Effect of RNA interference on the hepatitis C virus core expression in HepG2/C3A cells using genotype 4 isolates from Saudi patients

Abstract

Objective: To explore the effects of two different methodologies of RNA interference, namely small interfering RNA, and vector-based short hairpin RNA, on the expression levels of hepatitis C virus core RNA and protein of Saudi genotype 4 isolates. Study Design: An experimental study. Place and Duration of Study: Laboratories of the College of Medicine Research Center, King Saud University, Saudi Arabia, from January to December 2018. Methodology: Hepatitis C virus core small interfering RNA molecule and short hairpin RNA vector were designed against core region. Viral RNA expression was tested by RT-PCR; whereas, core protein was tested by flow cytometry and immunofluorescence. Results were statistically analysed by Chi-square analysis to calculate the p-value. Results: Both molecules caused a reduction in core RNA and protein expression in infected cells. The effect of 100-pmol of small interfering RNA was more evident. For the vector-based short hairpin RNA, inhibition of core RNA expression was quite evident after 96 hours (p = 0.007). The results of flow cytometry and immunofluorescence showed a decline in core protein expression. The most dramatic effect was observed with 100-pmol small interfering RNA treatment of cells for 24 and 48 hours, which resulted in 63.5% and 91.1% core RNA expression reduction, respectively. Conclusion: RNA interference of hepatitis C virus core gene efficiently stopped viral replication and offer a promising therapeutic alternative against virus infection.