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Focal adhesion kinase mediates TGF-β1-induced renal tubular epithelial-to-mesenchymal transition in vitro

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Abstract

Focal adhesion kinase (FAK) is a non-receptor tyrosine kinase which participates in many important cellular processes such as cell adhesion and migration. However, the role of FAK in renal tubular epithelial-to-mesenchymal transition (EMT) is still unknown. FAK was knocked down by transfection of specific small interfering RNA (siRNA) in cultured HK-2 cells, then the cells were stimulated with transforming growth factor-beta 1 (TGF-β1). The expression of FAK, α-smooth muscle actin (α-SMA),E-cadherin, Akt, matrix metallopeptidase-9 (MMP-9),tissue inhibitor of metalloproteinase-1 (TIMP-1), and collagen IV were detected by RT-PCR, Western blot and immunofluorescence methods, respectively. Cell migration was determined by transwell assay. The results suggest that the expression of FAK was up-regulated in HK-2 cells when incubated with TGF-β1(10 μg/l), which was accompanied by reduced expression of E-cadherin and increased expression of α-SMA. All these changes were restored by FAK siRNA. Akt phosphorylation was induced by the treatment with TGF-β1, which was blocked by FAK siRNA. TGF-β1-induced down-regulation of E-cadherin was recovered by a PI3K/Akt inhibitor, LY294002, without affecting the expression of FAK. Functionally, TGF-β1 induced an increase in MMP-9 expression, as well as decreased expression of TIMP-1 and collagen IV, which were all restored by the FAK siRNA transfection. In addition, FAK siRNA significantly reduced TGF-β1-induced cells migration. In conclusion, FAK may play a crucial role in mediating TGF-β1-induced EMT through the activation of Akt pathway.

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Acknowledgments

This study was supported by grants from National Natural Science Foundation of China (No.30871174).

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Authors and Affiliations

  1. Department of Nephrology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China

    Bingqing Deng, Xiao Yang, Jianshe Liu, Fangfang He, Zhonghua Zhu & Chun Zhang

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  1. Bingqing Deng
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  2. Xiao Yang
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  3. Jianshe Liu
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  5. Zhonghua Zhu
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Correspondence to Zhonghua Zhu or Chun Zhang.

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Deng, B., Yang, X., Liu, J. et al. Focal adhesion kinase mediates TGF-β1-induced renal tubular epithelial-to-mesenchymal transition in vitro. Mol Cell Biochem 340, 21–29 (2010). https://doi.org/10.1007/s11010-010-0396-7

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  • DOI: https://doi.org/10.1007/s11010-010-0396-7

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