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[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。
[Objective] To investigate the effect of lentivirus-mediated RNA interference (RNAi) silencing SGC-7901 on the sensitivity of chemotherapeutic drugs to hepatecto-cadherin (CDH17) gene. [Method] The constructed siRNA-CDH17 lentiviral vector was stably transfected into SGC-7901 cells for 72 hours. The expression of CDH17 gene in SGC-7901 cells was detected by real-time quantitative PCR. Western blot was used to detect CDH17 protein The sensitivity of SGC-7901 cells to 5-fluorouracil (5-FU) was tested by MTT assay and plate clone formation assay by flow cytometry. [Results] The results of Real-time PCR and Western blot showed that the expression of CDH17 mRNA and protein were down-regulated in SGC-7901 cells. The cell cycle arrest of SGC-7901 cells in G0 / G1 phase and the S phase in 5-FU After treatment of SGC-7901 cells for 48h, MTT assay and plate clone formation assay showed that the inhibition of proliferation in SGC-7901 cells was significantly higher than that in the other two groups (P <0.001). The sensitivity of SGC-7901 cells to CDH17 gene silencing to 5-FU was significantly increased. [Conclusion] The RNA interference of CDH17 gene can down-regulate the expression of CDH17 in SGC-7901 cells and enhance the chemosensitivity to 5-FU.