下调miR-215-5p对TGF-β诱导的人肺成纤维细胞IMR-90增殖抑制作用观察  

Inhibitory effect of down-regulation of miR-215-5p on TGF-β-induced proliferation of human lung fibroblasts IMR-90

作　　者：杜伟 郭丽娜 庞席宁 DU Wei;GUO Lina;PANG Xining(Liaoning Zhongzhi Shengjing Geriatric Hospital,Shenyang 110000,China)

机构地区：[1]辽宁中置盛京老年病医院,沈阳110000 [2]中国医科大学附属盛京医院 [3]沈阳军区总医院浑南分院

出　　处：《山东医药》2018年第40期15-19,共5页Shandong Medical Journal

摘　　要：目的探讨下调miR-215-5p对TGF-β诱导的人肺成纤维细胞IMR-90增殖的影响及其机制。方法将体外培养的IMR-90细胞分为对照组、TGF-β组、TGF-β+NC inhibitor组、TGF-β+miR-215-5p inhibitor组。对照组细胞正常培养; TGF-β组给予10 ng/m L的TGF-β处理细胞; TGF-β+NC inhibitor组和TGF-β+miR-215-5p inhibitor组细胞先采用NC inhibitor或者miR-215-5p inhibitor转染细胞,24 h后加入10 ng/m L的TGF-β处理细胞。采用MTT法检测各组在转染后24、48、72、96 h的细胞增殖能力;流式细胞术检测转染48 h后细胞周期变化情况;荧光定量PCR和Western blotting法检测转染48 h后各组细胞cyclin D1、RB1和α-SMA的表达。结果转染后48、72、96 h,TGF-β组细胞增殖能力高于对照组(P均<0. 05),TGF-β+miR-215-5p inhibitor组细胞增殖能力低于TGF-β+NC inhibitor组(P均<0. 05)。与对照组相比,TGF-β组G1期细胞比例降低,S期细胞比例升高(P均<0. 05);与TGF-β+NC inhibitor组相比,TGF-β+miR-215-5p inhibitor组G1期细胞比例增加,S期细胞比例降低(P均<0. 05)。与对照组相比,TGF-β组cyclin D1、α-SMA的表达升高,RB1的表达降低(P均<0. 05);与TGF-β+NC inhibitor组相比,TGF-β+miR-215-5p inhibitor组cyclin D1、α-SMA的表达降低,RB1的表达升高(P均<0. 05)。结论下调miR-215-5p对TGF-β诱导的人肺成纤维细胞IMR-90增殖具有抑制作用。该作用可能原因为下调miR-215-5p能够抑制RB1表达进而下调cyclin D1,阻碍TGF-β诱导的肺成纤维细胞的周期转换进而抑制肺成纤维细胞的增殖;同时,下调miR-215-5p也能够抑制α-SMA的表达阻碍TGF-β诱导的肺成纤维细胞向肌成纤维细胞转化。Objective To investigate the inhibitory effect of down-regulation of miR-215-5p on TGF-β-induced proliferation of human lung fibroblasts IMR-90 and its mechanism.Methods Lung fibroblasts MIR-90 were divided into four groups:the control group,TGF-βgroup,TGF-β+NC inhibitor group,and TGF-β+miR-215-5p inhibitor group.Cells in the TGF-βgroup were treated with 10 ng/ml TGF-β,cells in the TGF-β+NC inhibitor group or TGF-β+miR-215-5p inhibitor group were transfected with NC inhibitor or miR-215-5p inhibitor,then treated with 10 ng/ml TGF-β.After that,the cell proliferation at 24,48,72 and 96 h was detected by MTT assay,the cell cycle was determined by flow cytometry,and the expression of cyclinD1,retinoblastoma gene 1(RB1),andα-smooth muscle actin(α-SMA)was examined by real-time PCR and Western blotting.Results At 48,72,and 96 h after transfection,the cell proliferation ability of the TGF-βgroup was higher than that of the control group(P<0.05),and the cell proliferation ability of TGF-β+miR-215-5p inhibitor group was lower than that of TGF-β+NC inhibitor group(P<0.05).Compared with the control group,the proportion of cells in the G 1 phase of the TGF-βgroup decreased,and the proportion of cells in the S phase increased(both P<0.05).Compared with the TGF-β+NC inhibitor group,the proportion of cells in the G 1 phase of the TGF-β+miR-215-5p inhibitor group increased,and the proportion of cells in the S phase decreased(both P<0.05).Compared with the control group,the expression of cyclinD1 andα-SMA in the TGF-βgroup increased,and the expression of RB1 decreased(all P<0.05).Compared with TGF-β+NC inhibitor group,the expression of cyclinD1 andα-SMA decreased and the expression of RB1 increased in the TGF-β+miR-215-5p inhibitor group(all P<0.05).Conclusions Down-regulation of miR-215-5p has an inhibitory effect on TGF-β-induced proliferation of human lung fibroblasts IMR-90.The mechanism may be that the down-regulation of miR-215-5p inhibits the RB1 expression and thus down-regulates cyclinD1,blocks TGF

关 键 词：miRNA-215-5p 肺纤维化 细胞增殖 细胞周期蛋白 视网膜母细胞瘤基因1 Α-平滑肌肌动蛋白 

分 类 号：R563[医药卫生—呼吸系统]