Nael Abutaha1. 1. Bioproducts Research Chair, Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia.
Abstract
PURPOSE: Cancer is a major health problem worldwide. There is a continuous need to search for safer and more effective alternatives to overcome the side effects and resistance of the chemotherapeutic agents. Therefore, in this study we investigated the antiproliferative activity and the apoptotic potential of Withania somnifera (W. somnifera). METHODS: W. somnifera was extracted with methanol and then solvent partitioned by sequential extractions with hexane, dichloromethane and ethyl acetate. Each extract was assayed for antiproliferative activity against different cancer cell lines using MTT assay. The nuclear morphology of HepG2 cells was investigated by DNA-binding fluorescent dye (Hoechst 33342 stain). The percentage of viability, death and apoptosis were evaluated by the Tali(TM) Image-based cytometer using annex-in-V/PI (propidium iodide). A chromatographic fingerprint was constructed using high performance liquid chromatography (HPLC). RESULTS: The most potent anticancer activity of the crude extract was against HepG2 cell line (LC50=164.7μg/ml). Dichloromethane fraction showed remarkable changes in the chromatin structure i.e., fragmentation, uniform condensation. Of the HepG2 cells 43.6% were apoptotic when treated with dichloromethane fraction for 24 hrs at 95μg/ml concentration. HPLC showed the presence of a major peak at 11.85 min. CONCLUSION: W. somnifera may have the potential to serve as a template for future anticancer drug development. However, further investigation is required to identify the active compound/s.
PURPOSE:Cancer is a major health problem worldwide. There is a continuous need to search for safer and more effective alternatives to overcome the side effects and resistance of the chemotherapeutic agents. Therefore, in this study we investigated the antiproliferative activity and the apoptotic potential of Withania somnifera (W. somnifera). METHODS:W. somnifera was extracted with methanol and then solvent partitioned by sequential extractions with hexane, dichloromethane and ethyl acetate. Each extract was assayed for antiproliferative activity against different cancer cell lines using MTT assay. The nuclear morphology of HepG2 cells was investigated by DNA-binding fluorescent dye (Hoechst 33342 stain). The percentage of viability, death and apoptosis were evaluated by the Tali(TM) Image-based cytometer using annex-in-V/PI (propidium iodide). A chromatographic fingerprint was constructed using high performance liquid chromatography (HPLC). RESULTS: The most potent anticancer activity of the crude extract was against HepG2 cell line (LC50=164.7μg/ml). Dichloromethane fraction showed remarkable changes in the chromatin structure i.e., fragmentation, uniform condensation. Of the HepG2 cells 43.6% were apoptotic when treated with dichloromethane fraction for 24 hrs at 95μg/ml concentration. HPLC showed the presence of a major peak at 11.85 min. CONCLUSION:W. somnifera may have the potential to serve as a template for future anticancer drug development. However, further investigation is required to identify the active compound/s.