miR-200a抑制YAP1基因表达对肺癌细胞增殖的影响

发布时间：2018-04-18 18:37

本文选题：miR-a + 肺癌　；参考：《中国肿瘤临床》2017年07期

【摘要】：目的:研究微小RNA-200a(miR-200a)对肺癌细胞增殖的影响,并探讨其分子机制。方法:采用Real-time PCR检测15例非小细胞肺癌组织和对应癌旁组织、人肺癌细胞株(A549、NCI-H520、SK-MES-1)及人正常肺支气管上皮细胞株16HBE中miR-200a的表达水平。用CCK-8法检测miR-200a对A549肺癌细胞增殖活性的影响。通过生物信息学方法预测miR-200a可能的靶基因,双荧光素酶报告基因实验结合Real-time PCR和Western blot验证miR-200a对靶基因YAP1的调控作用。CCK-8法检测下调靶基因YAP1对A549肺癌细胞株增殖活性的影响。结果:miR-200a在非小细胞肺癌组织和肺癌细胞系中表达明显降低(P0.01)。上调miR-200a表达后明显抑制A549肺癌细胞的增殖活力(P0.01)。双荧光素酶报告基因显示miR-200a可以直接作用于靶基因YAP1的3'-UTR区域抑制荧光素酶活性(P0.01),Real-time PCR和Western blot检测显示上调miR-200a的表达能够明显下调A549肺癌细胞YAP1 mRNA和蛋白的表达水平(P0.01)。CCK-8法显示下调YAP1的表达能够明显抑制A549肺癌细胞的增殖活性(P0.01)。结论:miR-200a通过靶向作用于YAP1基因来抑制肺癌细胞的增殖,从而在肺癌中发挥抑癌基因的功能。
[Abstract]:Aim: to study the effects of microRNA-200a- miR-200a on the proliferation of lung cancer cells and its molecular mechanism.Methods: Real-time PCR was used to detect the expression of miR-200a in 15 cases of non-small cell lung cancer (NSCLC) and its adjacent tissues, human lung cancer cell line A549 (NCI-H520SK-MES-1) and human normal pulmonary bronchial epithelial cell line (16HBE).The effect of miR-200a on proliferation of A549 lung cancer cells was detected by CCK-8 assay.The possible target genes of miR-200a were predicted by bioinformatics. The double luciferase reporter gene experiment combined with Real-time PCR and Western blot was used to verify the effect of miR-200a on the target gene YAP1. CCK-8 method was used to detect the effect of down-regulating target gene YAP1 on the proliferation of A549 lung cancer cell line.Results the expression of 1% miR-200a was significantly decreased in non-small cell lung cancer tissues and lung cancer cell lines.After upregulation of miR-200a expression, the proliferative activity of A549 lung cancer cells was significantly inhibited (P 0.01).Double luciferase reporter gene showed that miR-200a could directly inhibit luciferase activity in 3'-UTR region of target gene YAP1 Real-time PCR and Western blot showed that up-regulation of miR-200a expression could significantly down-regulate the expression of YAP1 mRNA and protein in A549 lung cancer cell line.The down-regulated expression of YAP1 could significantly inhibit the proliferation activity of A549 lung cancer cells.ConclusionTwo miR-200a can inhibit the proliferation of lung cancer cells by targeting YAP1 gene and thus play the role of tumor suppressor gene in lung cancer.
【作者单位】：新汶矿业集团中心医院肿瘤科;山东大学附属济南市中心医院胸外科;
【分类号】：R734.2

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