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目的探讨多药耐药(MDR1)基因沉默对大肠癌耐药细胞系LoVo/5-Fu耐药性的逆转作用。方法构建靶向MDR1的短发夹RNA(EASY-shRNA-MDR1)干扰质粒转染LoVo/5-Fu,实时荧光定量PCR(Realtime-PCR)在mRNA水平检测抑制效果,噻唑蓝(MTT)法检测细胞对5-Fu的敏感性,流式细胞仪检测细胞凋亡情况。结果与未转染组相比,EASY-shRNA-MDR1组细胞MDR1 mRNA表达明显下调(P<0.05);细胞对5-Fu的IC50及RI显著降低(P<0.05),敏感性的相对逆转率为74.7%;凋亡率明显升高(P<0.05)。结论 EASY-shRNA-MDR1可有效抑制大肠癌耐药细胞中MDR1的表达,进而降低了大肠癌耐药株对5-Fu的耐药性。
Objective To investigate the reversal effect of multidrug resistance (MDR1) gene silencing on the drug resistance of multidrug-resistant human LoVo / 5-Fu cells. Methods Real-time quantitative PCR (Realtime-PCR) was used to detect the inhibitory effect of LoVo / 5-Fu on the mRNA level of MDR1 and EASY-shRNA-MDR1. MTT assay Sensitivity of cells to 5-Fu, flow cytometry to detect apoptosis. Results Compared with untransfected cells, the expression of MDR1 mRNA in EASY-shRNA-MDR1 group was significantly down-regulated (P <0.05); the IC50 and RI of 5-Fu cells were significantly decreased (P <0.05) Was 74.7%; the apoptosis rate was significantly increased (P <0.05). Conclusion EASY-shRNA-MDR1 can effectively inhibit the expression of MDR1 in multidrug-resistant cells of colorectal cancer and reduce the drug resistance of 5-Fu in multidrug-resistant colon cancer cells.