Expression of paxillin in human kidney proximal tubular epithelial cell line undergoing tubular epithelial to mesenchymal transition induced by TGFβ1 and its significance

Abstract

Objective To investigate the expression of paxillin （Pax） in human kidney proximal tubular epithelial cell line （HKC cells） undergoing tubular epithelial to mesenchymal transition （TEMT） induced by transforming growth factorβ1 （TGFβ1 ） and intervention effect of genistein （Gen） on TEMT. Methods HKC cells cultured in vitro were random divided into three groups ： Control （C） group, the HKC cells were cultured with free serum medium （FSM） , amounting to being cultured with FSM containing 0ng/ml TGFβ1 ; TGFβ1 -treated （T） groups, the HKC cells were incubated with FSM containing different concentrations of TGFβ1 （T5 group： containing 5 ng/ml TGFβ1 ; T10 group： containing 10ng/ml TGFβ1 ）. After treatment for 48 hours, the mRNA and protein expression of E-cadherin, alpha-smooth muscle （ct-SMA） and Pax in HKC cells was detected by reverse transcription polymerase chain reaction （RT-PCR） and immunohistochemical staining respectively. The expression of Pax protein was detected by western blot at the same time. Results （1）C group ： the mRNA and protein expression of E-cadherin was high expressed in HKC cells while the mRNA and protein expression of α-SMA was negative, and the mRNA and protein expression of Pax was elementary; （2）T groups： the statistically significant changes were observed after the HKC calls were stimulated with different concentrations of TGFβ1. The mRNA and protein expression of E-cadherin in HKC cells reduced dramatically in both groups versus C group （ P 〈 0. 01 ） , and the reduction was statistically significant in T10 group compared with T5 group （ P 〈 0. 01 ） while the mRNA and protein expression of α-SMA and Pax in HKC cells increased evidently in both groups versus C group （ P 〈 0. 01 ） and the increase was statistically significant in T10 group compared with T5 group （ P 〈 0. 01 ） following addition of TGFI31 ; （3）T ＋ G group ： The mRNA and protein expression of E-cadherin in HKC cells increased dramatically versus T10 group （ P 〈 0. 01 ）, and the increase was statistically significant compared with C group （ P 〈0. 01 ）, while the mRNA and protein expression of α-SMA and Pax in HKC cells decreased evidently versus T10 group （ P 〈 0. 01 ） and the decrease was statistically significant compared with versus C group （ P 〈 0. 01 ） following addition of Gen （50μmol/L） and TGFβ1 （ 10 ng/ml）. Conclusion TGF[31 could induce Pax expression during it triggered and mediated TEMT in HKC cells in a dose-dependent manner. Gen （50 μmol/L） could partly inhibit TEMT in HKC cells that was induced by TGFβ1 （ 10ng/ ml）, and the mRNA and protein expression of Pax in HKC cells was stimulated by TGFβ1. Key words: Cytoskeletal proteins; Transforming growth factor beta; Kidney tubulesiEpithelial cells; Isoflavones/PD