitivity, reliability, and reproducibility, making it an ideal tool for assessing cell viability in drug toxicity and anticancer efficacy studies [30]. In the present investigation, the ethyl acetate stem bark extract of B. coriacea exhibited concentration-dependent cytotoxicity with moderate efficacy (30 μg/ml < CC₅₀ < 100 μg/ml) towards Vero CCL-81 cells, according to the criteria stipulated by the National Cancer Institute [31]. This concentration-dependent increase in cytotoxicity can be attributed to the increasing concentration of certain toxic phytochemicals [32].

The results further demonstrated significant extract-dependent increases in percentage inhibitions of growth of cancer cells, with the ethyl acetate stem bark extract of B. coriacea showing remarkable antiproliferative efficacy (CC₅₀ < 30 μg/ml). This finding is consistent with previous reports indicating concentration-dependent antiproliferative effects of plant extracts on cell lines [33]. The observed antiproliferative effects of the ethyl acetate stem bark extract of B. coriacea can be attributed to the presence of bioactive phytochemicals, such as phenolics, flavonoids, alkaloids, some types of phytosterols and terpenoids. These phytochemicals have been shown to exert anticancer activity against various cancers through various mechanisms [34].

Selectivity index (SI) is a robust parameter to deduce the ability of plant extracts and chemicals to inhibit the proliferation of cancerous cells while selectively sparing normal/health cells [35]. In this study, the ethyl acetate stem bark extract of B. coriacea exhibited high selectivity against DU-145 cells, compared to Doxorubicin’s moderate electivity, underscoring the presence of effective phytochemicals which can target unique processes of cancerous cells to inhibit their proliferation and survival. Additionally, the presence of antioxidant compounds in this extract, certainly thwarted free radical-mediated oxidative damage to essential biological molecules in healthy mammalian cells, while perpetuating damage to cancerous cells as reported previously [36].

The primary mechanism of action attributed to doxorubicin involves its inherent capability to intercalate within DNA pairs, thereby instigating the fragmentation of DNA strands and impeding DNA and RNA synthesis [37]. Additionally, doxorubicin precipitates free radical-mediated oxidative damage to DNA in conjunction with iron, imposing supplementary restrictions on DNA synthesis [38]. However, the lower selectivity of doxorubicin against cancerous cells is associated with adverse effects, such as fatigue, alopecia, nausea, vomiting, and oral sores, along with notable instances of bone marrow suppression and elevated susceptibility to secondary malignancies, among other devastating complications [38, 39]. Thus, considering the drawbacks bedevilling doxorubicin therapy and other conventional chemotherapeutic agents, the utilization of the promising plant extracts and their valorisation based on empirical evidence, such as this study, is a viable alternative to mitigating the adverse effects. Notably, the high selectivity of the ethyl acetate stem bark extract of B. coriacea can be linked to the presence of phytochemicals which induced DNA and RNA damage in the DU-145 cells, while sparing health cells (Vero CCL-81), underpinning its anticancer efficacy and safety. However, further empirical investigations, using other cell lines and in vivo models are required to clarify the promising effects reported herein, and to facilitate the development of safe and effective botanically-derived anticancer agents.

The progression of PCa is a complex process involving the dysregulation of critical genes governing the cell cycle, proto-oncogenes, and tumour-suppressing genes [40]. The androgen receptor (AR) plays a crucial role in PCa progression [41]. When activated through its ligand binding domain (LBD), the AR translocates into the nucleus and forms dimers that bind to the androgen response element (ARE) on target genes [42], recruiting transcriptional machinery, and facilitating gene transcription [43]. Pharmacological compounds inhibiting AR action target either the ligand binding domain (LBD) of the androgen receptor gene (ar) or interactions with co-regulatory proteins, impeding disease progression [44]. It was observed that the expression of the ar gene in PCa cells (DU-145) treated with the ethyl acetate stem bark extract of B. coriacea was significantly lower than in the control cells. These findings imply that the antiproliferative effects of this extract observed in this study are partly driven through the inhibition of the ar gene overexpression [45].

The B-Cell Lymphoma-2 (BCL-2) family of proteins comprise anti-apoptotic proteins (BCL-2, BCL-XL, MCL-1, BCL-W, and BFL-1) and pro-apoptotic proteins (BAK, BAX, BOK and BH3-only proteins). The pro-apoptotic proteins are categorised as sensitizers (NOXA, BAD, HRK, BMF, and BIK) and activators (BID, BIM, and PUMA) [46]. The intricate interplay between pro- and anti-apoptotic protein levels determines a cell’s fate- whether it lives or dies [47]. The expression of the bcl-2 gene was significantly lower in DU-145 cells treated with the ethyl acetate stem bark extract of B. coriacea than in the control cells. This finding underscores the potential of the studied plant extract potential in averting the bcl-2 overexpression in PCa cells to promote their apoptosis.

The Caspase family represents a group of proteins, homologous to the Caenorhabditis elegans’ cell death abnormal-3 gene (CED-3) [48]. This protein family is distinctly composed of initiator caspases (caspase-2, -8, -9, -10), executioner caspases (caspase-3, -6, -7), and inflammatory caspases (caspase-1, -4, -5, -11) [49]. Depletion of caspase activity, especially caspase-3, plays a crucial role in tumour progression, as it occupies a terminal position in the caspase cascade, activated by both intrinsic and extrinsic death pathways in apoptosis [50]. Previous research shows that the overexpression of caspase 3 may reduce cancer patients’ overall survival rate and confer tumour cell regrowth ability, chemotherapy immunity, and other adverse effects [51]. Notably, the expression of caspase-3 in DU-145 cells treated with the studied plant extract was normal, denoting the extract’s ability to normalise its expression level to promote the execution of apoptosis [50].

Cyclin-dependent kinases (CDKs) are serine/threonine kinases that form complexes with cyclin proteins, a crucial process for fully activating their kinase activity [52]. Their pivotal roles extend to controlling cell division and transcription modulation in response to extracellular and intracellular stimuli [53]. Analysis from The Cancer Genome Atlas (TCGA) UALCAN database reveals significant upregulation of CDKs in cancerous tissues compared to normal tissues [54]. This study’s findings depicted a normal expression of CDK1 in DU-145 cells treated with the ethyl acetate stem bark extract of B. coriacea. These results underscore this extract’s ability to restore normal expression of CDK1 and probably its normal function in regulating the cell cycle, thus averting PCa. Further investigation of this extract’s modulatory role on the cell cycle using other in vitro and in vivo models may provide crucial insights into its anticancer potential.

