Apoptosis of Human Ovarian Cancer Cells Induced by Paris Chinensis Dioscin via a Ca2+-Mediated Mitochondrion Pathway

Abstract

<b>Background:</b> Study of the mechanisms of apoptosis in tumor cells is an important field of tumor therapy andcancer molecular biology. Apoptosis triggered by activation of the mitochondrial-dependent caspase pathwayrepresents the main programmed cell death mechanism. The mitochondrial-dependent apoptosis pathway isactivated by various intracellular stresses that induce permeabilization of the mitochondrial membrane, leadingto cytochrome C release. This study was to investigate the anti-tumor effects of Dioscin from traditional Chineseanti-snake venom medicine Paris chinensis (PCD) and correlated mechanisms regarding apoptosis in humanovarian cancer SKOV3 cells. <br/><b>Methods</b>: Cell viability was analyzed by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay. Cell apoptosis was evaluated by flow cytometry and Laser ScanningConfocal Microscope (LSCM) using Annexin-V/PI staining. Intracellular calcium ions were detected usingfluorescence microscopy. The expression of apoptosis-related proteins cytochrome C and caspase-3 was measuredby immunohistochemical staining. <br/><b>Results</b>: PCD had an anti-proliferation effect on human ovarian cancerSKOV3 cells in a dose- and time-dependent manner. After treatment with PCD, the apoptotic rate significantlyincreased, and accompanied with the increased levels of caspase-3 and cytochrome C protein in SKOV3 cells.Morphological changes typical of apoptosis were also observed with LSCM by Annexin V/PI staining. Moreover,intracellular calcium accumulation occurred in PCD-treated cells. <br/><b>Conclusions</b>: The molecular determinants ofinhibition of cell proliferation as well as apoptosis of PCD may be associated with the activation of Ca2+-relatedm itochondrion pathway in SKOV3 cells.