Study on Establishment of Epithelial-Mesenchymal Transition and Regulation of Mahuang-Gancao Couplet Medicines in 16HBE Cells

OBJECTIVE To establish the epithelial-mesenchymal transformation (EMT) in 16HBE cells and to investigate the effects of couplet medicines on EMT in 16HBE cells. METHODS Western blot was applied to detect the mark proteins expression in 16HBE cells treated with TGF-β1 or combined with serum starvation. The distribution of E-Cadherin and N-Cadherin were detected by immunofluorescence. The qPCR was used for detecting the mark mRNA levels. The effects of couplet medicines on the expression of EMT associated proteins were detected by Western blot. RESULTS TGF-β1 induced 16HBE cells type changed， from a cobblestone to a spindle. TGF-β1 increased the expression of N-Cadherin and α-SMA， but not affected the expression of E-Cadherin and ZO-1. Meanwhile， the protein and mRNA expression of N- Cadherin， α-SMA， Vimentin， Collagen Ⅰ were significantly increased in 16HBE cells treated with serum starvation and TGF-β1. At the same time， the protein and mRNA levels of E-Cadherin and ZO-1 were obviously decreased. The distribution of E-Cadherin was disorder and the epithelial structure was damaged. Compared with the stimulate group， the couplet medicines decreased the expression of N-Cadherin and α-SMA， while increased the expression of E-Cadherin. CONCLUSION 16HBE cells， stimulated with serum starvation and TGF-β1， undergo the EMT process， which can be applied to study the mechanism of EMT in asthma. The Mahuang-Gancao couplet medicines can inhibit EMT in 16HBE cells.