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 hep...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-181 a 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.展开更多
Recently the Chinese Ministry of Health declared that 93 million people wer.e chronically infected with hepatitis B virus （HBV） in China. Four nucleotide analogues （NAs） are currently approved for the treatment of...Recently the Chinese Ministry of Health declared that 93 million people wer.e chronically infected with hepatitis B virus （HBV） in China. Four nucleotide analogues （NAs） are currently approved for the treatment of HBV infection, i.e., lamivudine （LAM）, adefovir （ADV）, entecavir （ETV）, and telbivudine （L-dT）. In contrast to therapeutic benefits, prolonged use of NAs increases the emergence of drug-resistant mutations in the HBV reverse transcriptase （RT） domain, leading to a progressive accumulation of NAs-resistant patients. Especially, LAM-refractory patients are prone to develop cross-resistance to other NAs. Novel anti-HBV drugs that are able to deal with the cross-resistance are needed in the clinic. Tenofovir （TDF）, which was recently approved in Europe and United States of America, has potential to be such a drug. Clinical trials have proven its high potency for inhibiting HBV replication in HBV-infected patients,展开更多
基金Supported by grants from the National Key Basic Research Programof China (2007CB512803)Key Project of Beijing Natural Science Foundation (7091006)National 11th Five-Year Special Grand Project for Infectious Diseases(2008ZX10002-005-6,2008ZX10002-011)
基金This study was supported by the grants from the National Natural Science Foundation of China （No. 30700714） and the Major State Basic Research Development Program of China （973 Program, No. 2007CB512803）.Acknowledgement： We thank Dr. ZHANG Liang from Beijing Capital Bio Corporation of China for his helpful advice and comments.
文摘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-181 a 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.
基金The study was supported by the grants from the National Key Basic Research Developing Project （No. 2007CB512803）, the Beijing Science and Technology Project （No. Z07050700690702）, and partly by the Beijing Natural Science Foundation （No. 7091006）.Acknowledgement： The authors gratefully acknowledge experimental participation of technicians at Viral Hepatitis Research Laboratory as follows： DAI Jiu-zeng, LI Le, YAO Zeng-tao.
文摘Recently the Chinese Ministry of Health declared that 93 million people wer.e chronically infected with hepatitis B virus （HBV） in China. Four nucleotide analogues （NAs） are currently approved for the treatment of HBV infection, i.e., lamivudine （LAM）, adefovir （ADV）, entecavir （ETV）, and telbivudine （L-dT）. In contrast to therapeutic benefits, prolonged use of NAs increases the emergence of drug-resistant mutations in the HBV reverse transcriptase （RT） domain, leading to a progressive accumulation of NAs-resistant patients. Especially, LAM-refractory patients are prone to develop cross-resistance to other NAs. Novel anti-HBV drugs that are able to deal with the cross-resistance are needed in the clinic. Tenofovir （TDF）, which was recently approved in Europe and United States of America, has potential to be such a drug. Clinical trials have proven its high potency for inhibiting HBV replication in HBV-infected patients,
