Altered expression profiles of microRNAs in a stable hepatitis B virus-expressing cell line.
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.


CONCLUSIONS
HBV replication modulates the expression of host cellular miRNAs, which may play a role in the pathogenesis of HBV-related liver diseases.
Liu
Yan
Y
Viral Hepatitis Research Laboratory, Institute of Infectious Diseases, Beijing Hospital, Beijing, China.
Zhao
Jian-Jun
JJ
Wang
Chun-mei
CM
Li
Mian-yang
MY
Han
Ping
P
Wang
Lin
L
Cheng
Yong-qian
YQ
Zoulim
Fabien
F
Ma
Xu
X
Xu
Dong-ping
DP
D015152:Blotting, Northern; D045744:Cell Line, Tumor / Q000378:metabolism* / Q000821:virology*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D015234:HLA-A Antigens / Q000378:metabolism*; D006515:Hepatitis B virus / Q000254:growth & development* / Q000502:physiology; D006801:Humans; D035683:MicroRNAs / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis
D015234:HLA-A Antigens; D035683:MicroRNAs