摘要
目的分析miRNA-200a对肺癌细胞增殖影响及抑癌基因功能影响。方法 Real-time PCR对正常肺支气管上皮细胞16HBE、肺癌细胞SK-MES-1、NCI-H520、A549及25例非小细胞癌癌旁组织与癌组织内miR-200a表达量检测,CCK-8法检测肺癌A549细胞增殖活性受miRNA-200a影响状况,生物信息学对miRNA-200a靶基因进行预测,双荧光素酶联合Western blot及Real-time PCR检验YAP1受miRNA-200a调控影响。CCK-8法检测肺癌549细胞受YAP1增殖影响。结果肺癌细胞株SK-MES-1、NCI-H520及A549内miR-200a表达量均低于正常细胞株16HBE,差异有统计学意义(P<0.05)。miR-200a mimics组内A549细胞在48、72及96 h时其吸光度均低于Mimics-NC,差异有统计学意义(P<0.05)。肺癌A549转染siRNA-NC或者siRNA-YAP1后,PCR检测显示siRNA-YAP1内YAP1 m RNA表达量为(0.37±0.06),siRNA-NC内YAP1 mRNA表达量为(1.03±0.07),差异有统计学意义(P<0.05);Western blot显示siRNA-YAP1内YAP1蛋白表达量比siRNA-NC低;siRNA-YAP1内YAP1细胞吸光度在48、72及96 h时均低于siRNA-NC,差异有统计学意义(P<0.05)。结论 miR-200a对肺癌细胞增殖的抑制作用主要是经过靶向作用YAP1基因来实现的,进而在肺癌内起到抑癌功能。
Objective To analyze the effects of miRNA-200a on the proliferation of lung cancer cells and the function of tumor suppressor genes. Methods Real-time PCR was used to detect the expression quantity of miR-200a in normal lung bronchial epithelial cells 16HBE, lung cancer cells SK-MES-1, NCI-H520, A549 and non-small cell carcinoma para-carcinoma tissue tissues and cancer tissues in 25 cases. CCK-8 assay was used to detect the effect of miRNA-200a on the proliferation activity of lung cancer A549 cells. Bioinformatics was used to predict miRNA-200a target genes, and dual fluorescein combined with Western blot and Real-time PCR were used to detect the effect of miRNA-200a regulation on YAP1. CCK-8 assay was used to detect the effect of YAP1 proliferation on lung cancer 549 cells. Results The expression quantity of miR-200a in lung cancer cell lines SK-MES-1, NCI-H520 and A549 was lower than that in normal cell line 16HBE, and the difference was statistically significant（P〈0.05）. The absorbance of A549 cells in miR-200a mimics group was lower than that of Mimics-NC at 48, 72 and 96 h, and the difference was statistically significant（P〈0.05）. After lung cancer A549 was transfected with siRNA-NC or siRNA-YAP1, PCR detection showed that the expression quantity of YAP1 mRNA in siRNA-YAP1 was（0.37±0.06）, and the expression quantity of YAP1 mRNA in siRNA-NC was（1.03±0.07）. The difference was statistically significant（P〈0.05）; Western blot showed that the expression quantity of YAP1 protein in siRNA-YAP1 was lower than that of siRNA-NC; the absorbance of YAP1 cells in siRNA-YAP1 was lower than that of siRNA-NC at 48, 72 and 96 h, and the difference was statistically significant（P〈0.05）. Conclusion The inhibitory effect of miR-200a on the proliferation of lung cancer cells is mainly achieved by targeting the YAP1 gene, which in turn plays a tumor suppressor function in lung cancer.
作者
谢悦
董礼文
王军
李雄伟
XIE Yue;DONG Liwen;WANG Jun;LI Xiongwei(Department of Cardiothoracic Surgery,Hangzhou Hospital of TCM,Affiliated Guangxing Hospital of Zhejiang Chinese Medicine University,Hangzhou 310000,China)
出处
《中国现代医生》
2018年第31期20-24,共5页
China Modern Doctor
基金
浙江省级公益技术应用研究计划(2016C33209)
