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.
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Project supported by the National Natural Science Foundation of China (Nos. 30471641 and 30872389) and the Natural Science Foundation of Zhejiang Province, China (No. Y207088)
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Shou, Zf., Zhou, Q., Cai, Jr. et al. PI-3 kinase pathway can mediate the effect of TGF-β1 in inducing the expression of SHARP-2 in LLC-PK1 cells. J. Zhejiang Univ. Sci. B 10, 702–706 (2009). https://doi.org/10.1631/jzus.B0920066
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DOI: https://doi.org/10.1631/jzus.B0920066
Key words
- Transforming growth factor (TGF)-β1
- Split- and hairy-related protein-2 (SHARP-2)
- Phosphoinositide 3-kinases (PI-3) kinase pathway
- LY294002