HuH-7肝きも癌がん細胞さいぼうにおいて、SHARP-2遺伝子いでんしの発現はつげんは、トランスフォーミング成長せいちょう因子いんしβべーたにより様々さまざまな経路けいろを介かいして誘導ゆうどうされる

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  • Gene expression of SHARP-2 is induced by transforming growth factor beta via multiple pathways in HuH-7 hepatoma cells

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Transforming growth factor-β (TGF-β) inhibits cell proliferation and promotes cell differentiation. In human HuH-7 hepatocellular carcinoma cells, TGF-β reduces the expression of the α-fetoprotein (AFP) gene, a marker molecule for hepatocellular carcinoma cells. In this case, the amount of AT motif-binding factor-1 (ATBF1) mRNA, which encodes a transcriptional repressor of the AFP gene, was not changed. This suggests the involvement of factors that are induced by TGF-β and interact with ATBF1. The enhancer of split- and hairy-related protein-2 (SHARP-2) is a basic helix-loop-helix transcriptional repressor and an interacting protein of ATBF1. In this study, we examined whether TGF-β regulates the expression of the SHARP-2 gene in HuH-7 cells. TGF-β induced the SHARP-2 mRNA level within 1 hr. Next, to explore the signaling pathway, HuH-7 cells were pretreated with various inhibitors and then treated with TGF-β. Inhibitors of TGF-β type I receptor, phosphoinositide 3-kinase, protein kinase C, Jun N-terminal kinase, mitogen-activated protein kinase, and RNA polymerase II completely or partially diminished the induction of the SHARP-2 mRNA level by TGF-β. Finally, a reporter plasmid containing from -3,700 to +265 region of the SHARP-2 gene were transfected into HuH-7 cells and treated with TGF-β, but no change in promoter activity was observed. Thus, we concluded that TGF-β increased the level of SHARP-2 mRNA via multiple pathways at the transcription level of the SHARP-2 gene and that SHARP-2 is a candidate protein mediating TGF-β-repression of the AFP gene.

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