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标题： HOXA13 exerts a beneficial effect in albumin-induced epithelial-mesenchymal transition via the glucocorticoid receptor signaling pathway in human renal tubular epithelial cells

讲者：彭力

单位：中南大学湘雅二医院

关键词：

Homeobox A13 Epithelial-to-mesenchymal transition glucocorticoid receptor

播放： 1646

论文摘要： Objective To investigate the role of HOXA13 in HSA-induced EMT in HKC cells and the potential mechanism of the glucocorticoid receptor (GR) signaling pathway.
Methods HKC were treated with various amounts of HSA( 0，1，5，10，20，30mg/mL) for 48 hours or 20mg/mL HSA for different time periods ( 0，12，24，48，72hours)．The mRNA and protein expressions of CK，Vimentin and HOXA13 in HKC were detected by Real-time PCR，Western Blot and Immunohistochemical staining．Plasmid of HOXA13 was transfected into HKC with Lipofectamine 2000 and then treated with 20mg/mL of HSA. Real-time PCR and Western Blot were applied to detect the protein expression of CK，Vimentin and HOXA13 in HKC. Then the mRNA and protein expression of GR，CBP was detected by Real-time PCR and Western Blot in HSA (20mg/mL) -induced HKC 48h；Plasmid of HOXA13 was transfected into HKC and treated with 20mg/mL of HSA，and then cells were cultured in the present and absent of 1μg /mL CBP，Western Blot was applied to detect the protein expression of GR、CBP、CK and Vimentin in HKC.
Results The mRNA and protein expressions of CK in HKC downregulated while the mRNA and protein expressions of Vimentin upregulated in a dose-dependent and time-dependent manner in response to HSA treatment．Meanwhile，the mRNA and protein expressions of HOXA13 also decreased in a dose-dependent and time-dependent manner when detected with real-time PCR and Western Blot. The Immunohistochemical staining showed that HOXA13 expressed in nucleus and cytoplasm，and especial positively in the nucleus in normal HKC，however，staining of HOXA13 protein decreased especially in the nuclear region after HSA 20 mg/mL for 48 hours．When overloaded with 20mg/mL HSA，the level of HOXA13 expression positively correlated with CK but negatively correlated with Vimentin expression (P<0.05). Protein expression of HOXA13 could be effectively and specifically upregulated by Lipofectamine 2000 when detected by Western Blot．Overexpression of HOXA13 significantly induced CK expression and suppressed the protein expression of Vimentin．The mRNA and protein expressions of GR in HKC
downregulated while the mRNA and protein expressions of CBP upregulated in response to HSA treatment．Overexpression of HOXA13 significantly induced the protein expression of GR and suppressed CBP expression；but Western Blot showed that CBP could alleviate the upregulation of GR protein by overexpression of HOXA13，at the same time，the expression of CK was down-regulated and the expression of Vimentin was up-regulated.
Conclusion HOXA13 is involved in HSA?induced EMT in HKC cells and upregulation of HOXA13 exerts a beneficial effect in EMT, which may be associated with the GR signaling pathway.