不同转移潜能肝癌细胞株miRNAs表达谱的差异性研究

Research of differences of miRNAs expression profiles in different metastatic potential HCC cell lines

目的筛选不同转移潜能肝癌细胞株的microRNA(miRNA)差异表达谱,并预测分析差异表达miRNAs调控的靶基因与功能。方法提取总RNA进行miRNA芯片分析,通过分析MHCC-97H(高转移)、Hep3B(不转移)两种不同转移潜能肝癌细胞株与正常肝细胞L02比较,以及两种肝癌细胞株相互比较的miRNA差异表达谱,筛选出差异表达明显的miRNAs进行qRT-PCR验证,利用4个靶基因预测软件(Target Scan、miRanda、miRWalk、miRDB)进行靶基因预测,并通过生物信息学分析来探讨候选靶基因的生物学功能。结果与正常肝细胞L02相比,肝癌细胞株MHCC-97H、Hep3B中的miR-192-5p、miR-215-5p表达均明显上调,而miR-130a-3p、miR-196a-5p则明显降低;与不转移肝癌细胞株Hep3B相比,miR-224-5p在肝癌细胞株MHCC-97H中明显上调,而miR-146a-5p、miR-483-3p、miR-200b-3p则明显降低。qRT-PCR验证结果与芯片结果相一致。结论 MHCC-97H及Hep3B两种转移能力不同的肝癌细胞株的miRNAs存在差异表达,这些差异表达的miRNAs与肝细胞癌的发生发展、转移侵袭密切相关。

Aim To screen the differential microRNA(miRNA)expression profiles of different metastatic potential liver cancer cell lines,and predict miRNAs-regulated target genes and their functions.Methods Total RNA was extracted and the miRNA expression profiles were obtained by miRNA microarray chip hybridization.The miRNAs whose expression had significant difference were selected by analyzing the miRNA difference expression profiles of the two different metastatic potential liver cancer cell lines,namely MHCC-97H(high-metastasis)and Hep3B(non-metastasis),which were compared with normal hepatocytes L02 respectively.Moreover,we analyzed the miRNA differential expression profile between liver cancer cell lines MHCC-97H and Hep3B.The miRNAs were verified by qPCR and target genes were predicted by four softwares(TargetScan,miRanda,miRWalk,miRDB).To understand the biological functions of predicted target genes,bioinformatics analysis was performed.Results The miRNA microarray results showed that the expression of miR-192-5p and miR-215-5p significantly increased in liver cancer cell lines(MHCC-97H,Hep3B)when compared with normal hepatocytes L02,while miR-130a-3p and miR-196a-5p were significantly reduced;compared with Hep3B,the expression of miR-224-5p markedly increased in liver cancer cell line MHCC-97H,while miR-146a-5p,miR-483-3p and miR-200b-3p were significantly reduced.The results of qRT-PCR were consistent with chip results.Conclusion There are differences of miRNA expression profiles in different metastatic potential liver cancer cell lines MHCC-97H,Hep3B,and they may participate in regulating the development and invasion of hepatocellular carcinoma.