ALCAM基因敲除对苯并[a]芘恶性转化细胞THBEc1转录组的影响  

Effects of ALCAM gene knockout on transcriptome of benzo[a]pyrene-transformed human bronchial epithelial 16HBE cells

作　　者：李璐迪 陈洁 周川 马雪 付娟玲 姚碧云[1] 赵鹏[1] LI Lu-di;CHEN Jie;ZHOU Chuan;MA Xue;FU Juan-ling;YAO Bi-yun;ZHAO Peng(Beijing Key Laboratory of Toxicological Research and Risk Assessment for Food Safety,School of Public Health,Peking University Health Science Center,Beijing 100191,China)

机构地区：[1]北京大学公共卫生学院,食品安全毒理学研究与评价北京市重点实验室,北京100191

出　　处：《毒理学杂志》2021年第1期5-9,共5页Journal of Toxicology

基　　金：国家自然科学基金(81370079,81001253);北京市自然科学基金(7132122)。

摘　　要：目的检测活化白细胞黏附分子(ALCAM)基因敲除后苯并[a]芘(BaP)恶性转化细胞THBEc1转录组的改变,探究ALCAM在BaP致癌中的作用,为进一步阐明BaP致癌机制提供线索。方法选择两株ALCAM基因纯合敲除THBEc1细胞THBEc1-ΔALCAM-c5和THBEc1-ΔALCAM-c7及一株敲除对照细胞THBEc1-ctrl作为模型,利用转录组测序技术筛选细胞间差异表达基因,使用DAVID数据库对差异表达基因进行GO分析和KEGG通路富集分析。利用克隆形成实验检测ALCAM敲除对THBEc1细胞增殖的影响。结果转录组测序共发现336个基因在两株ALCAM敲除THBEc1细胞和THBEc1-ctrl细胞中表达差异在2倍以上(FDR<0.05),其中62个基因在两株ALCAM敲除THBEc1细胞中的表达低于THBEc1-ctrl细胞,274个基因在两株ALCAM敲除THBEc1细胞中的表达高于THBEc1-ctrl细胞。差异表达基因涉及多种生物学功能,主要包括细胞增殖、迁移、黏附、信号转导、凋亡、血管生成、炎症反应和免疫应答等。克隆形成实验证实ALCAM敲除可降低THBEc1细胞增殖能力。结论敲除ALCAM后BaP恶性转化细胞THBEc1转录组明显改变,增殖能力明显降低。ALCAM可能在BaP致癌中发挥重要作用,并可能作为BaP致癌的潜在生物标志物。Objective To explore the role of activated leukocyte cell adhesion molecule(ALCAM)in benzo[a]pyrene(BaP)-induced carcinogenesis through determining changes in transcriptomes of BaP-transformed human bronchial epithelial 16 HBE cells(THBEc1)after ALCAM gene knockout.Methods Two ALCAM knockout THBEc1 cell lines(THBEc1-ΔALCAM-c5 and THBEc1-ΔALCAM-c7)and one knockout control cell line(THBEc1-ctrl)were selected as cell models.Transcriptome sequencing technology was used to analyze and compare transcriptomes between ALCAM knockout THBEc1 cells and THBEc1-ctrl cells.Then biological information,including Gene Oncology and KEGG pathways,was gathered from DAVID database.Colony formation assay was used to determine the effects of ALCAM knockout on THBEc1 cells proliferation.Results A total of 336 genes were identified as differentially expressed genes via transcriptome sequencing analysis between ALCAM knockout THBEc1 cells and THBEc1-ctrl cells,among which 62 genes were down-regulated and 274 genes were up-regulated in ALCAM knockout THBEc1 cells.Differentially expressed genes were involved in many biological functions,including cell proliferation,migration,adhesion,signal transduction,apoptosis,angiogenesis,inflammation and immune response.Clone formation assay confirmed that ALCAM knockout inhibited the proliferation potential of THBEc1 cells.Conclusion ALCAM knockout led to significant changes in transcriptome of BaP-transformed cell THBEc1,and a decrease in proliferation potential.ALCAM might play an important role and act as a potential biomarker in BaP-induced carcinogenesis.

关 键 词：苯并[A]芘 ALCAM 转录组 THBEc1细胞 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]