Sinensetin flavone exhibits potent anticancer activity against drug-resistant human gallbladder adenocarcinoma cells by targeting PTEN/PI3K/AKT signalling pathway, induces cellular apoptosis and inhibits cell migration and invasion.

PURPOSE: The main focus of the current research work was to unveil the anticancer activity of the naturally occurring Sinensetin flavone against aggressive gall bladder cancer adenocarcinoma (GBAC) TJ-GBC2 cell line. Its effect of inducing apoptosis mediated via targeting PTEN/PI3K/AKT signalling pathway were also examined along with cell migration and invasion.

METHODS

Cell proliferation was tested by MTT cell viability assay. Fluorescence microscopy was utilized to carry out apoptosis related studies via DAPI staining along with flow cytometry using annexin V/propidium iodide (PI) assay. Further, western blotting analysis was carried out to examine the effects of Sinensetin on the expressions of apoptosis-related proteins and Bax Bcl-2 along with PTEN/PI3K/AKT signalling pathway. The impact of the test molecule on cell migration and invasion was studied through wound healing assay and transwell cell invasion assay respectively.

RESULTS

The results showed that Sinensetin treatment caused a significant retardation in cell viability, in a dose-dependent fashion. DAPI staining assay and annexin V/PI assay revealed that the cell viability of GBC cells was retarded due to induction of apoptosis. It was also associated with downregulation of Bcl-2 and upregulation of Bax levels. Further, wound healing assay and transwell cell invasion assay revealed that cell migration as well as cell invasion of cancer gallbladder cells was decreased in a concentration-dependent fashion. It was further seen that Sinensetin treatment resulted in inhibition of matrix metalloproteinase (MMP)-2 and enhancement of MMP-9 protein expressions. Results also showed that the tested molecule had the potential to inhibit PTEN/PI3K/AKT signalling pathway.

CONCLUSION

In conclusion, the current study indicated that Sinensetin flavone has the potential to be developed as a candidate drug against gallbladder adenocarcinoma provided more toxicological and in vivo studies are carried out.