TGF-β1 induces activation ofHSC-T6 cells and epithelial-mesenchymal transition in rats
- VernacularTitle:TGF-β1诱导大鼠肝星状细胞系HSC-T6活化及上皮间质转换
- Author:
Lihui WANG
;
Bianqiao CHENG
;
Qi ZHU
;
Weiguo LIN
- Keywords:
TGF-β1;
hepatic stellate cells;
activation;
epithelial-mesenchymal transition
- From:
Basic & Clinical Medicine
2017;37(9):1257-1262
- CountryChina
- Language:Chinese
-
Abstract:
Objective To observe the effect of TGF-β1 on activation and epithelial mesenchymal transition(EMT) in rat hepatic stellate cell-T6.Methods Adopt the MTT method to screen the optimum concentration of TGF-β1 in vitro HSC-T6 cultured.After the HSC-T6 stimulation by TGF-β1 of 10 μg/L for 24 hours, the morphology of the cells was observed under inverted phase contrast microscope, the expression of F-actin which on behalf of cotoskeletal structure was detected by immunofluorescence staining;the expression of α-SMA and N-cadherin,vimentin,E-cadherin was measured by RT-qPCR;The changes of α-SMA,N-cadherin,vimentin and E-cadherin were assessed by Western blot after different concentrations (0,5 and 10 μg/L) of TGF-β1 interventing HSC-T6 for 24 h.Results The optimal cell survival rate was recorded when 10 μg/L TGF-β1 dealt withcells for 24 h.After HSC-T6 were treated with TGF-β1,cells stretched, pseudopodia increased and turn into stellate, cells connections were looser, so that represented a significantly activated state.F-actin filaments gathered to form stress and distributed along the long axis of the cells;The expression of α-SMA mRNA and vimentin mRNA in experimental group was significantly higher while E-cadherin mRNA was obviously lower than the control group(P<0.05).TGF-β1 made the protein expression of α-SMA and N-cadherin, vimentin in dose-dependent increased while E-cadherin was decreased.Conclusions TGF-β1 may induce activation and epithelial-mesenchymal transition of HSC-T6.