The p53 gene encodes the p53 tumour suppressor protein, commonly known as the "Guardian of the Genome," which safeguards the integrity of cellular DNA and plays pivotal roles in development, ageing, and cell differentiation [55]. Genetic models with p53-nullizygous traits display phenotypes related to ageing, pluripotency, and development, characterised by early onset of ageing, induction of cell pluripotency, and the inability of embryos to undergo gastrulation [56]. Recent studies have revealed p53’s involvement in pathways such as autophagy, cell metabolism, ferroptosis, and those generating reactive oxygen species [56]. The significantly higher expression of p53 in DU-145 cells treated with the ethyl acetate stem bark extract of B. coriacea indicates the presence of key secondary metabolites which not only reactivate the expression of this gene but also increase its expression levels to promote apoptosis in prostate cancer cells [57]. Furthermore, determining the intricate mechanisms through which the studied plant extract influence the interplay of other genes may unravel the appropriate targets and facilitate the development of efficacious and safe anti-PCa drugs from this botanical resource.

Phytochemical analysis of the ethyl acetate stem bark extract of B. coriacea revealed a wide range of compounds, including triterpenoids, phenolics, and fatty acid esters. For instance, triterpenoids like lup-20(29)-en-3-one and lupeol, detected in this extract possess various biological effects, including anti-inflammatory, antioxidant, and anticancer activities [58]. Existing evidence shows these compounds prevent the development of cancer, by promoting apoptosis and inhibiting cancer cell proliferation and metastasis [58]. Lupeol exerts its anticancer effects by modulating key signalling pathways such as PI3K/Akt, MAPK/ERK, and JAK/STAT, and regulating the function of proteins driving cell cycle progression or apoptosis [58]. Besides, the high antioxidant and antiproliferative effects of the studied plant extract can be attributed to the high concentration of these phytochemicals. Research has demonstrated that phytosterols promote proper immune response functioning while also improving gut health by modulating gut microbiome [59]. Therefore, the detection of these phytosterols in the studied plant extract underpins its pharmacological value, yet to be fully exploited. The detection of neophytadiene, 1-Heneicosanol, γ-Sitosterol.

Lup-20(29)-en-3-one, and Lupeol implies that the extract possesses anti-inflammatory and antioxidant properties, which, in turn may contribute to its anti-cancer effects [60].

Therefore, considering the ethnomedicinal utilisation of the studied plant in treating PCa and other OS-associated diseases [10], this initial report partly validates its efficacy, and pave the way for further studies that may lead to the isolation and characterisation of lead compounds for the development of safe and potent armamentaria against PCa as well as other OS-related diseases. Considering the preliminary nature of this study, additional investigations using other in vitro and in vivo models are required to comprehensively appraise this extract’s safety and therapeutic potential. Moreover, such investigations will provide crucial empirical data that may guide the translation of research findings into clinical practice.

5. Conclusions and recommendations

The ethyl acetate stem bark extract of B. coriacea exhibited promising in vitro antioxidant and anti-prostate cancer properties. The observed anti-prostate cancer effects are likely mediated through the modulation of key molecular pathways, including the downregulation of androgen receptor (ar) and B-cell lymphoma 2 (bcl-2), normalization of caspase-3 and cyclin-dependent kinase 1 (cdk1), and upregulation of p53, ultimately inducing apoptosis. The presence of bioactive phytochemicals in the studied plant extract further underscores its therapeutic potential, and call for more comprehensive investigations using a variety of cancer cell lines and in vivo models to facilitate the isolation and empirical characterisation of lead compounds for the development of targeted anticancer therapies. Given its high selectivity toward prostate cancer cells and favorable safety profile, the ethyl acetate stem bark extract of B. coriacea is a promising candidate for further pharmacological exploration to unlock its full therapeutic potential.

Supporting information

S1 File. Descriptive and inferential statistical data of the DPPH radical scavenging activity ethyl acetate stem bark extract of B. coriacea.

https://doi.org/10.1371/journal.pone.0309258.s001

(XLSX)

S2 File. Descriptive and inferential statistical data of the hydrogen peroxide scavenging activity of ethyl acetate stem bark extract of B. coriacea.

https://doi.org/10.1371/journal.pone.0309258.s002

(XLSX)

S3 File. Descriptive and inferential statistical data of the cytotoxicity of ethyl acetate stem bark extract of B. coriacea against Vero CCL-81 cells.

https://doi.org/10.1371/journal.pone.0309258.s003

(XLSX)

S4 File. Descriptive and inferential statistical data of the inhibitory effects of the ethyl acetate stem bark extract of B. coriacea against prostate cancer cells (DU-145).

https://doi.org/10.1371/journal.pone.0309258.s004

(XLSX)

Acknowledgments

We are grateful for the technical assistance provided by the laboratory technologists, particularly Mr. Daniel Mwaniki (Department of Biochemistry, Microbiology, and Biotechnology, Kenyatta University), Mr. Elias Mandela (Department of Biological Sciences, Mount Kenya University), and Ms. Sally Kamau and Mr. Gilbert Kipkorir (Centre for Traditional Medicine and Drug Research-Kenya Medical Research Institute).

