Apoptosis-Inducing Effect Of Grape Seed Proanthocyanidin Extract On Human Ovary Cancer Cell Line SKOV3 Cells And The Expression Of Apoptosis Associated Genes

Objective (1) This study was conducted to investigate the inhibitory effect of grape seed proanthocyanidin extract(GSPE) on the SKOV3 Cells proliferation by the vitro cell culture.at the same time,observe the diversify of morphologic.(2) Detect the apoptosis of SKOV3 cells induced by the different concentration GSPE.(3) through the extract from SKOV3 cells,to approach apoptosis-inducing effect of grape seed proanthocyanidin extract(GSPE)and the expression of apoptosis associated genes Survivin.(4) Analyze the Apoptosis-inducing Effect of GSPE on Human ovary cancer cell line SKOV3 Cells and the Expression of Apoptosis Associated Genes.Methods (1) in the study of vitro cell culture,The inhibition and cellular morphology by different concentration of GSPE in SKOV3 cells proliferation was analyzed by light microscopy,HE staining,MTT.(2) In the 2nd part's experiment,select the concentration of GSPE at 25μg/ml, Observe the rate of apoptosis by different concentration of GSPE through PI cell life cycle,Annexin V-FITC staining and TUNEL et al.(3) In the 3rd part's experiment,SKOV3 cells were treated with GSPE of the concentration of 25μg/ml for 24,48,72 hours,semi-quantitative RT-PCR was used to detect the mRNA expression of Surviving,and the protein expression of Survivin was detected by Westem blotting and immunohistochemistry.Result (1) Observe the SKOV3 cells incubated for 24,48,72 hours in presence of GSPE by light microscopy,HE staining,MTT.we acquire the following,The proliferation inhibition was shown as the lessened cell growth,decrease counting of mitotic cells.(2) A concentration-dependent and time-dependent proliferation apoptosis and survival were demonstrated in the SKOV3 cells incubated for 24,48,72 hours in presence of GSPE.and the protein expression of Caspase-3 was activated.(3) the semi-quantitation expression of Survivin proteins was decreased significantly by immunhistochemistry,Semiquantitative RT-PCR and Western blotting showed that the Survivin mRNA expression and protein expression were both down-regulated.Conclusion (1) GSPE inhibits the proliferation of malignant human ovary cancer cells in the study of vitro cell culture.(2) GSPE and induces the malignant human ovary cancer SKOV3 cells apoptosis.(3) Expression of Survivin mRNA and protein may be related to cell growth inhibition and to the apoptosis mediated by GSPE in vitro.(4) GSPE can induce SKOV3 cells apoptosis.The molecular mechanism might be related to up-regulating expression of Caspase-3 and down-regulating expression of Survivin genes.