摘要
目的观察三结构域蛋白59(TRIM59)在人肝细胞癌(HCC)细胞中的表达变化,及其对细胞增殖和p53蛋白表达的影响。方法选取6种HCC细胞系(Huh7、Hep3B、HepG2、SK-Hep1、BEL7402、SMMC7721)及人正常肝细胞系LO2为研究对象,采用qRT-PCR和Western blotting法分别检测细胞中的TRIM59 mRNA和蛋白,选择预实验中TRIM59 mRNA和蛋白表达水平相对较高的HCC细胞进行下一步实验。将HepG2细胞分为shTRIM59组和shControl组,shTRIM59组感染TRIM59慢病毒,shControl组转染TRIM59慢病毒阴性对照。将Huh7细胞分为pcDNA3.1-TRIM59组和pcDNA3.1-Vector组,分别转染pcDNA3.1-TRIM59质粒和pcDNA3.1-Vector对照质粒。采用集落形成实验检测HepG2、Huh7的细胞增殖能力,采用Western blotting法检测细胞中的p53蛋白。结果各HCC细胞中TRIM59 mRNA及蛋白相对表达量均高于LO2细胞,HepG2及Huh7细胞中TRIM59的表达高于其他HCC细胞系(P均<0.05)。shTRIM59组HepG2细胞集落形成数低于shControl组,p53蛋白相对表达量高于shControl组(P均<0.05)。pcDNA3.1-TRIM59组Huh7细胞集落形成数高于pcDNA3.1-Vector组,p53蛋白相对表达量低于pcDNA3.1-Vector组(P均<0.05)。结论HCC细胞中TRIM59 mRNA及蛋白高表达;上调TRIM59表达,HCC增殖能力增强、p53蛋白低表达,下调TRIM59表达则HCC增殖能力减弱、p53蛋白高表达;TRIM59对HCC细胞增殖的影响可能是通过调节p53蛋白表达实现的。
Objective To observe the expression of tripartite-motif 59(TRIM59)in the human hepatocellular carcinoma(HCC)cells and its effect on cell proliferation and p53 expression.Methods Six kinds of HCC cell lines(Huh7,Hep3B,HepG2,SK-Hep1,BEL7402,SMMC7721)and human normal liver cell line LO2 were selected as subjects.The expression of TRIM59 mRNA and protein in experimental cells were detected by Western blotting and qRT-PCR.HCC cells with relatively high levels of TRIM59 mRNA and protein expression were selected for the next experiment.HepG2 cells were divided into two groups:the shTRIM59 group which was transfected with TRIM59 lentivirus and the shControl group which was transfected with lentivirus negative-shRNA.Huh7 cells were divided into the pcDNA3.1-TRIM59 group and pcDNA3.1-Vector group,and were transfected with pcDNA3.1-TRIM59 plasmid and pcDNA3.1-Vector control plasmid,respectively.Cell proliferative capacity of HepG2 and Huh7 cells were detected by colony formation experiment and the expression of p53 protein was detected by Western blotting.Results The relative expression levels of TRIM59 mRNA and protein were higher in the HCC cell lines than in the human normal liver cell line LO2.The expression of TRIM59 in HepG2 and Huh7 cells was higher than that of the other HCC cell lines(P<0.05).The shTRIM59 group had a smaller number of colony formation than the shControl group in HepG2 cells,and had a higher relative expression of p53 protein than the shControl group(both P<0.05).The pcDNA3.1-TRIM59 group had a higher number of colony formation than the pcDNA3.1-Vector group in Huh7 cells,and had a lower relative expression of p53 protein than the pcDNA3.1-Vector group(both P<0.05).Conclusions TRIM59 mRNA and protein are highly expressed in the HCC cells.When TRIM59 is up-regulated in HCC cells,the cell proliferative capacity is enhanced,and the expression of p53 decreases;when TRIM59 is down-regulated,the cell proliferative capacity is weakened,and the expression of p53 increases.The effect of TRIM59 on HCC cell proliferation might be achieved by regulating p53 expression.
作者
卢光辉
孙纲
刘子源
徐鹏
邵润东
陈琦
鲍丰羽
LU Guanghui;SUN Gang;LIU Ziyuan;XU Peng;SHAO Rundong;CHEN Qi;BAO Fengyu(The No.967 Hospital of PLA Joint Logistics Support Force,Postgraduate Training Base of Jinzhou Medical University,Dalian 116021,China;不详)
出处
《山东医药》
CAS
2020年第8期44-47,共4页
Shandong Medical Journal
