Effect of Scopoletin on Apoptosis and Cell Cycle Arrest in Human Prostate Cancer Cells In vitro
Purpose: To investigate the anticancer activity of scopoletin against human prostate cancer.
Methods: The anticancer activity of scopoletin was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5- diphenyltetrazolium bromide (MMT) assay. Flow cytometry using propidium iodide and annexin V-FITC was employed to study apoptosis and cell cycle analysis. Hoechst 33258 staining was used to assess the effect of scopoletin on cell morphology and apoptotic body formation in human prostate carcinoma (LNCaP) cells via Florescence microscopy and finally Western blotting was used to evaluate the effect of scopoletin on cyclin D1 and cyclin B1 expressions.
Results: Scopoletin induced a dose-dependent growth inhibition in LNCaP prostate cancer cells. It induced G2/M phase growth arrest and led to an increase in the sub-G0/G1 cell population after treatment with increasing doses compared to control cells, scopoletin treatment resulted in cell shrinkage along with membrane blebbing which are characteristic features of cell apoptosis. Approximately 15.45, 32.6 and 21.71 % of the cells underwent early apoptosis after treatment with 40, 80 and 100 μM of scopoletin respectively. Cyclin D expression diminished in a concentration-dependent manner when LNCaP cells were treated with different concentrations of scopoletin.
Conclusion: These results reveal that scopoletin may be used as a natural chemotherapeutic agent against prostate cancer.
Keywords: Prostate cancer, Apoptosis, Cell cycle analysis, Scopoletin, Flow cytometry, Fluorescence microscopy