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

作　　者：蔡月琴[1] 褚燕青 朱科燕[1] 陈诚[1] 王德军[1] 

机构地区：[1]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

出　　处：《中国应用生理学杂志》2017年第4期346-350,共5页Chinese Journal of Applied Physiology

基　　金：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：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。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-j31 （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 nega- tive group and miR-21 inhibitor group. EMT model was established in HK-2 cells induced by 4 ng/ml TGF-131. The level of miR-21, the mR- NA 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 transfeetion technique. Observe the impact of overexpression or itdaibition ex- pression of miR-21 on the mlLNA and protein expression of EMT related factors and PTEN. Results： （2）Compared 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 mesenchyrnal cells marker a-SMA was significantly in- creased （P 〈 0.05, P 〈 0.01）. （2）Compared with the miR-21 minaic negative group, the level of miR-21 in miR-21 mimic group increased significantly （ P 〈 0.01 ）, the rnRNA and protein expression levels of PTEN and E-cadherin decreased significantly （ P 〈 0.05, P 〈 0.01 ）, the mRNA and protein levels of a-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 mR_NA and protein expression levels of PTEN and E-cadherin increased significantly （ P 〈 0.05, P 〈 0.01）, the mRNA and protein levels of a-SMA decreased significantly （ P 〈 0.05, P 〈 0.01 ）. Conclusion： MiR-21 may play

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 

分 类 号：R392.11[医药卫生—免疫学]