CSE诱导支气管上皮细胞分泌外泌体miR-186促进肺成纤维细胞增殖

Cigarette smoke extract-induced 16HBE-derived exosomal miR-186 significantly promoted the proliferation of COPD MRC-5 cell

目的:研究香烟提取物(cigarette smoke extract,CSE)对支气管上皮细胞16HBE外泌体miR-186表达量的影响,以及16HBE来源的外泌体miR-186对肺成纤维细胞增殖的影响。方法:收集CSE处理的16HBE细胞上清中外泌体miR-186,用于MRC-5细胞培养;使用qPCR检测外泌体miR-186的表达;使用CCK-8检测MRC-5细胞增殖能力;使用双荧光素酶验证miR-186与Bcl2L11基因的靶向结合,使用Western blot检测Bcl2L11的表达。结果:透射电镜观察到16HBE细胞上清外泌体直径约100 nm的球形结构形态;CSE处理明显促进外泌体miR-186的表达;CSE诱导的外泌体miR-186能促进MRC-5细胞增殖;miR-186与Bcl2L11存在靶向结合关系;miR-186 mimic明显抑制了Bcl2L11的表达。结论:CSE诱导的16HBE细胞外泌体miR-186通过靶向结合Bcl2L11基因,在成纤维细胞增殖中起到重要作用。

Objective:To study the effect of cigarette smoke extract(CSE)on the expression of exosomal miR-186 derived from 16 HBE cells,and the effects of 16 HBE-derived exosomal miR-186 on the proliferation of MRC-5 cells.Methods:This study collected the exosomal miR-186 in the supernatant of CSE-treated 16 HBE cells for MRC-5 cell culture;the expression of exosomal miR-186 was detected by qPCR;the proliferation of MRC-5 cell was detected by CCK-8;dual luciferase reporter gene experiment was used to detect the targeted regulation relationship between miR-186 and Bcl2 L11;the expression of Bcl2 L11 was detected by Western blot.Results:The morphology of 16 HBE cells about 100 nm in diameter was observed by TEM;CSE treatment significantly promoted the expression of exosomal miR-186;CSE-induced exosomal miR-186 promoted the proliferation of MRC-5 cells;Bcl2 L11 was a target gene of miR-186;miR-186 mimics significantly decreased Bcl2 L11 expression.Conclusion:CSE-induced 16 HBE-derived exosomal miR-186 promotes the proliferation of MRC-5 cells by target binding Bcl2 L11 genes.