摘要目的 探讨三羟异黄酮对胰腺癌BxPC3细胞Notch-1和Hedgehog信号通路分子SHH和HHIP表达以及对细胞周期和增殖的影响.方法 体外培养胰腺癌BxPC3细胞,三羟异黄酮作用BxPC3细胞后,提取总RNA和蛋白质.RT-PCR方法检测Notch-1 mNRA、SHH mRNA和HHIP mRNA表达,Western blotting方法检测Notch-1和SHH蛋白的表达.MTT方法检测BxPC3细胞的增值,流式细胞术和PI单染法分析细胞的周期.结果 20μg/ml的三羟异黄酮处理BxPC3 48 h后,细胞增殖抑制率为(67.17±2.32)%.Notch-1 mRNA表达从2.454±0.068下调到1.304±0.169;SHH mRNA表达从0.959±0.023下调到0.472±0.077;而HHIP mRNA表达从0.625±0.158增加到1.761±0.121.Notch-1蛋白表达从1.361±0.109下调到0.760±0.114;SHH蛋白表达从0.265±0.018下调到0.129±O.013.(52.77±9.47)%的细胞被阻滞在G2/M期.结论 三羟异黄酮能同时抑制Notch-1的表达和阻断Hedgehog信号通路,并抑制胰腺癌细胞的增殖.

abstractsObjective To investigate the effect of genistein on Notch-1, SHH and HHIP gene expression and on the cell cycle and proliferation of of BxPC3 cells. Methods Human pancreatic cancer cell line BxPC3 was cultured. The BxPC3 cells were treated with genistein and then the total RNA and protein were extracted. RT-PCR was used to detect the expression of Notch-1 mRNA, SHH mRNA and HHIP mRNA. Noteh-1 and SHH protein was determined by western blotting. MTT assay was used to detect proliferation of BxPC3 cells. The cell cycle of BxPC3 cells was measured by Propidium iodide (PI) and flow cytometry. Results The inhibiting rate was 67.17%±2.32% when BxPC3 cell lines were treated by 20μg/ml genistein for 48 hours. Notch-1 mRNA was down-regulated from 2.454±0.068 to 1.304±O.169 ; SHH mRNA was down-regulated from 0.959±0.023 to O.472±0.077 ; HHIP mRNA was up-regulated from 0.625±O.158 to 1.761±0.121. Notch-1 protein expression was down-regulated from 1.361±0.109 to 0.760±0.114; SHH protein expression was down-regulated from 0.265±0.018 to 0.129±0.013. (52.77±9.47)% cells were hindered in G2/M stage. Conclusions Genistein could down-regulate Notch-1 expression and inactivate Hedgehog signaling pathway and inhibit the proliferation of pancreatic cancer cells.