摘要
目的:对比缺氧环境下不同肺腺癌细胞株表达差异的microRNA,为进一步研究缺氧环境中肺癌细胞microRNA的作用机制提供参考。方法:建立缺氧模型,将培养于缺氧情况中及正常氧浓度下的GLC-82、A549细胞分别提取RNA,通过芯片检测及PCR验证,筛选差异表达的microRNA。利用TargetScanHuman 7.2数据库和cytoscape软件预测差异显著的microRNA分子调控的靶基因及可能参与的信号通路。结果:在缺氧条件下,A549细胞中3个microRNA表达上调,30个microRNA表达下调;而在GLC-82细胞中,2个microRNA表达上调,31个microRNA表达下调。两种细胞相比较,除miR-192-5p变化趋势相反外,其他microRNAs的表达变化趋势相同。通过KOBAS在线网站KEGG及GO预测分析,差异表达microRNA主要参与了肿瘤相关信号通路、胰岛素信号通路、Ras信号通路以及氧代谢等生物学过程。结论:肺癌细胞株在缺氧环境下和正常氧状态下mircoRNA出现表达差异,并且来源于不同遗传背景的细胞株之间也存在差异。差异表达的microRNA靶基因参与了多种细胞代谢相关通路,具有氧代谢,氧应答相关功能。
Objective:To investigate differentially expressed microRNAs between different lung cell lines under hypoxic conditions,and to provide a reference for further research on the mechanism of action of microRNAs in lung cancer cells under hypoxia.Methods:A model of hypoxia was established.RNA was extracted from GLC-82 and A549 cells cultured under a hypoxic or normal oxygen concentration,and microarray assay and PCR were used to screen out differentially expressed microRNAs.TargetScanHuman 7.2 and cytoscape software were used to predict the targeted genes regulated by differentially expressed microRNAs and the signaling path-ways involving these target genes.Results:Under the hypoxic condition,3 microRNAs were upregulated and 30 were downregulated in A549 cells,while 2 microRNAs were upregulated and 31 were downregulated in GLC-82 cells.miR-192-5 p showed an opposite change trend in these two cell lines,while the other microRNAs showed a similar change trend.Kyoto Encyclopedia of Genes and Genome and gene ontology analyses showed that the differentially expressed microRNAs were mainly involved in tumor-related signaling pathways,insulin signaling pathways,Ras signaling pathways,and oxygen metabolism.Conclusion:Differentially expressed microRNAs are found in lung cancer cell lines cultured under hypoxic or normal oxygen conditions,and there are also differentially expressed microRNAs between cells with different genetic backgrounds.The target genes of differentially expressed microRNAs are involved in a variety of cellular metabolic pathways and have the functions of oxygen metabolism and oxygen response.
作者
王莹
唐春明
李静林
邱丽娟
孙强明
Wang Ying;Tang Chunming;Li Jinglin;Qiu Lijuan;Sun Qiangming(Teaching and Research Section of Pathology,Basic Teaching Department,Haiyuan College,Kunming Medical University;Department of Oncology,The First People’s Hospital of Yunnan Province;Emergency Department,The First Affiliated Hospital of Kunming Medical University;Institute of Pediatric Diseases,Kunming Children’s Hospital;Molecular Epidemiology Joint Laboratory,Chinese Academy of Medical Sciences&Peking Union Medical College)
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2019年第7期891-897,共7页
Journal of Chongqing Medical University
基金
云南省教育厅科研资助项目(编号:2015Y530)