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目的:探讨MiRNA-122对肝癌耐药细胞株BEL-7402/氟尿嘧啶(fluorouracil,FU)的FU敏感性的影响及其作用机制。方法:构建miR-122及其阴性对照空载体并稳定转染至BEL-7402/FU细胞中,Western免疫印迹检测miR-122转染组(n=3)、阴性空载体转染组(n=3)和未转染组(n=3)中与耐药相关的Bcl-2和Bcl-XL蛋白表达水平。用3-(4,5-二甲基-2-噻唑)-2,5-二苯基溴化四氮唑噻唑[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide,MTT)]法检测三组细胞对FU敏感性。结果:与未转染组、阴性空载体转染组比较,miR-122转染组Bcl-2和Bcl-XL蛋白表达明显降低(P<0.05)。MTT结果显示:在不同浓度FU处理下,miR-122转染组的细胞抑制率较未转染组和阴性空载体转染组高(P<0.05)。结论:miR-122能特异性地下调Bcl-2和Bcl-XL表达,可增加BEL-7402/FU细胞对FU敏感性。
Objective: To investigate the effect of MiRNA-122 on FU sensitivity of drug-resistant hepatocarcinoma cell line BEL-7402 / fluorouracil (FU) and its mechanism. METHODS: miR-122 and its negative control empty vector were constructed and stably transfected into BEL-7402 / FU cells. Western blotting was used to detect miR-122 transfected group (n = 3) 3) and non-transfected group (n = 3), the expression of Bcl-2 and Bcl-XL protein in drug-resistant group. (4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazoliumbromide using 3- (4,5-dimethyl- , MTT)] method to detect the sensitivity of three groups of cells to FU. Results: The expressions of Bcl-2 and Bcl-XL in miR-122 transfected group were significantly lower than those in untransfected group and negative transfected group (P <0.05). The results of MTT assay showed that the cell inhibition rate of miR-122 transfected group was higher than that of untransfected group and negative empty vector transfected group (P <0.05). Conclusion: miR-122 can specifically down-regulate the expressions of Bcl-2 and Bcl-XL, and increase the sensitivity of BEL-7402 / FU cells to FU.