目的：探讨miR-29b（microRNA-29b）的作用靶点及其对肝肿瘤细胞系Huh7细胞增殖及凋亡的干预作用。方法荧光定量PCR检测正常肝细胞系LO2及肝癌细胞系Huh7的miR-29b和Mcl-1表达水平。通过生物信息学分析预测Mcl-1基因是否受microRNA调控。将miR-29b模拟物通过Lipofectamine 2000顺时转染Huh7细胞，检测miR-29b对Mcl-1表达水平的影响。MTT法检测miR-29b模拟物转染对Huh7细胞增殖的影响。Annexin V/PI染色法检测miR-29b对Huh7细胞凋亡的影响。结果肝肿瘤细胞相比于正常肝细胞高表达Mcl-1（3.42依0.15比1.00依0.04，P<0.05）低表达miR-29b（0.43依0.04比1.00依0.06，P<0.05），在肝癌细胞中转染miR-29b可使Mcl-1的表达水平下降（0.37依0.03比1.03依0.07，P<0.05），转染miR-29b可抑制Huh7细胞的增殖（0.85依0.11比1.68依0.17，P<0.05）并诱导其发生凋亡[（28.4依2.10）%比（3.6依0.06）%，P<0.05]。结论 MiR-29b下调Huh7细胞Mcl-1蛋白的表达并诱导其发生凋亡。%Objective To investigate the therapeutic targets of miR-29b and its effect on proliferation and apoptosis of hepatocarcinoma cell line Huh7. Methods The expression of Mcl-1 and miR-29b in normal liver cell line LO2 and hepatocellular carcinoma cell line Huh7 was detected by qPCR. The putative binding sites of Mcl-1 gene was analyzed by using bioinformatic method. MiR-29b was transfected into Huh7 cells by Lipofectamine 2000, then the expression of Mcl-1 was detected. The cell growth was assessed by MTT after Huh7 cells transfected with miR-29b mimics. The apoptosis of cells was measured by Annexin V/PI staining. Results Compared with normal liver cells, Huh7 cells had higher expression of Mcl-1 (3.42±0.15 vs 1.00±0.04, P<0.05) and lower expression of miR-29b(0.43±0.04 vs 1.00±0.06, P<0.05). Transfection of miR-29b mimics suppressed Mcl-1 expression in Huh7 cells (0.37±0.03 vs 1.03±0.07, P<0.05) and inhibited the proliferation of Huh7 cells (0.85±0.11 vs 1.68±0.17, P<0.05) while inducing apoptosis in Huh7 cells [(28.4±2.1)% vs (3.6±0.06)%, P<0.05]. Conclusion MiR-29b down-regulates the expression of Mcl-1 protein inducing apoaptosis in Huh7 cells.
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