MiR-152对转化生长因子β1诱导的气管上皮细胞向间质细胞转化的影响

The effects of miR-152 on transforming growth factor-beta1-mediated epithelial-mesenchymal transition in human bronchial epithelial cells

目的:研究miR-152在转化生长因子β1(TGF-β1)诱导的气管上皮细胞向间质细胞转化(EMT)中的表达情况,探讨miR-152能否影响EMT的发生。方法:用10ng/mL的TGF-β1诱导人气管上皮16HBE细胞EMT发生,光镜下观察细胞形态变化,qRT-PCR和Westernblot检测细胞中EMT相关标志物α平滑肌肌动蛋白(α-SMA)、E-cadherin和Vimentin的改变;qRT-PCR检测有或无TGF-β1处理的16HBE细胞中miR-152的表达情况;通过转染miR-152mimics和miR-NC至细胞后,再次检测细胞中EMT标志物的表达情况。结果:与对照组比,TGF-β1处理72h时能引起16HBE细胞发生EMT,细胞形态在72h改变明显,α-SMA和Vimentin表达增加,E-cadherin表达下降(P<0.01);转染miR-152mimics,细胞内miR-152的表达显著增加(P<0.01);与miR-NC组比,转染miR-152过表达后,气管上皮细胞中α-SMA和Vimentin的表达降低(P<0.01),E-cadherin表达增加(P<0.01),EMT受到抑制。结论:TGF-β1能诱导离体培养的气管上皮细胞发生EMT;miR-152可以抑制TGF-β1诱导的气管上皮细胞EMT的发生。

Objective： To verify whether transforming growth factor-beta1（TGF-β1） could induce epithelial-mesenchymal transition（EMT） in vitro and to determine the role of miR-152 in TGF-β1-mediated EMT in bronchial epithelial cells. Methods： 10 ng/mL of TGF-β1 was used to induce the EMT of human bronchial epithelial cells 16 HBE. Morphological changes were observed and q RT-PCR and Western blot were employed to test the alteration of αSMA, E-cadherin and Vimentin, thus to verify the occurrence of EMT. q RT-PCR was then used to test the expression profile of miR-152 in TGF-β1 treated 16 HBE cells. After miR-152 was successfully transfected into 16 HBE cells, EMT related markers were then detected using q RT-PCR and Western blot. Results： Compared with control group, TGF-β1-treated cells presented a myofibroblast-like morphology, especially at 72 h, characteristic by the loss of cell-to-cell junction, denovo expression of α-SMA and Vimentin, and loss of epithelial marker E-cadherin（P0.01）. The expression levels of miR-152 in TGF-β1 treated 16 HBE cells were dramatically increased after transfected with miR-152 mimics（P0.01）. Compared with the control group, α-SMA and Vimentin were decreased, whereas E-cadherin was increased in the miR-152 group（P0.01）, suggesting overexpression of miR-152 could reverse the EMT induced by TGF-β1 in 16 HBE cells. Conclusion：TGF-β1 induces EMT of human bronchial epithelial cells in vitro; miR-152 inhibits TGF-β1-mediated EMT in 16 HBE cells.