Fig 1.
Assessment of Anti-Proliferative Effect of Vorinostat and ABT-199 in HCT116 and HT-29 colorectal cancer cells.
HT-29 cells and HCT-116 cells were treated with various concentrations of Vorinostat or ABT-199 for 72 hours prior to cell viability assessment by MTT assay. Cell viability percentage are presented relative to the untreated control. Data represents mean ± SD of three independent experiments.
Fig 2.
Dose-response effects and combination index (CI) analysis of Vorinostat and ABT-199 combination in colorectal cancer cells.
HT-29 (A) and HCT116 (B) cells were treated with various molar ratios of Vorinostat: ABT-199 for 72 hours, and cell viability was measured using the MTT assay. CI values for the Vorinostat and ABT-199 co-treatment were calculated in HT-29 (C) and HCT116 (D) at fraction of affected (Fa) at 0.9 (i.e., Fa of 0.9 indicates 90% cell death or growth inhibition). Data represents the mean ± SD of three independent experiments.
Table 1.
Physicochemical Characterization of BNPs, ABT-NPs, VOR-NPs and DLNPs.
Fig 3.
TEM image of DLNPs The nanoparticles appear spherical in shape with smooth surfaces and an average size of approximately 200 nm.
Scale bar = 1000 nm.
Fig 4.
Assessment of nanoparticle uptake in colorectal cancer cells using confocal microscopy and flow cytometry.
HT-29 and HCT116 cells were treated with 200 µg/mL of Rhodamine B-loaded nanoparticles for 6 hours. Following treatment, nanoparticle uptake was evaluated through flow cytometry and confocal microscopy analyses to visualize and quantify the internalization of Rhodamine B-labeled nanoparticles within the cells.
Fig 5.
Drug release kinetics of ABT-199 (A) and Vorinostat (B).
A total amount of 10 mg of each NP formulation was re-suspended in 3 ml of PBS and injected in a dialysis bag which was then immersed in a PBS containing 10% FBS. At the indicated time points, the remaining NP pellet was removed and quantified. Data represent mean ± Standard Error of the Mean (SEM).
Fig 6.
Comparative cytotoxicity of free and nanoparticle-encapsulated ABT-199 and Vorinostat in colorectal cancer cells.
(A, B) Dose–response curves of free ABT-199 versus ABT-199-loaded nanoparticles (ABT-NPs) in HT-29 and HCT116 cells, respectively, after 72-hour treatment. (C, D) Dose–response curves of free Vorinostat versus Vorinostat-loaded nanoparticles (VOR-NPs) in HT-29 and HCT116 cells, respectively. Cell viability percentage are presented relative to the untreated control. Data represents mean ± SD of three independent experiments.
Fig 7.
Apoptosis analysis of colorectal cancer cells following nanoparticle treatment using Annexin V/PI staining.
(A) HCT116 and (B) HT-29 cells were treated with BNPs, 2 µM ABT-NPs, 1 µM VOR-NPs, or equivalent concentration of DLNPs for 72 hours. Apoptosis was assessed using Annexin V-FITC/PI staining and quantified by flow cytometry. The bar graphs show the percentage of Annexin V-positive cells (early and late apoptotic), while representative dot plots depict cell distribution across quadrants (Q1 = necrotic, Q2 = late apoptotic, Q3 = early apoptotic, Q4 = viable). Data are presented as mean ± SD (n = 3). Statistical significance was determined by one-way ANOVA followed by Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.
Fig 8.
Induction of caspase-3 activity in HT-29 and HCT116 cells treated with ABT-NPs, VOR-NPs, or DLNPs. (A) HT-29 and (B) HCT116 cells were treated with BNPs, 2 µM ABT-NPs, 1 µM VOR-NPs, or equivalent concentration of DLNPs for 72 hours Caspase-3 activity was quantified using a colorimetric caspase-3 assay and expressed as fold change relative to BNPs controls.
Data are shown as mean ± SD (n = 3). Statistical significance was determined using one-way ANOVA with Tukey’s post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001.