甲基丙烯酸环氧丙酯诱导16HBE细胞恶性转化过程中LncRNA表达特征分析及ceRNA调控网络预测

doi:10.3969/j.issn.1004-616x.2022.01.001

癌变·畸变·突变 ›› 2022, Vol. 34 ›› Issue (1): 1-6.doi: 10.3969/j.issn.1004-616x.2022.01.001

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甲基丙烯酸环氧丙酯诱导16HBE细胞恶性转化过程中LncRNA表达特征分析及ceRNA调控网络预测

李昕苇^1,2, 王全凯^1,2, 马顺鹏^1,2, 王苗^1,2, 乌瀚宝栎尔¹, 顾轶婷¹, 康同影¹, 许建宁^1,2

1. 中国疾病预防控制中心职业卫生与中毒控制所, 北京 100050;
2. 中国疾病预防控制中心化学污染与健康安全重点实验室, 北京 100050

收稿日期:2021-08-20 修回日期:2021-10-14 出版日期:2022-01-31 发布日期:2022-02-15
通讯作者: 许建宁,E-mail:jnx999@263.net E-mail:jnx999@263.net
作者简介:李昕苇,E-mail:erislee1996@163.com。
基金资助:
国家自然科学基金（81673221）

Long non-coding RNA expression analysis and ceRNA regulatory network in glycidyl methacrylate-induced malignant transformation of 16HBE cells

LI Xinwei^1,2, WANG Quankai^1,2, MA Shunpeng^1,2, WANG Miao^1,2, WUHAN Baolier¹, GU Yiting¹, KANG Tongying¹, XU Jianning^1,2

1. National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing 100050;
2. Key Laboratory of Chemical Safety and Health, Chinese Center for Disease Control and Prevention, Beijing 100050, China

Received:2021-08-20 Revised:2021-10-14 Online:2022-01-31 Published:2022-02-15

摘要/Abstract

摘要： 目的： 筛选甲基丙烯酸环氧丙酯（GMA）诱导人支气管上皮16HBE细胞恶性转化过程中不同时期差异表达的LncRNA并以此为基础构建ceRNA调控网络，分析相关信号通路和生物学功能。方法： 收集用GMA和溶剂DMSO分别处理的16HBE细胞。采用LncRNA芯片检测并分析两组细胞中LncRNA表达量的差异，运用TargetScan和MiRanda数据库筛选出关键的差异表达LncRNA，使用Cytoscape构建以差异表达LncRNA为核心的ceRNA调控网络，之后对差异表达LncRNA的靶基因和通路进行分析预测，通过实时荧光定量PCR （qPCR）对上述LncRNA相对表达量进行检测验证。结果： 芯片筛选结果发现，与DMSO对照组相比，GMA处理组共16个LncRNA在细胞恶性转化过程中表达上调。在此基础上筛选构建了由3个LncRNA，32个mRNA和204个miRNA组成的LncRNA相关ceRNA调控网络，其中有3个调控轴（LncRNA G013234-hsa-miR-378b-TMEM129、LncRNA CTA-384D8.35-hsa-miR-486-3p-LYPD、LncRNA G087116-miR-29b-3p-NAV1）与GMA诱导16HBE细胞恶性转化过程相关（P<0.05），提示这3个调控轴是与此恶性转化过程关联较为密切的ceRNA三元组。qPCR验证结果表明在细胞恶性转化过程的不同时期LncRNA G013234，CTA-384D8.35和G087116的相对表达水平变化趋势与LncRNA芯片结果一致。结论： 在GMA诱导16HBE细胞发生恶性转化过程中，LncRNA G013234、CTA-384D8.35、G087116在恶性转化不同时期均发生上调，以此为基础构建的ceRNA调控网络及预测得到细胞恶性转化相关的关键mRNA，为GMA诱导16HBE细胞恶性转化机制研究提供了支持数据。

关键词: 甲基丙烯酸环氧丙酯, 细胞恶性转化, 长链非编码RNA, ceRNA网络

Abstract: OBJECTIVE: To investigate differential expression of long non-coding RNA (LncRNA) during glycidyl methacrylate (GMS)-induced malignant transformation of human bronchial epithelial 16HBE cells,and to explore the regulatory effect of LncRNA on cell function and related signaling pathways at different stages of malignant transformation. METHODS: 16HBE cells were divided into GMA treatment group and DMSO control group,and the cells of the two groups were collected respectively. LncRNA microarray was used to detect and analyze differences of LncRNA expression in the malignantly transformed 16HBE cells. The key differentially expressed LncRNAs were screened out using TargetScan and MiRanda databases. Cytoscape was used to construct a ceRNA regulatory network with differentially expressed LncRNAs as the core. After that,the target genes and pathways of differentially expressed LncRNA were analyzed and predicted,and the relative expression levels of the above LncRNA were detected and verified by real-time fluorescence quantitative PCR. RESULTS: Microarray screening results showed that,compared with DMSO group,16 LncRNAs in GMA group were up-regulated in the process of the malignant transformation. On this basis,a LncRNA-related ceRNA regulatory network consisting of 3 LncRNAs,32 mRNAs and 204 miRNAs was constructed. Three regulatory axes in the network were significantly correlated with GMA-induced malignant transformation of 16HBE cells (P<0.05),and this showed that LncRNA G013234-hsa-miR-378b-TMEM129,LncRNA CTA-384D8.35-hsa-miR-486-3p-LYPD,LncRNA G087116-miR-29b-3p-NAV1 were closely associated with the malignant transformation process ceRNA triples. Results of qPCR showed that relative expression levels of LncRNA G013234,CTA-384D8.35 and G087116 in different cell stages were consistent with results of LncRNA microarray. CONCLUSION: During GMA-induced malignant transformation of 16HBE cells,LncRNA G013234,CTA-384D8.35 and G087116 were up-regulated at different stages of transformation. Construction of the ceRNA regulatory network and key mRNAs related to cell malignant transformation formed the theoretical basis for the process of GMA-induced malignant transformation of 16HBE cells.

Key words: glycidyl methacrylate, malignant transformation, LncRNA, ceRNA network

中图分类号:

R994.6

李昕苇, 王全凯, 马顺鹏, 王苗, 乌瀚宝栎尔, 顾轶婷, 康同影, 许建宁. 甲基丙烯酸环氧丙酯诱导16HBE细胞恶性转化过程中LncRNA表达特征分析及ceRNA调控网络预测[J]. 癌变·畸变·突变, 2022, 34(1): 1-6.

LI Xinwei, WANG Quankai, MA Shunpeng, WANG Miao, WUHAN Baolier, GU Yiting, KANG Tongying, XU Jianning. Long non-coding RNA expression analysis and ceRNA regulatory network in glycidyl methacrylate-induced malignant transformation of 16HBE cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2022, 34(1): 1-6.

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甲基丙烯酸环氧丙酯诱导16HBE细胞恶性转化过程中LncRNA表达特征分析及ceRNA调控网络预测

Long non-coding RNA expression analysis and ceRNA regulatory network in glycidyl methacrylate-induced malignant transformation of 16HBE cells

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