摘要
目的在生物信息学基础上探讨微小RNA(miR)-140-3p靶向细胞分裂周期相关蛋白8(CDCA8)抑制肺腺癌细胞的侵袭和转移。方法通过GEO数据库中的GEO2R分析肺腺癌芯片数据中差异表达的miRNA。Target Scan Human7.2和miRWalk数据库查找miR-140-3p的靶基因。Cytoscape3.7.2软件筛选出Hub基因。GEPIA数据库查询靶基因在肺腺癌组织和正常肺组织中的表达水平,在肺腺癌不同分期中的表达水平以及靶基因的表达水平与肺腺癌患者总体生存率的关系。Star Base数据库查找miR-140-3p在肺腺癌中的生存分析及与CDCA8的表达水平在肺腺癌中的相关性。Real-time PCR检测miR-140-3p在正常肺上皮细胞BEAS-2B和肺腺癌细胞A549的表达水平以及感染效率。Real-time PCR和Western blotting实验检测过表达miR-140-3p后CDCA8 m RNA和蛋白的表达水平。双荧光素酶报告基因实验验证miR-140-3p是否与CDCA8直接结合。Transwell侵袭实验检测过表达miR-140-3p和CDCA8对肺腺癌细胞侵袭力的影响。结果通过GEO等数据库的分析结果显示,miR-140-3p在正常肺组织中的表达水平明显高于在肺腺癌中的表达水平,其预测靶基因CDCA8在肺腺癌中的表达水平明显高于在正常肺组织中的表达水平,且CDCA8与miR-140-3p的表达水平在肺腺癌中成负相关。实验结果显示,miR-140-3p在A549细胞中的表达明显低于在BEAS-2B细胞中的表达(P<0.05);用过表达慢病毒感染细胞后miR-140-3p的表达水平明显升高(P<0.05);过表达miR-140-3p后CDCA8的m RNA和蛋白表达水平明显下调(P<0.05);双荧光素酶报告基因实验结果显示,miR-140-3p能与CDCA8直接结合(P<0.05);与对照组相比,过表达miR-140-3p可抑制肺腺癌细胞A549的侵袭转移,而CDCA8可逆转miR-140-3p对肺腺癌细胞A549侵袭力的抑制作用(P<0.05)。结论MiR-140-3p靶向CDCA8抑制肺腺癌细胞的侵袭转移。
Objective To investigate the effect of micro RNA(miR)-140-3p targeting cell division cycle associated 8(CDCA8)on invasion and metastasis of lung adenocarcinoma cells.Methods The differentially expressed miRNAs were analyzed by GEO2R in GEO database.The target genes of miR-140-3p were searched by Target Scan human7.2 and miRWalk databases.The hub gene was screened by Cytoscape 3.7.2 software.GEPIA database was used to query the expression levels of target gene in lung adenocarcinoma tissues and normal lung tissues,the expression levels in different stages of lung adenocarcinoma,and the relationship between the expression levels of target gene and the overall survival rate of lung adenocarcinoma patients.The survival analysis of miR-140-3p in lung adenocarcinoma and the correlation between miR-140-3p and CDCA8 expression levels were searched in star Base database.Real-time PCR was used to detect the expression levels of miR-140-3p in normal lung epithelial cells BEAS-2 B and lung adenocarcinoma cells A549,as well as the efficiency of infection.Expression levels of CDCA8 m RNA and protein were detected by Real-time PCR and Western blotting experiments after overexpression of miR-140-3p.Dual-luciferase reporter assay verified whether miR-140-3p directly binds to CDCA8.Transwell invasion assay detected the effect of overexpression of miR-140-3p and CDCA8 on the invasiveness of lung adenocarcinoma cells.Results Analysis result from GEO and other databases showed that the expression level of miR-140-3p in normal lung tissues was significantly higher than that in lung adenocarcinoma,and its predicted target gene CDCA8 expression level in lung adenocarcinoma was significantly higher than that in normal lung tissues,and CDCA8 was negatively correlated with the expression level of miR-140-3p in lung adenocarcinoma.The experimental result showed that the expression of miR-140-3p in A549 cells was significantly lower than that in BEAS-2 B cells(P<0.05).The expression level of miR-140-3p increased significantly after lentivira
作者
郑荃
胡雅琼
白俊
尹崇高
李洪利
刘雨清
ZHENG Quan;HU Ya-qiong;BAI Jun;YIN Chong-gao;LI Hong-li;LIU Yu-qing(Department of Pathology,Weifang Medical University,Shandong Weifang 261053,China;College of Nursing,Weifang Medical University,Shandong Weifang 261053,China;Medical Research Center,Weifang Medical University,Shandong Weifang 261053,China)
出处
《解剖学报》
CAS
CSCD
北大核心
2021年第4期589-600,共12页
Acta Anatomica Sinica
基金
国家自然科学基金(81702932,81641111)
山东省自然科学基金(ZR2019MH033)
潍坊市科学技术发展计划项目(2018GX077)
山东省高等学校青创人才引育计划(205)。
