Hepatocyte Growth Factor and Betacellulin Gene Expression for Treating Diabetes: In vitro analysis

Abstract

Background: Diabetes mellitus is a fast spreading metabolic disease in both developed and developing countries owing to genetic predisposition, lifestyle and or environmental conditions. Recombinant insulin used for managing diabetes has been approved in 1985, but till now there has been no cure. Insulin cannot help the patients who become resistant to it. Islet transplantation has been developed in 2001 and can become a possible cure for Diabetes. Gene therapy can be beneficial in ameliorating the functionality and survival of islets post-transplantation. Plasmid vectors pcDNA3.1 BTC and pcDNA3.1 HGF can help with proliferation and protection of islet cells from apoptosis. Methods: Plasmid vectors encoding genes Betacellulin (BTC) and Hepatocyte growth factor (HGF) were constructed and the gene expression, caspase -3 and cell viability assays were done to assess the activity of these vectors. Results: pcDNA3.1 BTC and pcDNA3.1 HGF showed time dependent increase in expression of BTC and HGF following transfection in rat insulinoma (RIN-5mF) cells by forming complexes with lipofectamine-3000. There was no toxicity associated with the use of lipofectamine-3000 and the expression of BTC and HGF seemed to protect them from cytokine mediated apoptosis. Conclusion: The use of viral vectors is fraught with challenges especially with regards to their safety. Hence, we used plasmid vectors to express therapeutic genes Betacellulin and Hepatocyte growth factor and the use of these vectors on RIN-m5F cells showed that they could protect them from apoptosis mediated cell death and can enhance their function and survival.