The Effect Of TGF-β1 On The Airway Remodeling And The Expression Of Six1in Human Bronchial Epithelial Cells

Objective: Investigate the relationship between TGF-β1 and the Epithelial-Mesenchymal Transition in human bronchial epithelial cells(16HBE),study the effect of TGF-β1 on the expression of Six1.Methods: Culture human bronchial epithelial cells(16HBE)in vitro,and build the model of Epithelial-Mesenchymal Transition.The morphological changes of cells was observed by inverted microscope;The expression of Six1,E-cadherin,vimentin,α-SMA and fibronectin m RNA was detected by Real-time PCR;and the expression of Six1,E-cadherin,vimentin,α-SMA,fibronectin protein was detected by Westernblotting.Results: 1.The inverted microscope showed: Cells in control group showed cobblestone shaped、tight junctions between epithelial cells;Cells in group A showed little obvious change;Cells in group B showed some cells hypertrophed,cell gap increased;Cells in group C showed fusiform shape,cell gap increased and even loss the typical paving like change.2.RT-PCR analysis: Cells in control group can normally express the m RNA of E-cadherin,vimentin,α-SMA,fibronectin Six1 gene.In Group A,the expression of Ecadherin and α-SMA m RNA was decreased(P=0.862 、 0.566 > 0.05),Six1 m RNA was increased(P=0.102>0.05),the difference has no statistical significance;the expression of vimentin,fibronectin m RNA was increased significantly(P=0.003、0.006<0.01),there has significantly difference.In Group B,the expression of E-cadherin m RNA was reduced(P=0.042),vimentin,fibronectin,α-SMA and Six1 gene m RNA were significantly increased compared with the cells in control group(P=0.000、0.001、0.031、0.023,< 0.05),the difference was statistically significant.The expression of E-cadherin m RNA in Group C was decreased significantly(P=0.004 < 0.01),vimentin,fibronectin,α-SMA and Six1 m RNA was significantly increased(P=0.004 、 0.003 、 0.023 、 0.004<0.05),the difference was statistically significant.3.Westernblot analysis: Cells in control group can normally express the protein of E-cadherin,vimentin,α-SMA,fibronectin Six1 protein.In Group A,the expression of Ecadherin protein decreased(P=1.023),α-SMA,fibronectin and vimentin protein increased(P= 0.789,0.542,0.624,> 0.05),but the differences were not statistically significant,the expression of Six1 protein increased(P=0.028 < 0.05),he difference was statistically significant;In Group B,the expression of E-cadherin protein was reduced(P=0.011 < 0.05),α-SMA,fibronectin,vimentin and Six1 protein expression were significantly increased(P =0.002,0.001,0.004,0.009 < 0.01),there has significantly difference.The expression of E-cadherin protein in Group C was decreased significantly((P=0.009 < 0.01),the expression ofα-SMA,vimentin and Six1 protein increased significantly(P = 0.008,0 3,0.007,< 0.01),the expression of fibronectin protein was significantly increased(P = 0.05),and the longer time TGF-β1 stimulated,the higer of Six1 protein was expressed,the difference was statistically significant between every groups(F=33.50).Conclusion: TGF-β1 may induce Epithelial-Mesenchymal Transition in human bronchial epithelial cells by up regulation the expression of Six1,which is involved in the airway remodeling of asthma.