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

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摘要目的：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10 ng/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Western blot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-152 mimics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72 h时能引起16HBE细胞发生EMT，细胞形态在72 h改变明显，α-SMA和Vimentin表达增加，E-cadherin表达下降（P＜0.01）；转染miR-152 mimics，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。

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	朱融和
	孙媛媛
	钱燕

关键词 ：哮喘, 气管上皮细胞, 间质细胞, miR-152

Abstract：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 16HBE. Morphological changes were observed and qRT-PCR and Western blot were employed to test the alteration of αSMA, E-cadherin and Vimentin, thus to verify the occurrence of EMT. qRT-PCR was then used to test the expression profile of miR-152 in TGF-β1 treated 16HBE cells. After miR-152 was successfully transfected into 16HBE cells, EMT related markers were then detected using qRT-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 (P<0.01). The expression levels of miR-152 in TGF-β1 treated 16HBE cells were dramatically increased after transfected with miR-152 mimics (P<0.01). Compared with the control group, α-SMA and Vimentin were decreased, whereas E-cadherin was increased in the miR-152 group (P<0.01), suggesting overexpression of miR-152 could reverse the EMT induced by TGF-β1 in 16HBE cells. Conclusion: TGF-β1 induces EMT of human bronchial epithelial cells in vitro; miR-152 inhibits TGF-β1-mediated EMT in 16HBE cells.

Key words： asthma bronchial epithelial cells mesenchymal cells miR-152

收稿日期: 2017-04-13

基金资助:温州市科技计划项目（2014Y0143）。

通讯作者: 钱燕，主任医师，硕士生导师，Email：qianyan_paw@163.com

作者简介: 朱融和（1985-），女，浙江温州人，住院医师，硕士

链接本文:

https://xb.wmu.edu.cn/CN/ 或 https://xb.wmu.edu.cn/CN/Y2017/V47/I10/739

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