miR-21对TGF-β1诱导的肾小管上皮细胞间质转分化的影响

doi:10.12047/j.cjap.5477.2017.084

中国应用生理学杂志 ›› 2017, Vol. 33 ›› Issue (4): 346-350.doi: 10.12047/j.cjap.5477.2017.084

miR-21对TGF-β1诱导的肾小管上皮细胞间质转分化的影响

蔡月琴, 褚燕青, 朱科燕, 陈诚, 王德军

浙江中医药大学动物实验研究中心/比较医学研究所, 杭州 315700

收稿日期:2016-07-21 修回日期:2017-01-30 出版日期:2017-07-28 发布日期:2018-06-19
通讯作者: 王德军,Tel:0571-86613662;E-mail:wdj0369@126.com E-mail:wdj0369@126.com
基金资助:
浙江省自然科学基金资助项目（LQ14H290004）；浙江中医药大学比较医学创新团队资助项目（XTD201301）

The influence of miR-21 on HK-2 EMT cells induced by TGF-beta 1

CAI Yue-qin, CHU Yan-qing, ZHU Ke-yan, CHEN Cheng, WANG De-jun

Zhejiang Chinese Medical University, Laboratory Animal Research Center/Comparative Medicine Institude, Hangzhou 310053, China

Received:2016-07-21 Revised:2017-01-30 Online:2017-07-28 Published:2018-06-19
Supported by:
浙江省自然科学基金资助项目（LQ14H290004）；浙江中医药大学比较医学创新团队资助项目（XTD201301）

摘要/Abstract

摘要： 目的：观察miR-21在转化生长因子β1（TGF-β1）诱导的人肾小管上皮细胞（HK-2细胞）上皮间质转分化（EMT）中的作用，并探讨miR-21参与调控HK-2细胞EMT的可能靶点。方法：体外培养的HK-2细胞分为6组：正常对照组、转化生长因子β1（TGF-β1）模型组、miR-21 mimic阴性组、miR-21 mimic组、miR-21 inhibitor阴性组和miR-21 inhibitor组。细胞经4 ng/ml TGF-β1处理建立EMT模型，检测miR-21和EMT相关指标的表达变化，利用基因转染技术，将miR-21 mimic质粒或miR-21 inhibitor质粒转染经TGF-β1处理的HK-2细胞，使细胞过表达或抑制表达miR-21，在此基础上观察细胞EMT相关指标的变化以及磷酸酯酶（PTEN）基因的影响。结果：①与正常组相比，模型组的miR-21含量显著升高（P<0.05），上皮表型标志物E-cadherin的mRNA和蛋白表达水平均显著降低（P<0.01），间质表型标志物α平滑肌肌动蛋白（α-SMA） mRNA和蛋白水平也显著升高（P<0.05，P<0.01）；②转染miR-21 mimic后，与miR-21 mimic阴性对照组相比，miR-21含量显著升高（P<0.01），PTEN、E-cadherin的mRNA和蛋白水平显著降低（P<0.05，P<0.01），α-SMA mRNA和蛋白水平显著升高（P<0.05，P<0.01）；转染miR-21 inhibitor后，与miR-21 inhibitor阴性对照组相比，miR-21含量显著降低（P<0.01），PTEN、E-cadherin的mRNA和蛋白含量显著升高（P<0.05，P<0.01），α-SMA mRNA、蛋白水平显著降低（P<0.05，P<0.01）。结论：miR-21在TGF-β1诱导的HK-2细胞EMT发生中具有重要作用，并且可能通过靶基因PTEN参与EMT相关分子的表达调控。

关键词: 微小RNA-21, HK-2细胞, 转化生长因子-β1, 上皮间质转分化

Abstract: Objective: To investigate the effect of the miR-21 and its target mRNA in renal tubular epithelial mesenchymal transformation (EMT) model induced by transformation growth factor-β1(TGF-β1) in human renal tubular epithelial (HK-2) cells.Methods: HK-2 cells were divided into 6 groups:normal control group, TGF-β1 group, miR-21 mimic negative group, miR-21 mimic group, miR-21 inhibitor negative group and miR-21 inhibitor group. EMT model was established in HK-2 cells induced by 4 ng/ml TGF-β1. The level of miR-21, the mRNA and protein expression of EMT related factors were detected. MiR-21 mimic plasmid and miR-21 inhibitor plasmid were transfected into HK-2 cells that treated with TGF-β1 respectively using liposome transfection technique. Observe the impact of overexpression or inhibition expression of miR-21 on the mRNA and protein expression of EMT related factors and PTEN.Results: ①Compared with the normal group, the level of miR-21 was significantly increased in model group (P<0.05), the mRNA and protein expression levels of epithelial cells marker E-cadherin was significantly decreased (P<0.01), while the mRNA and protein levels of mesenchymal cells marker α-SMA was significantly increased (P<0.05,P<0.01). ②Compared with the miR-21 mimic negative group, the level of miR-21 in miR-21 mimic group increased significantly (P<0.01), the mRNA and protein expression levels of PTEN and E-cadherin decreased significantly (P<0.05,P<0.01), the mRNA and protein levels of α-SMA increased significantly (P<0.05,P<0.01). Compared with the miR-21 inhibitor negative control group, the level of miR-21 in miR-21 inhibitor group decreased significantly (P<0.01), the mRNA and protein expression levels of PTEN and E-cadherin increased significantly (P<0.05,P<0.01), the mRNA and protein levels of α-SMA decreased significantly (P<0.05,P<0.01).Conclusion: MiR-21 may play an important role in EMT induced by TGF-β1 in HK-2 cells and regulate the expression of EMT related factors its target gene PTEN.

Key words: microRNA-21, HK-2 cells, transformation growth factor-β1, epithelial mesenchymal transformation

蔡月琴, 褚燕青, 朱科燕, 陈诚, 王德军. miR-21对TGF-β1诱导的肾小管上皮细胞间质转分化的影响[J]. 中国应用生理学杂志, 2017, 33(4): 346-350.

CAI Yue-qin, CHU Yan-qing, ZHU Ke-yan, CHEN Cheng, WANG De-jun. The influence of miR-21 on HK-2 EMT cells induced by TGF-beta 1[J]. CJAP, 2017, 33(4): 346-350.

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miR-21对TGF-β1诱导的肾小管上皮细胞间质转分化的影响

The influence of miR-21 on HK-2 EMT cells induced by TGF-beta 1

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