lncRNA TUG1对肝癌细胞增殖及自噬水平的影响

The effect of lncRNA TUG1 on proliferation and autophagy in HCC cells

目的:检测长链非编码RNA牛磺酸上调基因(taurine upregulated gene 1,TUG1)在肝癌组织中的表达情况,证实TUG1对肝癌细胞增殖和自噬能力的影响。方法:实时定量PCR检测50例肝癌组织标本以及相应癌旁组织中TUG1的表达水平以及在肝癌细胞系中的表达情况。通过将构建的TUG1过表达质粒转染肝癌细胞系SMMC-7721和HepG2,验证转染效率并采用CCK-8法检测肝癌细胞增殖情况;Western blot检测自噬标志蛋白(LC3-Ⅱ/Ⅰ)表达变化;通过实时定量PCR筛选表达差异最显著的自噬相关基因并采用Western blot验证。结果:肝癌组织中TUG1的表达显著高于癌旁组织,差异有统计学意义(P <0. 05)。TUG1能够显著促进肝癌细胞增殖能力(P <0. 05);并且过表达TUG1后增加了LC3-I向LC3-Ⅱ的转化,促进自噬水平的发生(P <0. 05);TUG1能够显著上调ATG7的基因和蛋白的表达。结论:在肝癌组织中TUG1表达高于癌旁组织;在体外细胞实验中发现,TUG1能够促进肝癌细胞增殖能力,并通过上调ATG7的表达诱导自噬的发生。

Objective:To investigate the expression level of long non-coding RNA TUG1(taurine upregulated gene 1,TUG1) in HCC tissues and confirm the effect of TUG1 on proliferation and autophagy in HCC cells.Methods:Real-time PCR method was used to detect the expression of TUG1 in 50 HCC samples,paracancerous tissues and HCC cell lines.The transfection efficiency was detected by TUG1 overexpression plasmid transfecting the HCC cell lines SMMC-7721 and HepG2.Moreover,CCK-8 assay was conducted to detect the proliferation of hepatocellular carcinoma cells.The expression levels of autophagy marker protein(LC3-Ⅱ/I) were detected by Western blot.Besides,Western blot and Real-time PCR were performed to screen and verify the most significant autophagy-related gene.Results:The expression levels of TUG1 in HCC tissues were significantly higher than paracancerous tissues(P<0.05).TUG1 significantly promoted the proliferation of hepatoma cells(P<0.05).Overexpressed TUG1 increased the conversion of LC3-I to LC3-Ⅱ and promoted the level of autophagy(P<0.05).TUG1 significantly facilitated the expression of ATG7 gene and protein.Conclusion:TUG1 is upregulated in HCC tissues compared to paracancerous tissues.TUG1 can promote the proliferation of hepatoma cells and induce autophagy by up-regulating the expression of ATG7 in vitro.