Hepatitis B virus X protein induces expression changes of miR-۲۱, miR-۲۲, miR-۱۲۲, miR-۱۳۲, and miR-۲۲۲ in Huh-۷ cell line

Background: Hepatocellular carcinoma (HCC) is recognized as one of the most deadly malignant cancers which ranks third among all annual cancer mortality rates. The hepatitis B virus (HBV) X protein (HBX) is known to play a key role in HCC. The HBX may alter the expression of multiple microRNAs (miRs), which are important in hepatocarcinogenesis. This study aimed to investigate the importance of HBX protein in mir۲۱, mir۲۲, mi۱۲۲, mir۱۳۲, and mir۲۲۲ expression.Material and Methods: In the present study, a recombinant vector, pcDNA۳.۱+ expressing HBx was developed. The Huh-۷ cell line was transfected with HBx-pcDNA۳.۱+ recombinant plasmid. Real-Time Polymerase Chain Reaction was used to evaluate mir۲۱, mir۲۲, mi۱۲۲, mir۱۳۲, and mir۲۲۲ expression in the cell line.Results: It was found that expression of miR-۲۱ and miR-۲۲۲ was upregulated at all points of time since HBx transfection. The expression of miR-۲۱ reached to ۴.۲۴-fold after ۷۲ hours post-transfection. The miR-۲۲ had a ۷.۶۹-fold downregulation after ۲۴ hours. The miR-۱۲۲ had a significant downregulation after ۴۸ (۱۰-fold) hours. The miR-۱۳۲ expression reached its lowest rate at ۱۲ hours after HBx-transfection (۸.۳۳-fold). The miR-۲۲۲ expression was upregulated in transfected cells but was not significantly different (۱.۱۸ to ۲.۴۵-fold).Conclusion: Significant downregulation of miR-۲۲, miR-۱۲۲ and miR-۱۳۲ implicate their inhibitory roles in the progression of HBV-associated HCC. The expression of these microRNAs could be used as a prognosis of the progression of HBV-associated liver disease. Also, in future studies, miR-۲۱ and miR-۲۲ can be used as a target in the development of therapeutic agents.

Keywords:

Hepatitis B virus x protein , HBx , Micro RNA , miR-۲۱