References

1. Forman HJ, Zhang H. Targeting oxidative stress in disease: promise and limitations of antioxidant therapy. Nature Reviews Drug Discovery. Nature Research; 2021. pp. 689–709. pmid:34194012
- View Article
- PubMed/NCBI
- Google Scholar
2. Liguori I, Russo G, Curcio F, Bulli G, Aran L, Della-Morte D, et al. Oxidative stress, aging, and diseases. Clin Interv Aging. 2018;13: 757–772. pmid:29731617
- View Article
- PubMed/NCBI
- Google Scholar
3. Pires MA, Munekata PES, Villanueva NDM, Tonin FG, Baldin JC, Rocha YJP, et al. The antioxidant capacity of rosemary and green tea extracts to replace the carcinogenic antioxidant (BHA) in chicken burgers. J Food Qual. 2017;2017.
- View Article
- Google Scholar
4. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71: 209–249. pmid:33538338
- View Article
- PubMed/NCBI
- Google Scholar
5. Zi H, He SH, Leng XY, Xu XF, Huang Q, Weng H, et al. Global, regional, and national burden of kidney, bladder, and prostate cancers and their attributable risk factors, 1990–2019. Mil Med Res. 2021;8. pmid:34819142
- View Article
- PubMed/NCBI
- Google Scholar
6. Ministry of Health (MOH)-Kenya. National Strategic Plan For The Prevention And Control Of Non-Communicable Diseases (NCDs) 2021/22-2025/26. Nairobi; 2021. www.health.go.ke
7. Lowder D, Rizwan K, McColl C, Paparella A, Ittmann M, Mitsiades N, et al. Racial disparities in prostate cancer: A complex interplay between socioeconomic inequities and genomics. Cancer Lett. 2022;531: 71–82. pmid:35122875
- View Article
- PubMed/NCBI
- Google Scholar
8. Ocran Mattila P, Ahmad R, Hasan SS, Babar ZUD. Availability, Affordability, Access, and Pricing of Anti-cancer Medicines in Low- and Middle-Income Countries: A Systematic Review of Literature. Frontiers in Public Health. Frontiers Media S.A.; 2021. pmid:33996712
- View Article
- PubMed/NCBI
- Google Scholar
9. Steentjes L, Siesling S, Drummond FJ, van Manen JG, Sharp L, Gavin A. Factors associated with current and severe physical side-effects after prostate cancer treatment: What men report. Eur J Cancer Care (Engl). 2018;27. pmid:27726215
- View Article
- PubMed/NCBI
- Google Scholar
10. Maroyi A. Boscia coriacea, B. foetida and B. mossambicensis: comparative analysis of their medicinal uses and ethnopharmacological properties. J Pharm Sci & Res. 2019;11: 3116–3121.
- View Article
- Google Scholar
11. Carter R, Bryson CT, Darbyshire SJ. Preparation and Use of Voucher Specimens for Documenting Research in Weed Science. Weed Technology. 2007;21: 1101–1108.
- View Article
- Google Scholar
12. Harborne JB. Phytochemical Methods A Guide To Modern Tecniques Of Plant Analysis, Third Edition. Chapman & Hall. 1998.
13. Moriasi GA, Ireri AM, Nelson EM, Ngugi MP. In vivo anti-inflammatory, anti-nociceptive, and in vitro antioxidant efficacy, and acute oral toxicity effects of the aqueous and methanolic stem bark extracts of Lonchocarpus eriocalyx (Harms.). Heliyon. 2021;7: e07145. pmid:34136700
- View Article
- PubMed/NCBI
- Google Scholar
14. Truong D-H, Nguyen DH, Ta NTA, Bui AV, Do TH, Nguyen HC. Evaluation of the Use of Different Solvents for Phytochemical Constituents, Antioxidants, and In Vitro Anti-Inflammatory Activities of Severinia buxifolia. J Food Qual. 2019;2019: 1–9.
- View Article
- Google Scholar
15. Brand-Williams W; Cuvelier ME, Berset C. Use of a Free Radical Method to Evaluate Antioxidant Activity. 1995;30: 25–30.
- View Article
- Google Scholar
16. Kibiti CM, Afolayan AJ. Preliminary Phytochemical Screening and Biological Activities of Bulbine abyssinica Used in the Folk Medicine in the Eastern Cape Province, South Africa. Evidence-based Complementary and Alternative Medicine. 2015;2015. pmid:26579202
- View Article
- PubMed/NCBI
- Google Scholar
17. Aliyu Abubakar B., Ibrahim Mohammed A., Musa Aliyu M., Musa Aisha O., Kiplimo Joyce J., Oyewale Adebayo O. Free Radical Scavenging And Total Antioxidant Capacity Of Root Extracts Of Anchomanes difformis Engl. (Araceae). Acta Poloniae Pharmaceutica ñ Drug Research. 2013;70: 115–121. pmid:23610966
- View Article
- PubMed/NCBI
- Google Scholar
18. Markossian S, Grossman A, Brimacombe Michelle Arkin K, Auld D, Austin Jonathan Baell CP, Chung TD, et al. Assay Guidance Manual. September 1. Eli Lilly & Company and the National Center for Advancing Translational Sciences Bethesda (MD); 2021. https://ncats.nih.gov/expertise/preclinical/agm.
19. van Meerloo J, Kaspers GJL, Cloos J. Cell sensitivity assays: The MTT assay. Methods in Molecular Biology. 2011;731: 237–245. pmid:21516412
- View Article
- PubMed/NCBI
- Google Scholar
20. Siti Syarifah MM, Nurhanan M, Muhd Haffiz J, Mohd Ilham A, Getha K, Asiah O, et al. Potential anticancer compound from Cerbera odollam. Journal of Tropical Forest Science. 2011;23: 89–96.
- View Article
- Google Scholar
21. Rao X, Huang X, Zhou Z, Lin X. An improvement of the 2ˆ(-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 2013;3: 71–85.
- View Article
- Google Scholar
22. Nagaraja SK, Nayaka S, Kumar RS. Phytochemical Analysis, GC–MS Profiling, and In Vitro Evaluation of Biological Applications of Different Solvent Extracts of Leonotis nepetifolia (L.) R.Br. Flower Buds. Appl Biochem Biotechnol. 2023;195: 1197–1215. pmid:36342622
- View Article
- PubMed/NCBI
- Google Scholar
23. Eseberri I, Trepiana J, Léniz A, Gómez-García I, Carr-Ugarte H, González M, et al. Variability in the Beneficial Effects of Phenolic Compounds: A Review. Nutrients. MDPI; 2022. pmid:35565892
- View Article
- PubMed/NCBI
- Google Scholar
24. Moriasi G, Ireri A, Ngugi MP. In vitro antioxidant activities of the aqueous and methanolic stem bark extracts of Piliostigma thonningii (Schum.). J Evid Based Integr Med. 2020;25: 2515690X20937988. pmid:32664742
- View Article
- PubMed/NCBI
- Google Scholar
25. Fidrianny I, Budiana W, Ruslan K. Antioxidant activities of various extracts from ardisia sp leaves using dpph and cuprac assays and correlation with total flavonoid, phenolic, carotenoid content. International Journal of Pharmacognosy and Phytochemical Research. 2015;7: 859–865.
- View Article
- Google Scholar
26. Gęgotek A, Skrzydlewska E. Antioxidative and Anti-Inflammatory Activity of Ascorbic Acid. Antioxidants. MDPI; 2022. pmid:36290716
- View Article
- PubMed/NCBI
- Google Scholar
27. Ilhami Gülçin, Haci Ahmet Alici, band Mehmet Cesur. Determination of in Vitro Antioxidant and Radical Scavenging Activities of Propofol. Chem Pharm Bull (Tokyo). 2005;53: 281–285.
- View Article
- Google Scholar
28. Lipinski B. Hydroxyl radical and its scavengers in health and disease. Oxidative Medicine and Cellular Longevity. 2011. pmid:21904647
- View Article
- PubMed/NCBI
- Google Scholar
29. Gupta S, Finelli R, Agarwal A, Henkel R. Total antioxidant capacity—Relevance, methods and clinical implications. Andrologia. Blackwell Publishing Ltd; 2021. pmid:32400041
- View Article
- PubMed/NCBI
- Google Scholar
30. Ghasemi M, Turnbull T, Sebastian S, Kempson I. The mtt assay: Utility, limitations, pitfalls, and interpretation in bulk and single-cell analysis. Int J Mol Sci. 2021;22. pmid:34884632
- View Article
- PubMed/NCBI
- Google Scholar
