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目的研究减数分裂重组蛋白11(MRE11)基因沉默对5-氟尿嘧啶(5-FU)耐药的肝癌细胞株Bel7402/5-FU的化疗敏感性及耐药相关蛋白1(MRP1)表达的影响,探讨MRE11与肝癌耐药性的关系。方法采用阳离子脂质体法将shMRE11干扰质粒转染BEL7402/5-FU细胞,实时定量PCR(qRT-PCR)及Western blot法检测沉默效率;MTT法检测转染后BEL7402/5-FU细胞对化疗药物顺铂(DDP)、丝裂霉素(MMC)、阿霉素(ADM)、5-FU的敏感性,qRT-PCR及Western blot法分别检测转染后BEL7402/5-FU细胞中耐药相关蛋白MRP1 mRNA水平及蛋白水平的变化。结果 qRT-PCR及Western blot法检测显示MRE11mRNA及蛋白水平的沉默效率分别为78.0%、56.1%。MTT结果显示,shMRE11转染BEL7402/5-FU细胞后,shMRE11实验组对DDP、MMC、ADM、5-FU的IC50均低于对照组(P<0.05)。qRT-PCR及Western blot法检测结果显示shMRE11实验组MRP1mRNA及蛋白水平的表达与对照组相比均有所降低(P<0.05)。结论 shMRE11能够提高肝癌耐药细胞BEL7402/5-FU对化疗药物的敏感性,降低耐药相关蛋白MRP1的表达。
Objective To investigate the effect of MRE11 gene silencing on the chemosensitivity and drug resistance-related protein 1 (MRP1) expression of 5-fluorouracil (5-FU) resistant hepatocellular carcinoma cell line Bel7402 / 5- To investigate the relationship between MRE11 and drug resistance of liver cancer. Methods The silencing efficiency of shMRE11 plasmid was transfected into BEL7402 / 5-FU cells by cationic liposome method. The efficiency of silencing was evaluated by real-time quantitative PCR (qRT-PCR) and Western blot. The effect of BEL7402 / 5- The drug resistance of BEL7402 / 5-FU cells was detected by qRT-PCR and Western blot after the drug sensitivity of DDP, ADM, ADM and 5-FU Related protein MRP1 mRNA levels and protein levels. Results The results of qRT-PCR and Western blot showed that the silencing efficiencies of MRE11 mRNA and protein were 78.0% and 56.1%, respectively. The results of MTT assay showed that the IC50 of shMRE11 transfected BEL7402 / 5-FU cells were lower than that of the control group (P <0.05). The results of qRT-PCR and Western blot showed that the expression of MRP1 mRNA and protein in shMRE11 group was lower than that in control group (P <0.05). Conclusion shMRE11 can improve the chemosensitivity of BEL7402 / 5-FU cells, and decrease the expression of MRP1.