Fig 1.
Inotodiol inhibits HCC cell viability.
(A)The influence of inotodiol on the survival of HCC cells and liver cells was examined over a 48-hour timeframe using the CCK-8 assay with a range of concentrations, n = 3. (B)The influence of inotodiol on the survival of HCC cells and liver cells was examined over a 72-hour timeframe using the CCK-8 assay with a range of concentrations, n = 3.
Fig 2.
Inotodiol inhibits sk-hep-1, hepG2 cells migration.
(A-B)Effect of inotodiol treatment of sk-hep-1 cells on the healing area of scratches and statistical results. (C-D)Effect of inotodiol treatment of hepG2 cells on the healing area of scratches and statistical results. *significantly different from control group. *p<0.05, **p<0.01, n = 3.
Fig 3.
Inotodiol decreases clone formation in HCC cells.
(A-B) The Clone formation assay and the number of clones. *significantly different from control group. ***p<0.001, ****p<0.0001, n = 3.
Fig 4.
Inotodiol-mediated cell cycle halt at the G1 phase in HCC cells.
(A-D) Inotodiol elevated the prevalence of cells within the G1 phase of the cell cycle. (E-F) Western blotting assay for the analysis of CDK2, CDK4, CDK6, cyclin D expression in inotodiol-treated sk-hep-1 cells at indicated concentration for 48h. (G-H) Western blotting assay for the analysis of CDK2, CDK4, CDK6, cyclin D expression in inotodiol-treated hepG2 cells at indicated concentration for 48h. *significantly different from control group. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, n = 3.
Fig 5.
Inotodiol triggered apoptosis in HCC cells.
(A-B) The apoptosis rate of inotodiol-treated sk-hep-1 and hepG2 cells after 48 hours was detected using flow cytometry. (C-F) Examination of the expression profiles of proteins associated with apoptosis in the sk-hep-1 (C-D) and hepG2 (E-F) cell lines was conducted using western blotting after the treatment with inotodiol.*significantly different from control group. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, n = 3.
Fig 6.
Inotodiol inhibits MAPK/ERK pathways in HCC cells.
(A-E) Protein expression and statistical results of RAS/RAF/MEK/ERK pathway related proteins in sk-hep-1 cells ascertained by Western Blot. (F-J) Protein expression and statistical results of MAPK/ERK pathway related proteins in hepG2 cells detected by Western Blot. NS, not significant different from control group, *significantly different from control group. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, n = 3.
Fig 7.
Inotodiol inhibits the growth of liver cancer tumors in mice.
(A-B) Photos of subcutaneous tumors in mice from each group. *significantly different from control group. *p<0.05, ***p<0.001, ****p<0.0001, n = 3.
Table 1.
The effect of inotodiol on the tumor volume and weight in tumor-bearing mice.
Table 2.
The effect of inotodiol on the body weight and organ mass of tumor-bearing mice.