间皮素基因沉默对胰腺癌化疗药物作用的影响

摘要

Objective To investigate the effect of mesothelin silencing on the inhibitory effect of chemotherapeutic drugs in pancreatic cancer.Methods PANC-1 cells were treated with transcient transfection of siRNA of mesothelin.Flow cytometry and fluorescent microscopy were used to detect the efficiency of the transfection.The expression of mesothelin was evaluated hy Western hlot analysis.Then the cells were treated with mesothelin siRNA with or in the absence of different concentration of chemotherapeutic agents gemcitabine, cisplatin, cucurbitacin E or doxorubicin.After 48 hours, the cell proliferation was tested by MTT assay.Results Flow cytometry and fluorescence microscopy showed that a fluorescent labeled cells accounted for the most of the total and proved the successful transfection.Western blot analysis showed that silencing of mesothelin gene suppressed the expression of the mesothelin.The proliferation of PANC-1 was significantly inhibited after the mesothelin gene silencing (P ＜ 0.05 ) .After mesothelin silencing, the growth inhibition rate of the drugs increased significantly at the low concentration.Conclusions Gene silencing of mesothelin inhibits the growth of pancreatic cancer PANC-1 in vitro.Silencing of mesothelin in combination with chemotherapeutic drugs can result in synegistic inhibitory effect of pancreatic cancer cells.%目的 研究胰腺癌特异性抗原间皮素基因沉默对胰腺癌化疗药物的影响.方法 以间皮素siRNA和荧光标记的siRNA分别通过脂质体瞬时转染入胰腺癌PANC-1细胞,流式细胞仪和倒置相差荧光显微镜检测转染效率,Western blot检测间皮索基因沉默的效果.以胰腺癌化疗药物吉西他滨、顺铂、阿霉素和葫芦素E分别处理间皮素基因沉默后的PANC-1细胞,继续培养48 h后,MTT法检测细胞增殖情况.结果 流式细胞仪和倒置荧光显微镜检测结果证明细胞转染成功;Westem blot检测结果显示间皮素基因沉默后,间皮素蛋白基本无表达;MTT结果显示间皮素基因沉默后细胞增殖减少(P<0.05);间皮素基因沉默后,能增强化疗药物的作用.结论 间皮素基因沉默对胰腺癌化疗药物的作用具有增强作用.