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ZJUS-B
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology)  2009, Vol. 10 Issue (9): 702-706    DOI: 10.1631/jzus.B0920066
Biomedicine     
PI-3 kinase pathway can mediate the effect of TGF-β1 in inducing the expression of SHARP-2 in LLC-PK1 cells
Zhang-fei SHOU, Qin ZHOU, Jie-ru CAI, Jiang-hua CHEN, Kazuya YAMADA, Kaoru MIYAMOTO
Kidney Disease Center, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China; Key Laboratory of Multi-organ Combined Transplantation, Ministry of Health, Hangzhou 310003, China; Department of Biochemistry, Faculty of Medical Sciences, University of Fukui, Eiheiji-cho, Fukui 910-1193, Japan
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Abstract  We aim to investigate the effect of transforming growth factor (TGF)-β1 on the expression of enhancer of split- and hairy-related protein-2 (SHARP-2) messenger RNA (mRNA) and its signaling pathway. In this study, several cell lines including LLC-PK1 (a porcine kidney tubular epithelial cell line), MDCK (Madin-Darby canine kidney) and CTLL-2 (cytotoxic T-lymphocyte line) were treated with recombinant human TGF-β1, and a series of experiments were carried out, involving Northern blot analysis of total RNA from these cells. Further, several specific chemical inhibitors were applied before TGF-β1 treatment to probe the signaling pathway. The results showed that TGF-β1 can significantly up-regulate SHARP-2 mRNA expression in the LLC-PK1 cell line. The peak level of induction was found 2 h after TGF-β1 stimulation. While one phosphoinositide 3-kinases (PI-3) kinase inhibitor, LY294002, completely blocked the effect of TGF-β1 on SHARP-2 mRNA expression in LLC-PK1 cells at a low concentration, other inhibitors, including PD98059, staurosporine, AG490, wortmannin, okadaic acid and rapamycin, had no effect. The effect of LY294002 was dose-dependent. We conclude that, in LLC-PK1 cells at least, TGF-β1 can effectively induce the SHARP-2 mRNA expression and that the PI-3 kinase pathway can mediate this effect.

Key words： Transforming growth factor (TGF)-β1      Split- and hairy-related protein-2 (SHARP-2)      Phosphoinositide 3-kinases (PI-3) kinase pathway      LY294002     
Received: 03 March 2009     
CLC:  Q26  
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Zhang-fei SHOU
Qin ZHOU
Jie-ru CAI
Jiang-hua CHEN
Kazuya YAMADA
Kaoru MIYAMOTO
Cite this article:

Zhang-fei SHOU, Qin ZHOU, Jie-ru CAI, Jiang-hua CHEN, Kazuya YAMADA, Kaoru MIYAMOTO. PI-3 kinase pathway can mediate the effect of TGF-β1 in inducing the expression of SHARP-2 in LLC-PK1 cells. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2009, 10(9): 702-706.

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http://www.zjujournals.com/xueshu/zjus-b/10.1631/jzus.B0920066     OR     http://www.zjujournals.com/xueshu/zjus-b/Y2009/V10/I9/702

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