31. Indrayanto G, Putra GS, Suhud F. Validation of in-vitro bioassay methods: Application in herbal drug research. Profiles of Drug Substances, Excipients and Related Methodology. Academic Press Inc.; 2021. pp. 273–307. pmid:33461699
- View Article
- PubMed/NCBI
- Google Scholar
32. Muruthi CW, Ngugi MP, Runo SM, Mwitari PG. In Vitro Antiproliferative Effects and Phytochemical Characterization of Carissa edulis ((Forssk) Vahl) and Pappea capensis (Eckyl and Zeyh) Extracts. J Evid Based Integr Med. 2023;28. pmid:37489007
- View Article
- PubMed/NCBI
- Google Scholar
33. Ghout A, Zellagui A, Gherraf N, Demirtas I, Ayse Sahin Y, Boukhenaf M, et al. Antiproliferative and Antioxidant Activities of Two Extracts of the Plant Species Euphorbia dendroides L. Medicines. 2018;5: 36. pmid:29677109
- View Article
- PubMed/NCBI
- Google Scholar
34. Siddiqui AJ, Jahan S, Singh R, Saxena J, Ashraf SA, Khan A, et al. Plants in Anticancer Drug Discovery: From Molecular Mechanism to Chemoprevention. BioMed Research International. Hindawi Limited; 2022.
35. Radha Abbas Hasoon M, Jawad Kadhim N. Improvement of the Selectivity Index (SI) and cytotoxicity activity of doxorubicin drug by panax ginseng plant extract. Arch Razi Inst. 2021;76: 659–666. pmid:34824758
- View Article
- PubMed/NCBI
- Google Scholar
36. Raletsena M V., Mongalo NI. Phytochemical analysis, in vitro antimicrobial, anticancer, anti-inflammatory, and antioxidant activity of extracts from Bulbine anguistifolia Poelln (Asphodelaceae). South African Journal of Botany. 2023;159: 588–595.
- View Article
- Google Scholar
37. Thorn CF, Oshiro C, Marsh S, Hernandez-Boussard T, McLeod H, Klein TE, et al. Doxorubicin pathways: Pharmacodynamics and adverse effects. Pharmacogenet Genomics. 2011;21: 440–446. pmid:21048526
- View Article
- PubMed/NCBI
- Google Scholar
38. Sritharan S, Sivalingam N. A comprehensive review on time-tested anticancer drug doxorubicin. Life Sciences. Elsevier Inc.; 2021. pmid:33887349
- View Article
- PubMed/NCBI
- Google Scholar
39. Lai C, Cole DE, Steinberg SM, Lucas N, Dombi E, Melani C, et al. Doxorubicin pharmacokinetics and toxicity in patients with aggressive lymphoma and hepatic impairment. Blood Adv. 2023;7: 529–532. pmid:35882475
- View Article
- PubMed/NCBI
- Google Scholar
40. Somarelli JA, Gardner H, Cannataro VL, Gunady EF, Boddy AM, Johnson NA, et al. Molecular Biology and Evolution of Cancer: From Discovery to Action. Molecular Biology and Evolution. Oxford University Press; 2020. pp. 320–326. pmid:31642480
- View Article
- PubMed/NCBI
- Google Scholar
41. Eisermann K, Fraizer G. The androgen receptor and VEGF: Mechanisms of androgen-regulated angiogenesis in prostate cancer. Cancers. MDPI AG; 2017. pmid:28394264
- View Article
- PubMed/NCBI
- Google Scholar
42. Cheng Y, Yu P, Duan X, Liu C, Xu S, Chen Y, et al. Genome-wide analysis of androgen receptor binding sites in prostate cancer cells. Exp Ther Med. 2015;9: 2319–2324. pmid:26136980
- View Article
- PubMed/NCBI
- Google Scholar
43. Fujita K, Nonomura N. Role of androgen receptor in prostate cancer: A review. World Journal of Men?s Health. Korean Society for Sexual Medicine and Andrology; 2019. pp. 288–295. pmid:30209899
- View Article
- PubMed/NCBI
- Google Scholar
44. Shah K, Gagliano T, Garland L, O’Hanlon T, Bortolotti D, Gentili V, et al. Androgen receptor signaling regulates the transcriptome of prostate cancer cells by modulating global alternative splicing. Oncogene. 2020;39: 6172–6189. pmid:32820253
- View Article
- PubMed/NCBI
- Google Scholar
45. Roszkowska KA, Piecuch A, Sady M, Gajewski Z, Flis S. Gain of Function (GOF) Mutant p53 in Cancer-Current Therapeutic Approaches. Int J Mol Sci. 2022;2022: 23. pmid:36362074
- View Article
- PubMed/NCBI
- Google Scholar
46. Warren CFA, Wong-Brown MW, Bowden NA. BCL-2 family isoforms in apoptosis and cancer. Cell Death and Disease. Nature Publishing Group; 2019. pmid:30792387
- View Article
- PubMed/NCBI
- Google Scholar
47. Alam M, Ali S, Mohammad T, Hasan GM, Yadav DK, Hassan MI. B cell lymphoma 2: A potential therapeutic target for cancer therapy. International Journal of Molecular Sciences. MDPI; 2021. pmid:34638779
- View Article
- PubMed/NCBI
- Google Scholar
48. Olsson M, Zhivotovsky B. Caspases and cancer. Cell Death and Differentiation. 2011. pp. 1441–1449. pmid:21455218
- View Article
- PubMed/NCBI
- Google Scholar
49. Shalini S, Dorstyn L, Dawar S, Kumar S. Old, new and emerging functions of caspases. Cell Death and Differentiation. Nature Publishing Group; 2015. pp. 526–539. pmid:25526085
- View Article
- PubMed/NCBI
- Google Scholar
50. Eskandari E, Eaves CJ. Paradoxical roles of caspase-3 in regulating cell survival, proliferation, and tumorigenesis. Journal of Cell Biology. Rockefeller University Press; 2022. pmid:35551578
- View Article
- PubMed/NCBI
- Google Scholar
51. Yang X, Zhong D-N, Qin H, Wu P-R, Wei K-L, Chen G, et al. Caspase-3 over-expression is associated with poor overall survival and clinicopathological parameters in breast cancer: a meta-analysis of 3091 cases. Oncotarget. 2018. www.impactjournals.com/oncotarget/ pmid:29492222
- View Article
- PubMed/NCBI
- Google Scholar
52. Malumbres M. Cyclin-dependent kinases. Genome Biol. 2014;15. pmid:25180339
- View Article
- PubMed/NCBI
- Google Scholar
53. Łukasik P, Załuski M, Gutowska I. Cyclin-dependent kinases (Cdk) and their role in diseases development–review. International Journal of Molecular Sciences. MDPI AG; 2021. pp. 1–33. pmid:33805800
- View Article
- PubMed/NCBI
- Google Scholar
54. Chandrashekar DS, Karthikeyan SK, Korla PK, Patel H, Shovon AR, Athar M, et al. UALCAN: An update to the integrated cancer data analysis platform. Neoplasia (United States). 2022;25: 18–27. pmid:35078134
- View Article
- PubMed/NCBI
- Google Scholar
55. Huang H, Tang Y, Li P, Ye X, Chen W, Xie H, et al. Significance of TP53 and immune-related genes to prostate cancer. Transl Androl Urol. 2021;10: 1754–1768. pmid:33968663
- View Article
- PubMed/NCBI
- Google Scholar
56. Hernández Borrero LJ, El-Deiry WS. Tumor suppressor p53: Biology, signaling pathways, and therapeutic targeting. Biochimica et Biophysica Acta—Reviews on Cancer. Elsevier B.V.; 2021. pmid:33932560
- View Article
- PubMed/NCBI
- Google Scholar
57. Wang H, Guo M, Wei H, Chen Y. Targeting p53 pathways: mechanisms, structures, and advances in therapy. Signal Transduction and Targeted Therapy. Springer Nature; 2023. pmid:36859359
- View Article
- PubMed/NCBI
- Google Scholar
58. Liu K, Zhang X, Xie L, Deng M, Chen H, Song J, et al. Lupeol and its derivatives as anticancer and anti-inflammatory agents: Molecular mechanisms and therapeutic efficacy. Pharmacological Research. Academic Press; 2021. pmid:33316380
- View Article
- PubMed/NCBI
- Google Scholar
59. Li B, Yang Z, Mao F, Wang Q, Fang H, Gu X, et al. Phytochemical profile and biological activities of the essential oils in the aerial part and root of Saposhnikovia divaricata. Sci Rep. 2023;13. pmid:37248268
- View Article
- PubMed/NCBI
- Google Scholar
60. Zhang X, Wang J, Zhu L, Wang X, Meng F, Xia L, et al. Advances in Stigmasterol on its anti-tumor effect and mechanism of action. Frontiers in Oncology. Frontiers Media S.A.; 2022. pmid:36578938
- View Article
- PubMed/NCBI
- Google Scholar

