PM2.5对HBE细胞致癌致突变相关基因表达的影响

doi:10.3969/j.issn.1004-616x.2020.01.006

癌变·畸变·突变 ›› 2020, Vol. 32 ›› Issue (1): 33-38,42.doi: 10.3969/j.issn.1004-616x.2020.01.006

PM_2.5对HBE细胞致癌致突变相关基因表达的影响

王冰玉^1,2, 蔡颖^1,2, 郑凯^1,2, 谢红卫², 徐新云¹

1. 深圳市疾病预防控制中心环境与健康所, 广东深圳 518055;
2. 南华大学公共卫生学院, 湖南衡阳 421001

收稿日期:2019-08-07 修回日期:2019-11-01 出版日期:2020-01-31 发布日期:2020-02-05
通讯作者: 徐新云,E-mail:xyxu2008@163.com E-mail:xyxu2008@163.com
作者简介:王冰玉,E-mail:wbyfcfc@163.com。
基金资助:
深圳市科技研发基础研究项目（JCYJ20170413101713324）

Effect of PM_2.5 on expression of genes related to carcinogenesis and mutagenesis in HBE cells

WANG Bingyu^1,2, CAI Ying^1,2, ZHENG Kai^1,2, XIE Hongwei², XU Xinyun¹

1. Institute of Environment and Health, Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, Guangdong;
2. School of Public Health, University of South China, Hengyang 421001, Hunan, China

Received:2019-08-07 Revised:2019-11-01 Online:2020-01-31 Published:2020-02-05

摘要/Abstract

摘要： 目的：采用基因芯片技术与生物信息学分析方法，筛选PM_2.5染毒后的人支气管上皮细胞（HBE）与未染毒细胞比较的差异表达基因和通路，为进一步研究PM_2.5对HBE细胞致癌致突变作用的相关机制提供科学依据。方法：用50 μg/mL的PM_2.5水溶液处理HBE细胞，染毒24 h，用未染毒的细胞作为空白对照组，设立3组平行样品。提取RNA，荧光标记与纯化后进行芯片杂交，用Rosetta Resolver-System（Rosetta Biosoftware）进行数据前处理与统计分析，得到差异表达基因。将数据经过Cluster Profiler软件进行主成分和聚类分析、Pathway及GO（Gene Ontolog）分析，初步探讨PM_2.5染毒前后HBE细胞中基因的差异表达；通过String蛋白互作数据库分析差异表达基因的蛋白互作关系，筛选出节点数最多的核心蛋白。结果：根据预设的筛选条件，筛选出245个PM_2.5染毒前后HBE细胞中的差异表达基因，其中上调基因27个，下调基因218个，经过进一步分析，筛选出PHACTR2、SFRP1、WFDC1等排名前10的上调和下调的核心基因。排名前10的核心差异基因通过GO功能注释富集分析显示，差异表达基因主要富集在跨膜受体活性、受体活性、细胞信号传导、细胞分子传导等分子功能。同时富集于离子跨膜转运、细胞对脂多糖无机离子跨膜转运、细胞营养转运等生物过程；排名前10的差异表达基因通过KEGG富集分析显示，差异表达基因主要富集在涉及肿瘤细胞迁移、胆汁代谢相关、神经活性、遗传毒性等方面的7个核心通路；蛋白互作用网络图筛选出SST、BDNF、NCAM1、SSTR1和Bcl2L11等5个核心蛋白。结论：PM_2.5可引起HBE细胞致癌致突变相关基因的差异表达，可为进一步研究PM_2.5的致癌致突变作用机制提供科学依据。

关键词: 细颗粒物, 人支气管上皮细胞, 基因芯片, 生物学功能注释, KEGG富集分析

Abstract: OBJECTIVE: To use gene chip technology and bioinformatics to investigate PM_2.5-exposure on expression of genes related to carcinogenesis and mutagenesis in human bronchial epithelial (HBE) cells. METHODS: HBE cells were treated with 50 μg/mL water-soluble PM_2.5 for 24 h. Untreated cells were used as a blank control group,and three parallel samples were established. Extracted RNA samples were subjected to fluorescent-labeling,purification and hybridization to the chip. Collected data were pre-processed and statistically calculated using the Rosetta Resolver^® System (Rosetta Biosoftware) to analyze differentially expressed genes. The data were analyzed using the Cluster Profiler software for principal component and cluster analysis,and pathway and GO (Gene Ontolog) analysis. Differences in gene expression of HBE cells after PM_2.5 exposure were preliminarily investigated. Protein interaction relationships of differentially expressed genes were analyzed using the String protein interaction database,then the core protein with the largest number of nodes was selected. RESULTS: According to the preset screening conditions,245 differentially expressed genes were screened,including 27 up-regulated genes and 218 down-regulated genes. After further analyses,the top 10 up-regulated and down-regulated core genes such as PHACTR2,SFRP1,WFDC1 were screened out. The top 10 core differential gene by GO functional annotation enrichment analysis show that the differentially expressed genes were mainly enriched in molecular functions such as transmembrane receptor activity,receptor activity,cell signaling,and cell molecular conduction. Their mechanisms involved biological transmembrane transport,translocation of cells to lipopolysaccharide inorganic ions,and transport of cellular nutrients. The KEGG enrichment analyses show that the differentially expressed genes were mainly enriched in seven core pathways involved in tumor cell migration,bile metabolism related,neural activity,and genotoxicity. Protein interaction networking mapped out 5 core proteins including SST,BDNF,NCAM1,SSTR1 and Bcl2L11. CONCLUSION: Exposure of HBE cells to PM_2.5 showed differential expression of genes which are involved with carcinogenesis and mutagenesis. The information is useful for better understanding the carcinogenic effect of PM_2.5.

Key words: PM_2.5, human bronchial epithelial cells, gene chip, Gene Ontolog, KEGG enrichment analysis

中图分类号:

R122.7

王冰玉, 蔡颖, 郑凯, 谢红卫, 徐新云. PM_2.5对HBE细胞致癌致突变相关基因表达的影响[J]. 癌变·畸变·突变, 2020, 32(1): 33-38,42.

WANG Bingyu, CAI Ying, ZHENG Kai, XIE Hongwei, XU Xinyun. Effect of PM_2.5 on expression of genes related to carcinogenesis and mutagenesis in HBE cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(1): 33-38,42.

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PM_2.5对HBE细胞致癌致突变相关基因表达的影响

Effect of PM_2.5 on expression of genes related to carcinogenesis and mutagenesis in HBE cells

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PM2.5对HBE细胞致癌致突变相关基因表达的影响

Effect of PM2.5 on expression of genes related to carcinogenesis and mutagenesis in HBE cells

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PM_2.5对HBE细胞致癌致突变相关基因表达的影响

Effect of PM_2.5 on expression of genes related to carcinogenesis and mutagenesis in HBE cells