Antiviral Study Of M1GS RNA Ribozyme Targeting Hepatitis B Virus C Gene Against HBV In HepG2.2.15 Cells

Objective: To evaluate the effect of M1GS RNA ribozyme targeting Hepatitis B virus (HBV ) c mRNA on the gene expression and replication of Hepatitis B virus (HBV) in HepG2.2.15 cells.Methods: Amplifying ,extracting and restriction enzyme analysis were used to identify the recombinant plasmid named pEGFP-GS expressing M1GS RNA ribozyme and pEGFP-C1 vector. Preparing eukaryotic plasmids for the purpose of transfection. The plasmids were then transfected into HepG2.2.15 by Lipofectamine TM 2000. The quantity of viral antigens were measured using a method of enzyme linked immunosorbentassay (ELISA). The levels of viral mRNA were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The amounts of HBV DNA secreted into the culture media were measured by quantitative real-time PCR. MTT assay was performed to determine the effect of the Ml RNA ribozyme on the proliferation ofHepG2.2.15 cells.Results: Vector-based M1GS RNA ribozyme potently reduced HBeAg expression in cellculture, the inhibition rate is 31.58%. The transcriptional level of HBV C mRNA wasgreatly decreased by up to 32.5%. However, there were no significant differences on theamounts of the HBV DNA between any two groups. The expression of the M1 RNAribozyme had no effect on proliferation of HepG2.2.15 cells.Conclusion: The M1GS RNA ribozyme could inhibit the expression of HBV c gene inHepG2.2.15 cells. It may be considered as a potential approach for anti-HBV genetherapy.