Cloning and characterization of a transforming growth factor beta 1-induced anti-apoptotic adhesion protein TIF2

Transforming growth factor-beta (TGF-beta) antagonizes the cytotoxic function of tumor necrosis factor (TNF). By differential display and library screening, we isolated a murine TIF2 (TGF-beta-induced factor 2) cDNA, encoding a putative 15-kDa membrane adhesion protein, which possesses an RGD sequence at the extracellular region. When TNF-sensitive murine L929 fibroblasts were stably transfected with TIF2 cDNA, these cells significantly resisted TNF killing. In contrast, L929 cells, which stably expressed the TIF2 antisense mRNA, acquired enhanced TNF susceptibility. Calculated EC50 values, i.e., the amount of TNF needed for killing 50% cells, are 10, 55, and 1.5 ng/ml, respectively, for vector control, sense transfectant, and antisense transfectant. TGF-beta 1 rapidly induces TIF2 gene expression (approximately 1 hr), which correlates with time-related acquisition of TNF-resistance in TGF-beta 1-treated L929 cells. Notably, TIF2 gene expression is markedly increased in human breast cancer and lymphoid leukemia cells, compared to normal human cells, suggesting its potential role in cancer development. Together, the anti-apoptotic function of TIF2 is responsible in part for TGF-beta-mediated protection of L929 cells against TNF cytotoxicity.