[ref1] 1. Forman HJ, Zhang H. Targeting oxidative stress in disease: promise and limitations of antioxidant therapy. Nature Reviews Drug Discovery. Nature Research; 2021. pp. 689–709. pmid:34194012
View Article
PubMed/NCBI
Google Scholar

[2] View Article

[3] PubMed/NCBI

[4] Google Scholar

[ref2] 2. Liguori I, Russo G, Curcio F, Bulli G, Aran L, Della-Morte D, et al. Oxidative stress, aging, and diseases. Clin Interv Aging. 2018;13: 757–772. pmid:29731617
View Article
PubMed/NCBI
Google Scholar

[6] View Article

[7] PubMed/NCBI

[8] Google Scholar

[ref3] 3. Pires MA, Munekata PES, Villanueva NDM, Tonin FG, Baldin JC, Rocha YJP, et al. The antioxidant capacity of rosemary and green tea extracts to replace the carcinogenic antioxidant (BHA) in chicken burgers. J Food Qual. 2017;2017.
View Article
Google Scholar

[10] View Article

[11] Google Scholar

[ref4] 4. Sung H, Ferlay J, Siegel RL, Laversanne M, Soerjomataram I, Jemal A, et al. Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries. CA Cancer J Clin. 2021;71: 209–249. pmid:33538338
View Article
PubMed/NCBI
Google Scholar

[13] View Article

[14] PubMed/NCBI

[15] Google Scholar

[ref5] 5. Zi H, He SH, Leng XY, Xu XF, Huang Q, Weng H, et al. Global, regional, and national burden of kidney, bladder, and prostate cancers and their attributable risk factors, 1990–2019. Mil Med Res. 2021;8. pmid:34819142
View Article
PubMed/NCBI
Google Scholar

[17] View Article

[18] PubMed/NCBI

[19] Google Scholar

[ref6] 6. Ministry of Health (MOH)-Kenya. National Strategic Plan For The Prevention And Control Of Non-Communicable Diseases (NCDs) 2021/22-2025/26. Nairobi; 2021. www.health.go.ke

[ref7] 7. Lowder D, Rizwan K, McColl C, Paparella A, Ittmann M, Mitsiades N, et al. Racial disparities in prostate cancer: A complex interplay between socioeconomic inequities and genomics. Cancer Lett. 2022;531: 71–82. pmid:35122875
View Article
PubMed/NCBI
Google Scholar

[22] View Article

[23] PubMed/NCBI

[24] Google Scholar

[ref8] 8. Ocran Mattila P, Ahmad R, Hasan SS, Babar ZUD. Availability, Affordability, Access, and Pricing of Anti-cancer Medicines in Low- and Middle-Income Countries: A Systematic Review of Literature. Frontiers in Public Health. Frontiers Media S.A.; 2021. pmid:33996712
View Article
PubMed/NCBI
Google Scholar

[26] View Article

[27] PubMed/NCBI

[28] Google Scholar

[ref9] 9. Steentjes L, Siesling S, Drummond FJ, van Manen JG, Sharp L, Gavin A. Factors associated with current and severe physical side-effects after prostate cancer treatment: What men report. Eur J Cancer Care (Engl). 2018;27. pmid:27726215
View Article
PubMed/NCBI
Google Scholar

[30] View Article

[31] PubMed/NCBI

[32] Google Scholar

[ref10] 10. Maroyi A. Boscia coriacea, B. foetida and B. mossambicensis: comparative analysis of their medicinal uses and ethnopharmacological properties. J Pharm Sci & Res. 2019;11: 3116–3121.
View Article
Google Scholar

[34] View Article

[35] Google Scholar

[ref11] 11. Carter R, Bryson CT, Darbyshire SJ. Preparation and Use of Voucher Specimens for Documenting Research in Weed Science. Weed Technology. 2007;21: 1101–1108.
View Article
Google Scholar

[37] View Article

[38] Google Scholar

[ref12] 12. Harborne JB. Phytochemical Methods A Guide To Modern Tecniques Of Plant Analysis, Third Edition. Chapman & Hall. 1998.

