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.Conlusion:GSE 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.
ABSTRACT
Objective To study the biological effects of grape seed proanthocyanidin extract (GSPE) on human ovary cancer cells and to explore the molecular mechanism. Methods Human ovary cancer cell line SKOV3 were co-cultured with GSPE solution of the terminal concentrations of 25,50,100,200,400 ?g/ml respectively in 96-well plate. At 24,48,72 h after coculture,the following parameters were detected: cell growth curve,the inhibition rate of SKOV3 cells by MTT assay,DNA cycle by FCM,the apoptosis of SKOV3 by TUNEL and Annexin-V labeling method,the mRNA and protein expressions of survivin by semi-quantitative RT-PCR and Western blotting,respectively. Results GSPE dose-dependently inhibited the proliferation of the SKOV3 cells. Treated by GSPE,the progress of SKOV3 cells at S stage into G/M stage was inhibited. TUNEL showed that treated by 25 ?g/ml GSPE for 24,48 h,the apoptotic rates of SKOV3 cells were 31.98%,45.78% respectively. Annexin-V showed that after incubation with 25 ?g/ml GSPE for 24 h,the apoptotic rate was 14.68%. The survivin mRNA and protein expressions were both down-regulated. Conclusion GSPE inhibits the proliferation of malignant human ovary cancer cells and induces their apoptosis. Expression of survivin mRNA and protein may be related to cell growth inhibition and to the apoptosis mediated by GSPE in vitro.