miR-200c阻滞转化生长因子-β1在腹膜后成纤维细胞增殖和迁移侵袭中的作用  

Effect of miR-200c retardation transforming growth factor-β1 in retroperitoneal fibroblast proliferation and migration

作　　者：施莹[1] 林玲[1] 阚佳音 刘海燕[2] 马增伟[1] 李双宇[1] 

机构地区：[1]齐齐哈尔医学院第三附属医院肾内科,齐齐哈尔161000 [2]齐齐哈尔医学院病理学院微形态实验中心,齐齐哈尔161000

出　　处：《中国免疫学杂志》2017年第7期1005-1008,共4页Chinese Journal of Immunology

基　　金：齐齐哈尔市科学技术计划项目任务(FSGG201422)

摘　　要：目的:阐明miR-200c对腹膜后成纤维细胞增殖的影响,并分析其分子机制,为抑制腹膜后纤维化提供理论依据。方法:收集36例人腹膜后组织,原代培养成纤维细胞,以5 ng/ml TGF-β1刺激者为TGF-β1组,pc DNA3.1-miR-200c转染成纤维细胞,再给予5 ng/ml TGF-β1刺激者为miR-200c组,仅以pc DNA3.1质粒转染者为pc DNA3.1质粒组,正常培养的成纤维细胞为对照组。分别以噻唑蓝(MTT)细胞增殖实验、细胞划痕实验、Transwell细胞小室迁移实验检测成纤维细胞增殖和迁移能力,并以ELISA实验检测各组细胞裂解液中Akt蛋白的含量。结果:miR-200c组成纤维细胞的增殖OD值明显比TGF-β1组降低(P<0.01);与TGF-β1组相比,miR-200c组细胞迁移率明显降低(P<0.01);miR-200c组Akt蛋白的含量比TGF-β1组明显降低(P<0.01)。结论:miR-200c下调Akt通路而抑制TGF-β1对成纤维细胞的增殖或迁移效果,对抑制腹膜纤维化具有重要意义。Objective： To clarify the effect of miR-200c on the proliferation of retroperitoneal fibroblast, and analyze its molecular mechanism, which provide theoretical basis for the inhibition of retroperitoneal fibrosis. Methods： 36 cases of retroperitoneal tissues were collected, and the primary fibroblasts were cultured. The cells stimulated by 5 ng/ml transforming growth factor-β1 （TGF- β1 ） were used as TGF-β1 group;the cells were transfected with pcDNA3. 1-miR-200c first, and then stimulated by 5 ng/ml TGF-β1 were used as miR-200c group;the cells transfected only by peDNA 3.1 were used as pcDNA 3. 1 group;the normal cultured cells were used as control group. The The assay, cell scratch and Transwell chamber were used to detect the proliferation and migration of fibroblast, respectively. The ELISA experiment was used to detect the content of Akt protein in the lysis liquid of each group. Results： The OD value of fibroblast proliferation in miR-200e group was obviously lower than that of TGF-β1 group （P〈0.01） ;compared with TGF-β1 group, the cell migration rate was decreased significantly in miR-200c group （P〈0.01） ;the content of Akt protein in miR- 200c was obviously lower than that of TGF-β1 group （P〈0.01）. Conclusion： miR-200c inhibits the effect of TGF-β1 on the proliferation and migration of fibroblast by down-regulating Akt signaling pathway, which has important significance in inhibiting peritoneal fibrosis.

关 键 词：MIR-200C 成纤维细胞 增殖 迁移 

分 类 号：R692.9[医药卫生—泌尿科学]