[ref13] 13. Moriasi GA, Ireri AM, Nelson EM, Ngugi MP. In vivo anti-inflammatory, anti-nociceptive, and in vitro antioxidant efficacy, and acute oral toxicity effects of the aqueous and methanolic stem bark extracts of Lonchocarpus eriocalyx (Harms.). Heliyon. 2021;7: e07145. pmid:34136700
View Article
PubMed/NCBI
Google Scholar

[41] View Article

[42] PubMed/NCBI

[43] Google Scholar

[ref14] 14. Truong D-H, Nguyen DH, Ta NTA, Bui AV, Do TH, Nguyen HC. Evaluation of the Use of Different Solvents for Phytochemical Constituents, Antioxidants, and In Vitro Anti-Inflammatory Activities of Severinia buxifolia. J Food Qual. 2019;2019: 1–9.
View Article
Google Scholar

[45] View Article

[46] Google Scholar

[ref15] 15. Brand-Williams W; Cuvelier ME, Berset C. Use of a Free Radical Method to Evaluate Antioxidant Activity. 1995;30: 25–30.
View Article
Google Scholar

[48] View Article

[49] Google Scholar

[ref16] 16. Kibiti CM, Afolayan AJ. Preliminary Phytochemical Screening and Biological Activities of Bulbine abyssinica Used in the Folk Medicine in the Eastern Cape Province, South Africa. Evidence-based Complementary and Alternative Medicine. 2015;2015. pmid:26579202
View Article
PubMed/NCBI
Google Scholar

[51] View Article

[52] PubMed/NCBI

[53] Google Scholar

[ref17] 17. Aliyu Abubakar B., Ibrahim Mohammed A., Musa Aliyu M., Musa Aisha O., Kiplimo Joyce J., Oyewale Adebayo O. Free Radical Scavenging And Total Antioxidant Capacity Of Root Extracts Of Anchomanes difformis Engl. (Araceae). Acta Poloniae Pharmaceutica ñ Drug Research. 2013;70: 115–121. pmid:23610966
View Article
PubMed/NCBI
Google Scholar

[55] View Article

[56] PubMed/NCBI

[57] Google Scholar

[ref18] 18. Markossian S, Grossman A, Brimacombe Michelle Arkin K, Auld D, Austin Jonathan Baell CP, Chung TD, et al. Assay Guidance Manual. September 1. Eli Lilly & Company and the National Center for Advancing Translational Sciences Bethesda (MD); 2021. https://ncats.nih.gov/expertise/preclinical/agm.

[ref19] 19. van Meerloo J, Kaspers GJL, Cloos J. Cell sensitivity assays: The MTT assay. Methods in Molecular Biology. 2011;731: 237–245. pmid:21516412
View Article
PubMed/NCBI
Google Scholar

[60] View Article

[61] PubMed/NCBI

[62] Google Scholar

[ref20] 20. Siti Syarifah MM, Nurhanan M, Muhd Haffiz J, Mohd Ilham A, Getha K, Asiah O, et al. Potential anticancer compound from Cerbera odollam. Journal of Tropical Forest Science. 2011;23: 89–96.
View Article
Google Scholar

[64] View Article

[65] Google Scholar

[ref21] 21. Rao X, Huang X, Zhou Z, Lin X. An improvement of the 2ˆ(-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 2013;3: 71–85.
View Article
Google Scholar

[67] View Article

[68] Google Scholar

[ref22] 22. Nagaraja SK, Nayaka S, Kumar RS. Phytochemical Analysis, GC–MS Profiling, and In Vitro Evaluation of Biological Applications of Different Solvent Extracts of Leonotis nepetifolia (L.) R.Br. Flower Buds. Appl Biochem Biotechnol. 2023;195: 1197–1215. pmid:36342622
View Article
PubMed/NCBI
Google Scholar

[70] View Article

[71] PubMed/NCBI

[72] Google Scholar

[ref23] 23. Eseberri I, Trepiana J, Léniz A, Gómez-García I, Carr-Ugarte H, González M, et al. Variability in the Beneficial Effects of Phenolic Compounds: A Review. Nutrients. MDPI; 2022. pmid:35565892
View Article
PubMed/NCBI
Google Scholar

[74] View Article

[75] PubMed/NCBI

[76] Google Scholar

[ref24] 24. Moriasi G, Ireri A, Ngugi MP. In vitro antioxidant activities of the aqueous and methanolic stem bark extracts of Piliostigma thonningii (Schum.). J Evid Based Integr Med. 2020;25: 2515690X20937988. pmid:32664742
View Article
PubMed/NCBI
Google Scholar

[78] View Article

[79] PubMed/NCBI

[80] Google Scholar

[ref25] 25. Fidrianny I, Budiana W, Ruslan K. Antioxidant activities of various extracts from ardisia sp leaves using dpph and cuprac assays and correlation with total flavonoid, phenolic, carotenoid content. International Journal of Pharmacognosy and Phytochemical Research. 2015;7: 859–865.
View Article
Google Scholar

[82] View Article

[83] Google Scholar

[ref26] 26. Gęgotek A, Skrzydlewska E. Antioxidative and Anti-Inflammatory Activity of Ascorbic Acid. Antioxidants. MDPI; 2022. pmid:36290716
View Article
PubMed/NCBI
Google Scholar

[85] View Article

[86] PubMed/NCBI

[87] Google Scholar

[ref27] 27. Ilhami Gülçin, Haci Ahmet Alici, band Mehmet Cesur. Determination of in Vitro Antioxidant and Radical Scavenging Activities of Propofol. Chem Pharm Bull (Tokyo). 2005;53: 281–285.
View Article
Google Scholar

[89] View Article

[90] Google Scholar

[ref28] 28. Lipinski B. Hydroxyl radical and its scavengers in health and disease. Oxidative Medicine and Cellular Longevity. 2011. pmid:21904647
View Article
PubMed/NCBI
Google Scholar

[92] View Article

[93] PubMed/NCBI

[94] Google Scholar

[ref29] 29. Gupta S, Finelli R, Agarwal A, Henkel R. Total antioxidant capacity—Relevance, methods and clinical implications. Andrologia. Blackwell Publishing Ltd; 2021. pmid:32400041
View Article
PubMed/NCBI
Google Scholar

[96] View Article

[97] PubMed/NCBI

[98] Google Scholar

[ref30] 30. Ghasemi M, Turnbull T, Sebastian S, Kempson I. The mtt assay: Utility, limitations, pitfalls, and interpretation in bulk and single-cell analysis. Int J Mol Sci. 2021;22. pmid:34884632
View Article
PubMed/NCBI
Google Scholar

[100] View Article

[101] PubMed/NCBI

[102] Google Scholar

[ref31] 31. Indrayanto G, Putra GS, Suhud F. Validation of in-vitro bioassay methods: Application in herbal drug research. Profiles of Drug Substances, Excipients and Related Methodology. Academic Press Inc.; 2021. pp. 273–307. pmid:33461699
View Article
PubMed/NCBI
Google Scholar

