DNA高甲基化介导FOXO1转录表达下调促进鼻咽癌细胞增殖和侵袭能力  

Down-regulation of FOXO1 by DNA hypermethylation promoting the proliferation and invasion of nasopharyngeal carcinoma cells

作　　者：于鹏 金金欣 冀火金 黄兰川 刘进 蔡永林[4] YU Peng;YE Yinxin;JI Huojin;HUANG Lanchuan;LIU Jin;CAI Yonglin(School of Clinical Medicine,Guilin Medical University,Guilin 541199,China;Department of Clinical Laboratory,Wuzhou Red Cross Hospital,Wuzhou 543002,China;School of Public Health and Management,Youjiang Medical University For Nationalities,Baise 533000,China;Guangxi Health Commission Key Laboratory of Molecular Epidemiology of Nasopharyngeal Carcinoma,,Wuzhou 543002,China)

机构地区：[1]桂林医学院临床医学院,桂林541199 [2]梧州市红十字会医院医学检验科,梧州543002 [3]右江民族医学院公共卫生与管理学院,百色533000 [4]广西卫生健康委员会鼻咽癌分子流行病学重点实验室〔梧州市红十字会医院〕,梧州543002

出　　处：《广西医科大学学报》2024年第3期364-372,共9页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.81860601);广西自然科学基金资助项目(No.2020GXNSFAA297105)。

摘　　要：目的:探索叉头盒O1(FOXO1)在鼻咽癌(NPC)中异常表达的分子机制,及其对NPC细胞恶性生物学行为的影响。方法:采用GEO数据集分析NPC中FOXO1表达水平,采用R4.2.1软件进行基因集富集分析。构建FOXO1过表达的NPC细胞系5-8F、HONE1(FOXO1-5-8F、Ctrl-5-8F、FOXO1-HONE1、Ctrl-HONE1)。分别通过CCK-8法、划痕实验、Transwell侵袭实验检测细胞增殖、迁移和侵袭能力,流式细胞术分析细胞周期。通过重亚硫酸氢盐基因测序技术进行DNA甲基化测序。结果:FOXO1 mRNA在NPC细胞和组织中表达下调。FOXO1过表达组的细胞增殖能力和细胞迁移能力明显低于对照组(P<0.05),相同时间内,空白对照组的细胞侵袭数量高于FOXO1过表达组(P<0.05),FOXO1过表达抑制细胞周期(P<0.05)。在NPC组织中,FOXO1 DNA启动子区域存在甲基化位点,其甲基化水平明显高于非癌组织(P<0.05)。结论:NPC中FOXO1基因DNA高甲基化导致其转录下调,由此促进肿瘤细胞的增殖和侵袭。FOXO1可能是NPC的肿瘤抑制基因。Objective:To explore the molecular mechanism of abnormal expression of forkhead box O1(FOXO1)in nasopharyngeal carcinoma(NPC)and its impact on the malignant biological behavior of NPC cells.Methods:GEO dataset was used to analyze FOXO1 expression levels in NPC,and R4.2.1 software was used for gene set enrichment analysis.NPC cell lines 5-8F and HONE1 with overexpressing FOXO1(FOXO1-5-8F,Ctrl-5-8F,FOXO1 HONE1,Ctrl-HONE1)were constructed.Cell proliferation,migration,and invasion abilities were assessed through the cell counting kit-8(CCK-8)assay,scratch assay,and Transwell assay,respectively.The cell cycle was analyzed by using flow cytometry.FOXO1 DNA methylation sequencing by bisulfite gene sequencing technology was performed.Results:The expression of FOXO1 mRNA was down-regulated in NPC cells and tissues.Cell proliferation and migration abilities in the FOXO1 overexpression group were significantly lower than those in the control group(P<0.05),the number of cell invasion in the blank control group was higher than that in the FOXO1 overexpression group at the same time(P<0.05),and FOXO1 overexpression inhibited the cell cycle(P<0.05).The promoter region of FOXO1 DNA in NPC tissues was observed to contain a methylation site,with a significantly higher level of methylation compared to non-cancerous tissues(P<0.05).Conclusion:Down-regulation of FOXO1 transcription via DNA hypermethylation in NPC promotes the proliferation and invasion of NPC cells.FOXO1 may act as a tumor suppressor gene for NPC.

关 键 词：鼻咽癌 叉头盒O1 DNA甲基化 增殖 侵袭 

分 类 号：R739.6[医药卫生—肿瘤]