Expression and cytotoxic effect of recombinant Newcastle Disease Virus (rNDV) vector expressing enhanced green fluorescent gene in JHH5 cell line

Abstract

Introduction and Aim: In medicine, the term ‘cancer’ is used to describe a group of disorders defined by uncontrolled cell growth and the ability to invade and metastasize to other organs. As a result of the body's ability to fight conventional cancer treatments, it is essential to look for alternative therapeutic avenues. Recent years have seen a rise in the application of reverse genetics techniques including the Newcastle disease virus as an oncolytic agent in tumor models of immunocompetent malignancy. In preclinical studies, recombinant NDV (rNDV-GFP) that expresses foreign genes was shown to be more effective in cancer therapy. The goal of this study was to examine the in vitro anticancer effects of GFP-expressing, genetically modified Newcastle disease virus (rClone3-GFP) strains on the JHH5 hepatocellular carcinoma cell line. Materials and Methods: To generate a recombinant NDV, the GFP gene was inserted into the genome of an NDV strain (NDVClone30) by reverse genetics technology, where it now resides in the region of the genome occupied by the F and HN genes (rNDV-GFP). Using the MTT assay, we evaluated rNDV-GFP for its oncolytic potential against JHH5 cancer cells. Cytopathic effects were analyzed using fluorescent and light microscopy. Results: We observed that 96 hours post infection, the recombinant virus was capable of inducing tumor cell death in a time-dependent way. Conclusion: Recombinant NDV strains expressing GFP, demonstrated good results in inhibiting tumor growth. Our results pave the door for the application of recombinant NDV as a viral vector for the treatment of liver cancer.