埃兹蛋白在转化生长因子-β诱导的支气管上皮细胞间充质转化中的作用

The effects of ezrin on transforming growth factor beta mediated epithelial mesenchymal transition

目的探讨埃兹蛋白(ezrin)在支气管上皮细胞(16HBE)间充质转化(EMT)中的作用。方法建立转化生长因子-β(TGF-β)诱导的16HBE的EMT模型;RT-PCR和Western blot法检测16HBE中ezrin m RNA和蛋白表达;通过慢病毒sh RNA抑制ezrin表达,观察细胞形态,检测细胞EMT的生物标志分子上皮钙黏着蛋白(E-cadherin)、波形蛋白(vimentin)和α-平滑肌动蛋白(alpha smooth muscle actin,α-SMA)的m RNA和蛋白表达水平;加入外源性ezrin蛋白,细胞免疫荧光、RT-PCR及Western blot法检测其对EMT标志物的影响。结果 TGF-β能下调16HBE中ezrin的m RNA和蛋白表达。经ezrin敲减后,同TGF-β刺激表现相同,16HBE也由典型的铺路石样变为梭形,同时其E-cadherin表达下降,vimentin和α-SMA表达增加;加入重组蛋白ezrin后,16HBE的E-cadherin表达增多,vimentin和α-SMA表达下降。结论 TGF-β可能通过下调ezrin表达促进16HBE发生EMT。

ObjectiveTo investigate the effects of ezrin on transforming growth factor beta （TGF-β） mediated epithelial mesenchymal transition （EMT）. MethodsHuman bronchial epithelial 16HBE cells were stimulated by TGF-β to induce EMT. The mRNA and protein levels were analyzed by RT-PCR and Western blot. Lentivirus （LV）-Ezrin-shRNA was employed to investigate the effects of ezrin deficiency on cell morphology and expressions of EMT associated biomarkers including E-cadherin, vimentin, and alpha smooth muscle actin （α-SMA）. The effects of recombinant ezrin protein on 16HBE were also examined. ResultsThe expression of ezrin was down-regulated in 16HBE activated by TGF-β. Due to ezrin depletion, the cell morphology changed from a typical multilateral paving stone-like appearance to a mesenchymal-like fusiform appearance along with the decreased expression of epithelium biomarker E-cadherin and the increased mesenchymal cell markers, vimentin and α-SMA. Recombination protein ezrin increased the expression of E-cadherin whilst reducing vimentin and α-SMA. ConclusionTGF-β promotes EMT of 16HBE cells at least partly via inhibiting the expression of ezrin.