Altered expression profiles of microRNAs in a stable hepatitis B virus-expressing cell line
Altered expression profiles of microRNAs in a stable hepatitis B virus-expressing cell line
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稳定的乙型肝炎病毒表达细胞系中 microRNA 的表达谱发生改变
DOI：
10.3760/cma.j.issn.0366-6999.2009.01.003
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发表时间：
2009-01-05
影响因子：
6.1
通讯作者：
Xu Dong-ping
中科院分区：
文献类型：
--
作者：
Liu Yan;Zhao Jian-Jun;Xu Dong-ping
MicroRNAs （miRNAs）是高度保守的18-25个核苷酸（nt）的小非编码rna，介导转录后基因调控。乙型肝炎病毒（HBV）可引起急性或慢性乙型肝炎，是肝硬化和肝细胞癌的高危因素。一些哺乳动物病毒已被证明可以调节宿主细胞mirna的表达。然而，HBV与宿主细胞mirna之间的相互作用在很大程度上是未知的。方法采用miRNA芯片和Northern blotting分析方法，比较稳定表达hbv的细胞株HepG2.2.15及其亲本细胞株HepG2的细胞miRNA表达谱。采用mRNA微阵列分析和miRanda程序预测miRNA靶标。流式细胞术进一步检测人白细胞抗原(HLA)-A的表达。结果18种mirna在两种细胞系中表达差异。其中，在HepG2.2.15细胞中上调11个，下调7个。Northern blotting分析证实miR-181a、miR-181b、miR-200b和miR-146a表达上调，miR-15a表达下调，与芯片分析结果一致。此外，一些推测的miRNA靶点被预测并证实与mRNA表达相关。HLA-A基因的3'-UTR与miR-181a有一个部分互补位点，miR-181a可能下调HLA-A的表达。结论HBV复制调节宿主细胞mirna的表达，可能在HBV相关肝病的发病机制中发挥作用。
Background MicroRNAs (miRNAs) are highly conserved small non-coding RNAs of 18-25 nucleotides (nt) that mediate post-transcriptional gene regulation. Hepatitis B virus (HBV) can cause either acute or chronic hepatitis B, and is a high risk factor for liver cirrhosis and hepatocellular carcinoma. Some mammalian viruses have been shown to modulate the expression of host cellular miRNAs. However, interactions between the HBV and the host cellular miRNAs are largely unknown.Methods miRNA microarray and Northern blotting analysis were used to compare the expression profile of cellular miRNAs of a stable HBV-expressing cell line HepG2.2.15 and its parent cell line HepG2. mRNA microarray assay and the miRanda program were used to predict the miRNA targets. A flow cytometric assay was further used to investigate the expression of human leukocyte antigen (HLA)-A.Results Eighteen miRNAs were differentially expressed between the two cell lines. Among them, eleven were up-regulated and seven were down-regulated in HepG2.2.15 cells. Northern blotting analysis confirmed that the expression of miR-181a, miR-181b, miR-200b and miR-146a were up-regulated and the expression of miR-15a was down-regulated, which was in consistent with the results of the microarray analysis. Furthermore, some putative miRNA targets were predicted and verified to be linked with mRNA expression. The 3'-UTR of HLA-A gene had one partially complementary site for miR-181a and miR-181a might down-regulate the expression of HLA-A.Conclusion HBV replication modulates the expression of host cellular miRNAs, which may play a role in the pathogenesis of HBV-related liver diseases.