[104] View Article

[105] PubMed/NCBI

[106] Google Scholar

[ref32] 32. Muruthi CW, Ngugi MP, Runo SM, Mwitari PG. In Vitro Antiproliferative Effects and Phytochemical Characterization of Carissa edulis ((Forssk) Vahl) and Pappea capensis (Eckyl and Zeyh) Extracts. J Evid Based Integr Med. 2023;28. pmid:37489007
View Article
PubMed/NCBI
Google Scholar

[108] View Article

[109] PubMed/NCBI

[110] Google Scholar

[ref33] 33. Ghout A, Zellagui A, Gherraf N, Demirtas I, Ayse Sahin Y, Boukhenaf M, et al. Antiproliferative and Antioxidant Activities of Two Extracts of the Plant Species Euphorbia dendroides L. Medicines. 2018;5: 36. pmid:29677109
View Article
PubMed/NCBI
Google Scholar

[112] View Article

[113] PubMed/NCBI

[114] Google Scholar

[ref34] 34. Siddiqui AJ, Jahan S, Singh R, Saxena J, Ashraf SA, Khan A, et al. Plants in Anticancer Drug Discovery: From Molecular Mechanism to Chemoprevention. BioMed Research International. Hindawi Limited; 2022.

[ref35] 35. Radha Abbas Hasoon M, Jawad Kadhim N. Improvement of the Selectivity Index (SI) and cytotoxicity activity of doxorubicin drug by panax ginseng plant extract. Arch Razi Inst. 2021;76: 659–666. pmid:34824758
View Article
PubMed/NCBI
Google Scholar

[117] View Article

[118] PubMed/NCBI

[119] Google Scholar

[ref36] 36. Raletsena M V., Mongalo NI. Phytochemical analysis, in vitro antimicrobial, anticancer, anti-inflammatory, and antioxidant activity of extracts from Bulbine anguistifolia Poelln (Asphodelaceae). South African Journal of Botany. 2023;159: 588–595.
View Article
Google Scholar

[121] View Article

[122] Google Scholar

[ref37] 37. Thorn CF, Oshiro C, Marsh S, Hernandez-Boussard T, McLeod H, Klein TE, et al. Doxorubicin pathways: Pharmacodynamics and adverse effects. Pharmacogenet Genomics. 2011;21: 440–446. pmid:21048526
View Article
PubMed/NCBI
Google Scholar

[124] View Article

[125] PubMed/NCBI

[126] Google Scholar

[ref38] 38. Sritharan S, Sivalingam N. A comprehensive review on time-tested anticancer drug doxorubicin. Life Sciences. Elsevier Inc.; 2021. pmid:33887349
View Article
PubMed/NCBI
Google Scholar

[128] View Article

[129] PubMed/NCBI

[130] Google Scholar

[ref39] 39. Lai C, Cole DE, Steinberg SM, Lucas N, Dombi E, Melani C, et al. Doxorubicin pharmacokinetics and toxicity in patients with aggressive lymphoma and hepatic impairment. Blood Adv. 2023;7: 529–532. pmid:35882475
View Article
PubMed/NCBI
Google Scholar

[132] View Article

[133] PubMed/NCBI

[134] Google Scholar

[ref40] 40. Somarelli JA, Gardner H, Cannataro VL, Gunady EF, Boddy AM, Johnson NA, et al. Molecular Biology and Evolution of Cancer: From Discovery to Action. Molecular Biology and Evolution. Oxford University Press; 2020. pp. 320–326. pmid:31642480
View Article
PubMed/NCBI
Google Scholar

[136] View Article

[137] PubMed/NCBI

[138] Google Scholar

[ref41] 41. Eisermann K, Fraizer G. The androgen receptor and VEGF: Mechanisms of androgen-regulated angiogenesis in prostate cancer. Cancers. MDPI AG; 2017. pmid:28394264
View Article
PubMed/NCBI
Google Scholar

[140] View Article

[141] PubMed/NCBI

[142] Google Scholar

[ref42] 42. Cheng Y, Yu P, Duan X, Liu C, Xu S, Chen Y, et al. Genome-wide analysis of androgen receptor binding sites in prostate cancer cells. Exp Ther Med. 2015;9: 2319–2324. pmid:26136980
View Article
PubMed/NCBI
Google Scholar

[144] View Article

[145] PubMed/NCBI

[146] Google Scholar

[ref43] 43. Fujita K, Nonomura N. Role of androgen receptor in prostate cancer: A review. World Journal of Men?s Health. Korean Society for Sexual Medicine and Andrology; 2019. pp. 288–295. pmid:30209899
View Article
PubMed/NCBI
Google Scholar

[148] View Article

[149] PubMed/NCBI

[150] Google Scholar

[ref44] 44. Shah K, Gagliano T, Garland L, O’Hanlon T, Bortolotti D, Gentili V, et al. Androgen receptor signaling regulates the transcriptome of prostate cancer cells by modulating global alternative splicing. Oncogene. 2020;39: 6172–6189. pmid:32820253
View Article
PubMed/NCBI
Google Scholar

[152] View Article

[153] PubMed/NCBI

[154] Google Scholar

[ref45] 45. Roszkowska KA, Piecuch A, Sady M, Gajewski Z, Flis S. Gain of Function (GOF) Mutant p53 in Cancer-Current Therapeutic Approaches. Int J Mol Sci. 2022;2022: 23. pmid:36362074
View Article
PubMed/NCBI
Google Scholar

[156] View Article

[157] PubMed/NCBI

[158] Google Scholar

[ref46] 46. Warren CFA, Wong-Brown MW, Bowden NA. BCL-2 family isoforms in apoptosis and cancer. Cell Death and Disease. Nature Publishing Group; 2019. pmid:30792387
View Article
PubMed/NCBI
Google Scholar

[160] View Article

[161] PubMed/NCBI

[162] Google Scholar

[ref47] 47. Alam M, Ali S, Mohammad T, Hasan GM, Yadav DK, Hassan MI. B cell lymphoma 2: A potential therapeutic target for cancer therapy. International Journal of Molecular Sciences. MDPI; 2021. pmid:34638779
View Article
PubMed/NCBI
Google Scholar

[164] View Article

[165] PubMed/NCBI

[166] Google Scholar

[ref48] 48. Olsson M, Zhivotovsky B. Caspases and cancer. Cell Death and Differentiation. 2011. pp. 1441–1449. pmid:21455218
View Article
PubMed/NCBI
Google Scholar

[168] View Article

[169] PubMed/NCBI

[170] Google Scholar

[ref49] 49. Shalini S, Dorstyn L, Dawar S, Kumar S. Old, new and emerging functions of caspases. Cell Death and Differentiation. Nature Publishing Group; 2015. pp. 526–539. pmid:25526085
View Article
PubMed/NCBI
Google Scholar

