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目的:通过分析microRNA-26a(miR-26a)mimics转染对人肝癌细胞株HepG2表达蛋白质组的影响,以确定miR-26a与肝癌发生发展的相关性。方法:常规培养人肝癌细胞株HepG2,经miR-26a mimics转染48 h后进行细胞周期分析,并裂解转染72 h的HepG2细胞提取蛋白,双向电泳分离,匹配对比各蛋白斑点的表达量,筛选主要差异表达蛋白进行质谱鉴定。结果:HepG2细胞经miR-26a mimics转染后细胞增殖受到抑制;其蛋白2-DE图谱与对照组比较,差异表达超过2倍的蛋白斑点有11个。其中,有3个蛋白斑点为表达上调,有8个蛋白斑点为表达下调。质谱鉴定为:膜联蛋白A1、过氧化物酶4、增殖细胞核抗原、载脂蛋白A1、细胞色素C氧化酶5a、细胞周期蛋白E2、磷酸核糖焦磷酸激酶3、周期素依赖性蛋白激酶1和磷脂酰乙醇胺结合蛋白。结论:miR-26a可能通过影响上述蛋白分子的表达,直接或间接地调控HepG2肝癌细胞的增殖、分化和死亡,以发挥其抗癌作用。
Objective: To analyze the effect of microRNA-26a (miR-26a) mimics transfection on proteome expression of human hepatoma cell line HepG2 to determine the correlation between miR-26a and the occurrence and development of hepatocellular carcinoma. METHODS: Human hepatocellular carcinoma cell line HepG2 was routinely cultured and transfected with miR-26a mimics for 48 h. Cell cycle analysis was performed. HepG2 cells transfected with 72 h of lysis were used for protein extraction. The proteins were separated by two-dimensional electrophoresis and matched to compare the expression of each protein spot. The major differentially expressed proteins were screened for mass spectrometry identification. RESULTS: The proliferation of HepG2 cells was inhibited after miR-26a mimics transfection. Compared with the control group, HepG2 cells showed 11 protein spots with more than 2-fold differential expression. Among them, there were 3 protein spots up-regulated and 8 protein spots down-regulated. Mass spectrometry identified as: annexin A1, peroxidase 4, proliferating cell nuclear antigen, apolipoprotein A1, cytochrome C oxidase 5a, cyclin E2, phosphoribosyl pyrophosphate kinase 3, cyclin-dependent protein kinase 1 And phosphatidylethanolamine binding protein. CONCLUSION: miR-26a may directly or indirectly regulate the proliferation, differentiation and death of HepG2 hepatocarcinoma cells by affecting the expression of the aforementioned protein molecules to exert its anticancer effect.