Inhibitive Effect And Mechanism Of Grape Seed Procyanidin Extract On Human Bladder Cancer Cells

Partâ… Effect of growth inhibition of grape seed procyanidinextract on human bladder cancer BIU87 cellsObjective To investigate the effect of growth inhibition of grape seed procyanidinextract(GSPE) on human bladder cancer BIU87 cells and human primary skin fibroblasts.Methods BIU87 cells and human primary skin fibroblasts were cultured with differentconcentrations of GSPE(12.5, 25, 50, 100 and 200μg/ml) for 24 h. The morphologicalchanges of BIU87 cells and human primary skin fibroblasts were observed, the cellviabilities were determined by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazoliumbromide(MTT) assay.Results Compared with the control group, 50, 100 and 200μg/ml of GSPE causedcharacteristic morphological changes in BIU87 cells. The human primary skin fibroblastsshowed no morphological changes in all groups. The viability of the BIU87 cells incubatedwith GSPE at concentrations of 12.5, 25, 50, 100 and 200μg/ml for 24 h was 99.55±0.50%(n=18, P＞0.05), 99.16±0.63% (n=18, P＞0.05), 86.98±1.48%(n= 18, P＜0.01), 68.18±2.06%(n=18, P＜0.01) and 51.42±1.78% (n=18, P＜0.01) of the control value, respectively. 50, 100and 200μg/ml of GSPE inhibited the growth of BIU87 cells, while no effect on humanprimary skin fibroblasts.Conclusions GSPE, in some concentration extent, inhibites the growth of human bladdercancer BIU87 cells, and has no influence on the growth of human primary skin fibroblasts. Partâ…¡Inhibition of cell cycle by grape seed procyanidin extractin human bladder cancer BIU87 cellsObjects To study the effect of grape seed procyanidin extract (GSPE) on cell cycle inhuman bladder cancer BIU87 cells and investigate its molecular mechanism.Methods BIU87 cells were treated with different concentrations(50, 100 and 200μg/ml) ofGSPE and cultured for 24 h in vitro while untreated group as control, flow cytometry wasused to evaluate cell cycle, and RT-PCR and Western blot were used to detect the mRNAand protein expression of cyclin D1 and CDK4.Results We found that GSPE inhibited the cell growth through cell cycle arrest at G₁ phraseby a dose-dependent manner. Semiquantitated RT-PCR and Western blot analyses indicatedthat GSPE decreased cyclin D1 and CDK4 expression significantly (P＜0.01).Conclusions GSPE induces cell cycle arrest in BIU87 cells in vitro, and the effect mayberelated with its down-regulation of cyclin D1 and CDK4. Partâ…¢Grape seed procyanidin extract induces apoptosis inhuman bladder cancer BIU87 cellsObjects To study the effect of grape seed procyanidin extract (GSPE) on cell apoptosis inhuman bladder cancer BIU87 cells and investigate its molecular mechanism.Methods BIU87 cells were treated with different concentrations(50, 100 and 200μg/ml) ofGSPE and cultured for 24 h in vitro while untreated group as control, MTT[3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide] assay, hoechst 33258 stainning,flow cytometry, RT-PCR and Western blot were used to detect the apoptotic induction effectof GSPE on BIU87 cells.Results We found that GSPE induced cell apoptosis in BIU87 cells by a dose-dependentmanner. Semiquantitated RT-PCR and Western blot analyses indicated that GSPE increasedthe expression of caspase-3 and decreased the expression of survivin (P＜0.01).Conclusions GSPE induces apoptosis in BIU87 cells in vitro, and the effect maybe relatedwith its down-regulation of survivin. Partâ…£Inhibitive effect of combining mitomycin C with grapeseed procyanidin extract on human bladder cancer BIU87 cellsObjective To explore the anti-cancer effects of mitomycin C (MMC) and grape seedprocyanidin extract (GSPE) on human bladder cancer BIU87 cells.Methods The 3-[4,5-dimethylthiazol-2-y1]-2,5-diphenyltetrazolium(MTT) method wasused to detect the inhibition rates of MMC and GSPE single or combined, the combinationindex(CI) method of Chou-Talalay was used to analyze the effect of combined applicationquantitatively.Results The anti-cancer effects of MMC and GSPE were enhanced with the increasing ofdrug concentration when used alone. The media-effect concentrations were 15.422 and195.865μg/ml respectively. MMC was synergetic with GSPE (CI＜1) at low concentrations(0＜fa＜37%), while antagonist (CI＞1) at high concentrations (37%＜fa＜100%) when usedcombined. The media-effect concentration was 109.496μg/ml (MMC 8.111μg/ml andGSPE 101.385μg/ml).Conclusions The effect of combination between MMC and GSPE is synergetic at lowconcentrations, while antagonist at high concentrations.