慢病毒介导RNAi抑制胃癌细胞CDH17基因的表达对化疗药物敏感性的影响

Lentivirus-mediated RNA Interference Suppresses CDH17 Expression and Enhances Chemosensitivity in Gastric Cancer Cells

[目的]探讨慢病毒介导的RNA干扰(RNAi)沉默肝肠—钙黏连蛋白(CDH17)基因的胃癌细胞SGC-7901对化疗药物敏感性的影响。[方法]将制备好的siRNA-CDH17慢病毒表达载体稳定转染SGC-7901细胞72h后,采用实时定量PCR法检测转染后SGC-7901细胞中CDH17基因的表达水平;Western blot检测CDH17蛋白的表达水平;利用流式细胞仪检测细胞周期,MTT法和平板克隆形成实验检测CDH17基因沉默后胃癌细胞SGC-7901对氟尿嘧啶(5-FU)的敏感性。[结果]Real-time PCR及Western blot检测结果表明CDH17 mRNA及蛋白在SGC-7901细胞中表达下调;实验组SGC-7901细胞周期明显阻滞在G0/G1期,S期相对减少;5-FU处理SGC-7901细胞48h后,MTT法和平板克隆形成实验显示实验组增殖抑制显著高于另两组,差异有统计学意义(P<0.001)。CDH17基因沉默的SGC-7901细胞对5-FU的敏感性显著增强。[结论]以CDH17基因为靶标的RNA干扰能下调SGC-7901细胞中CDH17的表达,从而增强其对5-FU的化疗敏感性。

[Purpose] To investigate the effects of lentivirus-mediated RNA interference（RNAi） targeting CDH17（liver-intestine cadherin） on the chemosensitivity to 5-FU of gastric cancer cell line SGC-7901.[Methods] SGC-7901 cells were steady transfected with lentiviruses delivering small interfering RNA（siRNA） against CDH17 gene.The expression levels of CDH17 mRNA and protein were detected by real-time PCR and Western blot,respectively.Flow cytometry was used to detect the cell cycle.Sensitivity to 5-FU after transfection were examined by MTT and colony formation assay.[Results] In SGC-7901 cells,the expression levels of CDH17 mRNA and protein decreased significantly after transfection.The cell number in G0/G1 phase increased while that in S phase decreased in experimental group cells.Forty-eight hours after treatment with 5-FU,the growth inhibition rate in experimental group was significantly higher than that in the other two groups（P0.001）.The sensitivity of SGC-7901 cells to 5-FU increased significantly after transfection.[Conclusion] Lentivirus-mediated RNAi targeting CDH17 can effectively suppress the expression of CDH17 in SGC-7901 cells and enhance the chemosensitivity of the cells to 5-FU.