LncRNA SIL对肺泡上皮细胞增殖和迁移的抑制作用研究

Inhibitory effect of lncRNA SIL on proliferation and migration of alveolar epithelial cells

目的研究lncRNA SIL与肺纤维化过程中肺泡上皮细胞的增殖和迁移之间的关系。方法利用RNA fish研究在转化生长因子(TGF-β1)诱导肺泡上皮细胞向间质细胞转化的过程中lncRNA SIL的表达变化,构建lncRNA SIL真核表达载体,转染A549细胞后,通过RT-PCR、MTT、Western blot、细胞损伤修复以及免疫荧光实验检测过表达lncRNA SIL对细胞A549增殖和迁移的作用以及对相关标志蛋白表达的影响。结果当使用TGF-β1诱导48 h后,细胞中lncRNA SIL的表达量与对照组相比降低;在A549细胞中过表达lncRNA SIL时,细胞增殖能力、迁移能力以及细胞增殖相关蛋白PCNA和Ki67的表达与对照组相比均降低;对细胞标志蛋白的表达分析结果显示,与对照组比,转染lncRNA SIL组,间质细胞标志蛋白α-SMA和Collagen-1表达降低,肺泡上皮细胞标志蛋白E.cad表达升高。结论LncRNA SIL可以通过抑制A549细胞的增殖和迁移达到抑制肺泡上皮细胞向间质细胞转化的作用。

Objective To study the relationship between lncRNA SIL and proliferation and migration of alveolar epithelial cell during pulmonary fibrosis.Methods The expression level of lncRNA SIL in A549 cells induced by transforming growth factor(TGF-β1)was detected by RNA fish.The eukaryotic expression vector of lncRNA SIL was built and transfected into A549 cells,the proliferation and migration of the cells after overexpressing lncRNA SIL were studied by RT-PCR,MTT,Western blot,cell damage repair experiments and immunofluorescence.Results The migration and proliferation of A549 cells were significantly reduced compared with the control group after transfected by lncRNA SIL.When lncRNA SIL was overexpressed in A549 cells,cell proliferation,migration and expression of proliferation-related proteins PCNA and Ki67 were decreased compared with the control group.Analysis of the expression of cellular marker proteins showed that,the expression levels of interstitial cell marker proteinsα-SMA and Collagen-1 were significantly reduced,while the expression level of alveolar epithelial marker protein E.cad significantly increased.Conclusion LncRNA SIL can inhibit the proliferation and migration of A549 cells,and also can inhibit EMT induced by TGF-β1.