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

发布时间：2019-06-11 21:02

【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(P0.01);与TGF-β1组相比,miR-200c组细胞迁移率明显降低(P0.01);miR-200c组Akt蛋白的含量比TGF-β1组明显降低(P0.01)。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。
[Abstract]:Aim: to elucidate the effect of miR-200c on the proliferation of retroperitoneal fibroblasts and analyze its molecular mechanism, so as to provide theoretical basis for inhibiting retroperitoneal fibrosis. Methods: 36 cases of human retroperitoneal tissue were collected. The primary cultured fibroblasts were stimulated with 5 ng/ml TGF- 尾 1 as TGF- 尾 1 group, pc DNA3.1-miR-200c was transformed into fibroblasts, miR-200c group was stimulated with 5 ng/ml TGF- 尾 1, pc DNA3.1 plasmid was only infected with pc DNA3.1 plasmid, and normal cultured fibroblasts were used as control group. The proliferation and migration ability of fibroblasts were detected by thiazolyl blue (MTT) cell proliferation test, cell scratch test and Transwell cell chamber migration test, and the content of Akt protein in cell lysate of each group was detected by ELISA assay. Results: the proliferation of fibroblasts in miR-200c group was significantly lower than that in TGF- 尾 1 group (P 0.01), the cell mobility in miR-200c group was significantly lower than that in TGF- 尾 1 group (P 0.01), and the content of Akt protein in miR-200c group was significantly lower than that in TGF- 尾 1 group (P 0.01). Conclusion: miR-200c down-regulate Akt pathway and inhibit the proliferation or migration of fibroblasts by TGF- 尾 1, which is of great significance in inhibiting peritoneal fibrosis.
【作者单位】：齐齐哈尔医学院第三附属医院肾内科;齐齐哈尔医学院病理学院微形态实验中心;
【基金】：齐齐哈尔市科学技术计划项目任务(FSGG201422)
【分类号】：R57

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