The microRNA gene expression profiling in HepG2 cells transfected with full-long hepatitis B virus genome
- VernacularTitle:乙型肝炎病毒基因组转染HepG2细胞的microRNA表达研究
- Author:
Yan LIU
;
Liang ZHANG
;
Jiuzeng DAI
- Publication Type:Journal Article
- Keywords:
hepatitis B virus;
microRNAs;
gene expression
- From:
Medical Journal of Chinese People's Liberation Army
1983;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective MicroRNA(miRNA)has recently been suggested to play a role in certain virus infections.The present study is to investigate if miRNA is associated with hepatitis B virus(HBV)activity by detecting differential expression of miRNA between human hepatoblastoma cell line HepG2.2.15 transfected with full-long HBV genome and its parent cell line HepG2.Methods Total RNA were extracted from HepG2.2.15 cells and control HepG2 cells,respectively.miRNAs were then isolated from the total RNA.Mammalian miRNA microarrays containing 509 miRNA genes were employed to analyze the differentially expressed miRNAs between the two groups.miRNAs were considered to be up-or down-regulated when the fluorescent intensity ratio between the two groups was over 4-fold and global false positive is zero using SAM program.Validation of microarray results was carried out by real-time quantitative RT-PCR(qRT-PCR).Results Compared with those of control HepG2 cells,a total of 27 miRNAs were differentially expressed(5.3% of all probes),among which 7 were up-regulated and 20 were down-regulated in HepG2.2.15 cells.qRT-PCR verified that miR-181d expression was 16-fold up-regulated and miR-15a was 9-fold down-regulated in HepG2.2.15 cells,which was in agreement with the results of the microarray analysis.Conclusion The findings suggest that there are HBV replication-associated miRNAs in HBV genome-transfected HepG2.2.15 cells.These up-and down-regulated miRNAs may be involved in the life cycle of HBV replication.The knowledge is helpful for further study to discover new molecular targets for anti-HBV therapy.
