Hepatocyte growth-promoting factor partially reverses monocyte chemotatic protein-1-and aristolochic acidI-induced epithelial-to-mesenchymal transition of human kidney epithelial cells
2010
Objective To observe the influence of hepatocyte growth-promoting factor ( pHGF) on monocyte chemotatic protein-1-(MCP-1) and aristolochic acid Ⅰ( AAⅠ)-induced epithelial-to-mesenchymal transition (EMT) and apoptosis of human kidney epithelial cell line( HKC). Methods The HKC cells were randomly divided into blank control group ( control groups) ,epithelial-to-mesenchymal transition model group ( model group) ,and pHGF inhibition group ( pHGF groups) with pHGF at different concentrations ( 0. 15,1. 5,15,150,and 1 500 ng/ml). The EMT model was established by exposing HKC cells to MCP -1( 0. 1 μg/ ml) and AAⅠ( 10 μg/ml). Cells in the pHGF groups were the model cells treated with different concentrations of pHGF. Cells in the control group were cultured routinely. WST -8 method and flow cytometry were used to observe the proliferation and apoptosis of HKC cells,respectively. The mRNA expression of α -smooth muscle actin( α-SMA) was determined by reverse transcriptase-polymerase chain reaction( RT-PCR) ,and the expression of α-SMA,fibronectin ( FN) ,and transforming growth factor-β1( TGF-β1) in HKC cells were assessed by indirect enzyme immunohistochemistry. Results The cell inhibitory rate,apoptotic rate,and expression of α-SMA mRNA were significantly increased in the model group and pHGF groups compared with those in the control group ( P 0. 01) ,indicating the successful establishment of EMT model. Compared with the model group,pHGF at 150 ng/ml,but not at other concentrations,significantly decreased the inhibition rate of HKC cells( P 0. 01). The apoptotic rate of HKC cells in all the pHGF groups were significantly lower than that in the model group ( P 0. 01). pHGF at 150 ng/ml also greatly decreased the expression of α -SMA mRNA,and significantly down-regulated the expression of α-SMA,TGF-β1,and FN protein. Conclusion pHGF at 150 ng/ml can partly reverse MCP-1-and AA Ⅰ-induced HKC cell growth inhibition,apoptosis,and EMT.
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