Apoptosis-inducing effect of grape seed proanthocyanidin extract on human ovary cancer SKOV3 cells and its mechanism / 重庆医科大学学报

ABSTRACT

Objective:To study the biological effects of grape seed extract(GSE) on the human ovary cancer cells and explore its molecular mechanism. Methods: Human ovary cancer cells line SKOV3 were cultured.They were put into 96-well plate and added with solutions of GSE of the terminal concentrations of 25,50,100,200,400 ?g/ml respectively,and 24,48,72 hours later cell growth curve was drawn. MTT assay was used for other SKOV3 cells co-incubated with GSE of the terminal concentration of 25,50,100,200,400 ?g/ml respectively so as to calculate the proliferation inhibition rate. SKOV3 cells were treated with GSE of the concentration of 25 ?g/ml for 24,48 hours respectively. Flowcytometry(FCM)was used to analyze the DNA cycle and TUNEL was used to calculate the apoptotic rate. Annexin-V labeling method was used to detect the positive rate of apoptotic cells. SKOV3 cells were treated with GSE of the concentration of 25 ?g/ml for 24,48,72 hours.Western blotting was used to detect the protein expression of caspase-3. Results:GSE dose-dependently inhibited the proliferation of the SKOV3 cells in a dose-dependent manner.Treated by GSE,the progress of cells at stage into G2/M stage was inhibited.Tunel showed that treated by GSE (25 ?g/ml)for 24、48 h the apoptotic rates of the cells were 31.98%and 45.78%, respectively. Annexin-V showed that after incubation with GSE (25 ?g/ml)for 12,24,48 h,the apoptotic rate were 6.71%,19.05%, 36.55%,respectively. Western blotting showed that the caspase-3 protein expression were up-regulated.ConlusionGSE inhibits the proliferation of malignant human ovary cancer cells and induces their apoptosis. Expression of caspase-3 protein may be related to cell growth inhibition and the apoptosis mediated by GSE in vitro.