Effect Of Compound K On Apoptosis Of Human Ovarian Cancer CAOV3Cells

Objective:In this study we want to examine the effect of CK on ovarian cancer, and explore the underlying mechanism. The results obtained this study will provided theoretical evidence for developing an anti-cancer drug in the future.Method:1、MTT assay was used to detect the effect of CK on proliferation of human ovarian carcinoma cells, and to measure half inhibitory concentration (IC50);2、DAPI nuclear staining was used to detect apoptotic morphology of cells;3、Western blot was applied to show the status of Poly ADP-Ribose Polymerase (PARP), which is specificity cleaved after the Caspase-3activation;4、Cell free Caspase activity assay was applied to determine Caspase-3,-8,-9activity in apoptotic cells.Results:1、CK effectively inhibited CAOV3cell proliferation in a dose dependent manner, with IC50value of3.6μg/ml;2、CK induced apoptosis in CAOV3cells, evidenced by morphological characteristics of apoptosis, such as cell rounding, membrane bleb bing, and nuclei condensation and fragmentation;3、Western blot analysis demonstrated that PARP, a specific substrate of Caspase-3was cleaved after treating CAOV3cells with2.5, and5g/ml CK, and this cleavage was increased in a dose-dependent manner indicating Caspase-3activation was involved in CK induces Caspase-3involved apoptosis.4、Cell free Caspase activity assay showed that Caspase-3,-8,-9were activated in CK treated CAOV3cells. These data suggest that CK induces CAOV3cell apoptosis via both intrinsic and extrinsic apoptosis pathway.Conclusion:1、CK have toxicity impact on Ovarian cancer CAOV3cells.2、This effect is achieved by apoptosis.3、The process of apoptosis is regulated by a variety of cell signaling,according to the signal source to the apoptosis pathway is divided into two categories:one is extrinsic pathway triggered by interactions between cytokines and death receptors, resulting in activation of Caspase-8;the other is intrinsic pathway can be initiated by mitochondrial membrane demilitarization that facilitates the release of cytochrome c,wich is then associated with Apaf-1and Caspase-9to promote Caspase activation.