Suppressed cellular synthesis of transforming growth factor(TGF) β1 by stably expressed antisense TGF β1

Abstract

The transforming growth factor(TGF)β1, as a set of multifunctional cytokine, has diverse action on a particular target cell according to the cell type and its state of differentiation, the growth conditions. When its production is increased under pathological conditions by cancer cells or by stimulated fibroblasts, TGFβ1 suppresses immune system and aggravates fibrotic complications associated with chronic inflammatory conditions. So, it is necessary to develop a strategy to inhibit the expression of TGFβ1 as a therapeutic application. The specific inhibition of gene using oligonucleotides complementary to its mRNA has been applied in many fields of gene therapy. The advanced antisense strategy is the stable expression of antisense mRNA carried by viral construct. The purpose of this study is to examine the degree of translational block of TGFβ1 synthesis from target cells which were infected by retroviral antisense TGFβ1. In this study, murine cell lines such as JC, K7, K12 and B16F1 cells were infected by antisense mouse TGFβ1 MFG and the synthesized TGFβ1 in culture media was measured by sandwitch ELISA kit. The range of inhibited degree for the synthesis of TGFβ1 was from 30% to 80% according to the cell types and the length of antisense mRNA at 48 hours after retroviral infection. To apply the antisense inhibition of TGFβ1 in vivo, it is essential to maximize the degree of inhibition by combining both of antisense and ribozyme strategy and to construct antisense TGFβ1 retrovirus with target tissue specific promoter.