[172] View Article

[173] PubMed/NCBI

[174] Google Scholar

[ref50] 50. Eskandari E, Eaves CJ. Paradoxical roles of caspase-3 in regulating cell survival, proliferation, and tumorigenesis. Journal of Cell Biology. Rockefeller University Press; 2022. pmid:35551578
View Article
PubMed/NCBI
Google Scholar

[176] View Article

[177] PubMed/NCBI

[178] Google Scholar

[ref51] 51. Yang X, Zhong D-N, Qin H, Wu P-R, Wei K-L, Chen G, et al. Caspase-3 over-expression is associated with poor overall survival and clinicopathological parameters in breast cancer: a meta-analysis of 3091 cases. Oncotarget. 2018. www.impactjournals.com/oncotarget/ pmid:29492222
View Article
PubMed/NCBI
Google Scholar

[180] View Article

[181] PubMed/NCBI

[182] Google Scholar

[ref52] 52. Malumbres M. Cyclin-dependent kinases. Genome Biol. 2014;15. pmid:25180339
View Article
PubMed/NCBI
Google Scholar

[184] View Article

[185] PubMed/NCBI

[186] Google Scholar

[ref53] 53. Łukasik P, Załuski M, Gutowska I. Cyclin-dependent kinases (Cdk) and their role in diseases development–review. International Journal of Molecular Sciences. MDPI AG; 2021. pp. 1–33. pmid:33805800
View Article
PubMed/NCBI
Google Scholar

[188] View Article

[189] PubMed/NCBI

[190] Google Scholar

[ref54] 54. Chandrashekar DS, Karthikeyan SK, Korla PK, Patel H, Shovon AR, Athar M, et al. UALCAN: An update to the integrated cancer data analysis platform. Neoplasia (United States). 2022;25: 18–27. pmid:35078134
View Article
PubMed/NCBI
Google Scholar

[192] View Article

[193] PubMed/NCBI

[194] Google Scholar

[ref55] 55. Huang H, Tang Y, Li P, Ye X, Chen W, Xie H, et al. Significance of TP53 and immune-related genes to prostate cancer. Transl Androl Urol. 2021;10: 1754–1768. pmid:33968663
View Article
PubMed/NCBI
Google Scholar

[196] View Article

[197] PubMed/NCBI

[198] Google Scholar

[ref56] 56. Hernández Borrero LJ, El-Deiry WS. Tumor suppressor p53: Biology, signaling pathways, and therapeutic targeting. Biochimica et Biophysica Acta—Reviews on Cancer. Elsevier B.V.; 2021. pmid:33932560
View Article
PubMed/NCBI
Google Scholar

[200] View Article

[201] PubMed/NCBI

[202] Google Scholar

[ref57] 57. Wang H, Guo M, Wei H, Chen Y. Targeting p53 pathways: mechanisms, structures, and advances in therapy. Signal Transduction and Targeted Therapy. Springer Nature; 2023. pmid:36859359
View Article
PubMed/NCBI
Google Scholar

[204] View Article

[205] PubMed/NCBI

[206] Google Scholar

[ref58] 58. Liu K, Zhang X, Xie L, Deng M, Chen H, Song J, et al. Lupeol and its derivatives as anticancer and anti-inflammatory agents: Molecular mechanisms and therapeutic efficacy. Pharmacological Research. Academic Press; 2021. pmid:33316380
View Article
PubMed/NCBI
Google Scholar

[208] View Article

[209] PubMed/NCBI

[210] Google Scholar

[ref59] 59. Li B, Yang Z, Mao F, Wang Q, Fang H, Gu X, et al. Phytochemical profile and biological activities of the essential oils in the aerial part and root of Saposhnikovia divaricata. Sci Rep. 2023;13. pmid:37248268
View Article
PubMed/NCBI
Google Scholar

[212] View Article

[213] PubMed/NCBI

[214] Google Scholar

[ref60] 60. Zhang X, Wang J, Zhu L, Wang X, Meng F, Xia L, et al. Advances in Stigmasterol on its anti-tumor effect and mechanism of action. Frontiers in Oncology. Frontiers Media S.A.; 2022. pmid:36578938
View Article
PubMed/NCBI
Google Scholar

[216] View Article

[217] PubMed/NCBI

[218] Google Scholar

Figures

Abstract

Background

Methods

Results

Conclusion

1. Introduction

2. Materials and methods

2.1 Plant material

2.2 Extraction

2.3 Determination of in vitro antioxidant effects of the study plant extract

2.3.1 Determination of 1,1-Diphenyl-2-picryl Hydrazyl Radical (DPPH) scavenging activity.

2.3.2 Hydrogen peroxide scavenging activity.

2.3.3 Determination of total antioxidant capacity.

2.4 Determination of in vitro cytotoxic effects of the study extract against normal mammalian and PCa cells

2.4.1 Preparation of plant extracts.

2.4.2 Cell culture and maintenance.

2.4.3 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for cytotoxicity and cancer cell inhibition.

2.4.4 Determination of the expression profiles of cancer-associated genes in the DU-145 cells treated with the selected plant extract.

2.5 Phytochemical analysis

2.6 Data management, statistical analysis, and reporting

2.7 Ethics approval and consent to participate

3. Results

3.1 In vitro antioxidant activity

3.1.1 DPPH radical scavenging activity of stem bark extracts of B. coriacea.

3.1.2 Hydrogen peroxide scavenging activity of the stem bark extracts of B. coriacea.

3.1.3 Total antioxidant capacity of the stem bark extracts of B. coriacea.

3.2 In vitro cytotoxicity and antiproliferative effects

3.2.1 In vitro cytotoxicity of the ethyl acetate stem bark extract of B. coriacea.

3.2.2 Effects of the ethyl acetate stem bark extract of B. coriacea on DU-145 cells.

3.2.3 Selectivity index.

3.3 Expression levels of selected genes in DU-145 cells treated with the selected plant extracts

3.4 Phytochemical compounds detected in the ethyl acetate stem bark extract of B. coriacea

4. Discussion

5. Conclusions and recommendations

Supporting information

S1 File. Descriptive and inferential statistical data of the DPPH radical scavenging activity ethyl acetate stem bark extract of B. coriacea.

S2 File. Descriptive and inferential statistical data of the hydrogen peroxide scavenging activity of ethyl acetate stem bark extract of B. coriacea.

S3 File. Descriptive and inferential statistical data of the cytotoxicity of ethyl acetate stem bark extract of B. coriacea against Vero CCL-81 cells.

S4 File. Descriptive and inferential statistical data of the inhibitory effects of the ethyl acetate stem bark extract of B. coriacea against prostate cancer cells (DU-145).

Acknowledgments

References