TITLE:
Inhibition of Hepatitis C Virus Genotype 1a Non-Structural Proteins by Small Interference RNA in Human Hepatoma Cell Lines
AUTHORS:
Imran Shahid, Waleed Hassan AlMalki, Shaia Saleh R. Almalki, Ismail Muhammad AlTurkestany, Hassan Ali AlGhamdi, Saleh Ali AlMenshawi
KEYWORDS:
Hepatitis C Virus, Non-Structural Proteins, Stable Cell Line, Anti-HCV Drugs, Short Interference RNA
JOURNAL NAME:
Pharmacology & Pharmacy,
Vol.6 No.11,
November
24,
2015
ABSTRACT: Hepatitis C virus (HCV) infection and associated liver diseases are still challenging and represent a significant health care burden around the world. Although, the treatment strategies have been improved by the development of novel direct-acting antivirals, but such therapeutic options are still expensive and beyond the financial range of the most infected individuals in developing or even in resource replete countries. It demands an urgent need to search novel and improved alternate treatment strategies to treat the infection. The present study was aimed to develop an in vitro stable cell culture system, persistently expressing HCV genotype 1a non-structural genes and to characterize the inhibitory effects of synthetic siRNAs (short interference RNA) directed against the most conserved regions of nonstructural genes in an in vitro cell culture model. The continuous expression of nonstructural genes for more than 30 days post transfection was detected by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis in stable human hepatoma cell line (Huh-7). The gene expression studies revealed significantly reduced gene expression of HCV nonstructural genes (i.e., NS2, NS4A and NS5A) both at mRNA and protein levels when treated against genome specific synthetic siRNAs in stable cell lines (51%, 47% and 54% respectively, p