miR-302c靶向CREB1基因通过P53信号通路影响肺癌的侵袭和迁移

miR-302c affects invasion and migration of lung cancer by targeting CREB1 gene via P53 signaling pathway

目的:探讨微小RNA-302c(miR-302c)在肺癌组织中的表达,以及对肺癌侵袭和迁移的影响和作用机制。方法:在线分析GEO数据库中GSE19945和GSE136043两组肺癌数据集中miR-302c的表达情况,通过Human Protein Atlas数据库研究CREB1表达情况;双荧光素酶实验证明miR-302c和CREB1的关系。正常组细胞不加任何药物,对照组细胞转染miR-302cmimic-NC,实验组细胞转染miR-302c-mimic。通过Transwell小室检测细胞的侵袭与迁移能力,通过小管形成检测细胞的血管生成能力,通过Western blot检测各组细胞中CREB1、p-P53、p-P21的表达水平。结果:生物信息分析显示,与正常组织相比,miR-302c在肺癌组织、肺癌淋巴转移组织中的表达明显降低,肺癌组织中CREB1的表达明显升高;双荧光素酶实验证明miR-302c靶向调控CREB1的表达;与正常组相比,实验组迁移和侵袭的细胞数量、小管生成的数量明显下降,实验组中CREB1的表达明显下降,p-P53、p-P21的表达明显升高(均P<0.05)。结论:miR-302c在肺癌组织表达降低,过表达miR-302c后能明显抑制肺癌细胞的侵袭与转移,这可能与抑制靶基因CREB1的表达,以及激活P53信号通路有关。

Objective:To investigate the expression of microRNA-302c(miR-302c)in lung cancer,assess its influence on the invasion and migration of lung cancer,and examine the underlying mechanism.Methods:miR-302c expression in two lung cancer datasets(GSE19945 and GSE136043)retrieved from the online database GEO was statistically analyzed,and CREB1 expression was screened using the Human Protein Atlas.A dual luciferase assay was used to assess interactions between miR-302c and CREB1.Normal cells were cultured without any drugs;these controls were transfected with miR-302c-mimic-NC.Experimental cells were transfected with miR-302c-mimic.A transwell chamber assay was used to assess the invasion and migration ability;a tubule formation experiment was used to detect the angiogenesis ability.Western blot was used to measure the expression levels of CREB1,p-P53,and p-P21.Results:Online bioinformatics analyses showed that,compared with normal tissues,miR-302c expression was significantly reduced in lung cancer tissue and lung cancer lymphatic metastasis tissue,and CREB1 expression in lung cancer tissue was significantly increased.Dual luciferase assays showed that miR-302c regulated CREB1 expression;compared with the controls,the number of cells that migrated and invaded and the number of tubules were significantly decreased in the treatment group,CREB1 was significantly decreased in treated cells,and expression of p-P53 and p-P21 was increased(P<0.05,each).Conclusions:miR-302c expression in lung cancer tissue is reduced.Overexpression of miR-302c can significantly inhibit the invasion and metastasis of lung cancer cells,which may be related to reduced CREB1 expression and activation of the P53 signaling pathway.