苯并(a)芘诱导恶性转化的BEAS-2B细胞中miRNA和mRNA表达谱的改变  

Changes of mRNA and miRNA profiles in malignant transformation of BEAS-2B cells induced by benzo(a)pyrene

作　　者：栗振凯 李中秋 孟丽雅 张佳彤[2] 张巧 LI Zhenkai;LI Zhongqiu;MENG Liya;ZHANG Jiatong;ZHANG Qiao(Department of Toxicology,College of Public Health,Zhengzhou University,Zhengzhou 450001;Department of Disease Prevention and Control,Zhengzhou University Hospital,Zhengzhou 450001)

机构地区：[1]郑州大学公共卫生学院卫生毒理学教研室,郑州450001 [2]郑州大学医院疾病预防控制科,郑州450001

出　　处：《郑州大学学报（医学版）》2021年第6期762-767,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金项目(81172717)。

摘　　要：目的:探索与苯并(a)芘[B(a)P]诱导永生化人支气管上皮细胞(BEAS-2B细胞)恶性转化相关的miRNA和mRNA。方法:采用5 mg/L的B(a)P对BEAS-2B细胞进行染毒。取染毒和未染毒细胞,分别采用平板克隆形成实验、划痕实验、MTT法检测克隆形成能力、迁移能力和增殖活性;采用芯片技术检测miRNA和mRNA表达谱,以FC≥2.0且P<0.05为标准筛选出差异表达miRNA和mRNA,对其进行GO注释及KEGG通路分析。采用实时荧光定量PCR法检测两组细胞中hsa-miR-2115-3p和hsa-miR-4521的表达。结果:染毒组细胞克隆形成能力、迁移能力和增殖活性均明显增强(P<0.05)。芯片分析结果显示,染毒组和未染毒组细胞在不同的簇中;共筛选出127种差异表达miRNA(29种表达上调,98种表达下调)和159种差异表达mRNA(99种表达上调,60种表达下调)。GO注释及KEGG通路分析结果显示,差异表达mRNA可能参与了细胞活素-受体相互作用介导的信号通路、p53信号通路介导的细胞凋亡等。染毒组细胞中hsa-miR-2115-3p和hsa-miR-4521的表达低于未染毒组(P<0.05),与芯片结果一致。结论:筛选出B(a)P诱导BEAS-2B恶性转化相关的miRNA和mRNA。Aim:To explore the mRNA and miRNA associated with malignant transformation of immortalized human bronchial epithelial cells(BEAS-2B cells)induced by benzo(a)pyrene[B(a)P].Methods:BEAS-2B cells exposed to 5 mg/L B(a)P were the B(a)P group,and the cells without exposure were the control.The colony formation ability,migration ability and proliferation activity of the cells of the two groups were tested by plate colony formation test,scratch test and MTT assay;mRNA and miRNA profiles were detected using microarray techniques,the mRNA and miRNA with differential expression were screened by FC≥2.0 and P<0.05,and GO annotation and KEGG pathway analysis were performed.The expressions of hsa-miR-2115-3p and hsa-miR-4521 in the cells were detected by real-time fluorescent quantitative PCR.Results:The clony formation ability,migration ability and proliferation activity of the cells of B(a)P group were significantly enhanced(P<0.05).The cells of the two groups were in different clusters,and a total of 127 miRNAs(29 up-regulated and 98 down-regulated)and 159 mRNAs(99 up-regulated and 60 down-regulated)were screened.GO annotation and KEGG pathway analysis results showed that the differentially expressed mRNAs may be involved in the cytokine-receptor interaction signaling pathway and p53 signaling pathway,etc.The expressions of hsa-miR-2115-3p and hsa-miR-4521 in B(a)P group were lower those in the control group(P<0.05),and consistent with the microarray results.Conclusion:mRNA and miRNA associated with malignant transformation of BEAS-2B cells induced by B(a)P have been screened.

关 键 词：苯并(A)芘 BEAS-2B细胞 miRNA表达谱 mRNA表达谱 肺癌 

分 类 号：R114[医药卫生—卫生毒理学]