Effects of MicroRNA-133b on epithelial-mesenchymal transition of human small airway epithelial cells induced by cigarette smoke extracts / 中华老年医学杂志

ABSTRACT

Objective:To investigate the effects of microRNA(miR)-133b on epithelial-mesenchymal transition(EM)of human small airway epithelial cells induced by cigarette smoke extracts(CSE)and its regulatory mechanisms.Methods:The miR-expression profiles with microarray in airway epithelial cells of patients with chronic obstructive pulmonary disease were searched in the Gene Expression Omnibus(GEO)database, and the differentially expressed miRs were searched and verified by a real-time fluorescence quantitative method(qRT-PCR). Human small airway epithelial cells(HSAEpiC)were divided into the control group, the CSE group, the CSE+ miR-133b inhibitor transfection group(inhibitor group)and the CSE+ miR-133b inhibitor negative control transfection group(inhibitor control group)according to different intervention methods.Levels of miR-133b and mRNA levels of transforming growth factor(TGF)-β1, Smad2, E-cadherin and vimentin were detected by RT-PCR; Protein levels of E-cadherin and vimentin were detected by enzyme-linked immunosorbent assays(ELISA)and Western blotting.Results:Nine differentially expressed miRs were found in GSE53519, with miR-133b showing the most significant differential in thee HSAEpiC cell model after verification.CSE induced morphological changes in HSAEpiC cells, and miR-133b inhibitors could partially reverse the morphological changes in cell mode.mRNA and protein expressions of E-cadherin were decreased and expression of Vimentin mRNA and protein were increared in CSE induced HSAEpiC cells( F=9.09、12.35、7.57、101.87, P=0.015、0.007、0.023、0.000); miR-133b inhibitors partally reversed the mRNA and protein expressions of E-cadherin and Vimentin( F=40.59、27.74、15.87、20.42, P=0.000、0.001、0.004、0.002). CSE induced incresed expression of TGF-β1 mRNA and Smad mRNA in HSAEpiC cells, and miR-133b inhibitors partially reversed the changes in TGF-β1 mRNA and Smad mRNA( F=17.25、64.15, P=0.003、0.000). Conclusions:miR-133b may regulate CSE-related HSAEpiC cell EMT through the TGF-β1/Smad pathway.