[微小RNA-21对转化生长因子-β1诱导的人肾小管上皮细胞系HK-2上皮-间质转化的影响]
[The influence of miR-21 on HK-2 EMT cells induced by TGF-beta 1].
作者信息
Cai Yue-Qin, Chu Yan-Qing, Zhu Ke-Yan, Chen Cheng, Wang De-Jun
机构信息
Zhejiang Chinese Medical University, Laboratory Animal Research Center/Comparative Medicine Institude, Hangzhou 310053, China.
出版信息
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2017 Apr 8;33(4):346-350.
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 (<0.05), the mRNA and protein expression levels of epithelial cells marker E-cadherin was significantly decreased (<0.01), while the mRNA and protein levels of mesenchymal cells marker α-SMA was significantly increased (<0.05,<0.01). ②Compared with the miR-21 mimic negative group, the level of miR-21 in miR-21 mimic group increased significantly (<0.01), the mRNA and protein expression levels of PTEN and E-cadherin decreased significantly (<0.05,<0.01), the mRNA and protein levels of α-SMA increased significantly (<0.05,<0.01). Compared with the miR-21 inhibitor negative control group, the level of miR-21 in miR-21 inhibitor group decreased significantly (<0.01), the mRNA and protein expression levels of PTEN and E-cadherin increased significantly (<0.05,<0.01), the mRNA and protein levels of α-SMA decreased significantly (<0.05,<0.01).
CONCLUSIONS
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.
目的
探讨微小RNA-21（miR-21）及其靶mRNA在转化生长因子-β1（TGF-β1）诱导的人肾小管上皮（HK-2）细胞肾小管上皮细胞间质转化（EMT）模型中的作用。
方法
将HK-2细胞分为6组：正常对照组、TGF-β1组、miR-21模拟物阴性组、miR-21模拟物组、miR-21抑制剂阴性组和miR-21抑制剂组。用4 ng/ml TGF-β1诱导HK-2细胞建立EMT模型。检测miR-21水平、EMT相关因子的mRNA和蛋白表达。采用脂质体转染技术分别将miR-21模拟物质粒和miR-21抑制剂质粒转染至经TGF-β1处理的HK-2细胞。观察miR-21过表达或抑制表达对EMT相关因子和PTEN的mRNA及蛋白表达的影响。
结果
①与正常组比较，模型组miR-21水平显著升高（<0.05），上皮细胞标志物E-钙黏蛋白的mRNA和蛋白表达水平显著降低（<0.01），而间充质细胞标志物α-平滑肌肌动蛋白（α-SMA）的mRNA和蛋白水平显著升高（<0.05，<0.01）。②与miR-21模拟物阴性组比较，miR-21模拟物组miR-21水平显著升高（<0.01），PTEN和E-钙黏蛋白的mRNA及蛋白表达水平显著降低（<0.05，<0.01），α-SMA的mRNA和蛋白水平显著升高（<0.05，<0.01）。与miR-21抑制剂阴性对照组比较，miR-21抑制剂组miR-21水平显著降低（<0.01），PTEN和E-钙黏蛋白的mRNA及蛋白表达水平显著升高（<0.05，<0.01），α-SMA的mRNA和蛋白水平显著降低（<0.05，<0.01）。
结论
miR-21可能在TGF-β1诱导的HK-2细胞EMT中起重要作用，并调节EMT相关因子及其靶基因PTEN的表达。
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