DNMT3a在肺腺癌中的表达及癌细胞迁移的影响分析

DNMT3a promotes the migration of lung adenocarcinoma cells by up-regulating Snail

目的探讨DNA甲基转移酶3a(DNMT3a)在肺腺癌中的表达水平及影响肺腺癌细胞迁移的分子机制。方法回顾性分析2023年1~12月在安徽医科大学第一附属医院胸外科接受手术治疗的41例肺腺癌患者临床资料,使用IHC和蛋白质印迹实验(WB)检测并分析DNMT3a在41例患者肺腺癌组织及癌旁组织中的表达水平;通过分析TCGA数据库揭示DNMT3a与肺腺癌患者预后关系。慢病毒介导SK-LU-1和A549细胞系过表达或敲低DNMT3a,通过IHC、WB、实时荧光定量聚合链反应(qRT-PCR)、DNMT3a抑制剂实验观察DNMT3a对蜗牛家族转录因子(Snail)表达的影响;细胞划痕实验和Transwell细胞迁移实验检测过表达或敲低DNMT3a对肺腺癌细胞迁移的影响。结果DNMT3a高表达对比DNMT3a低表达的肺腺癌患者生存期较短,差异有统计学意义(P<0.01);DNMT3a和Snail在肺腺癌组织中较癌旁组织相比均高表达(P<0.01),且DNMT3a和Snail在肺腺癌组织中的表达呈正相关(r=0.612,P<0.01)。与对照组相比,过表达DNMT3a的SK-LU-1细胞中Snail表达上调并增加了肺腺癌细胞的迁移数量(P<0.01),敲低DNMT3a的A549细胞中Snail表达下调并减少了肺腺癌细胞的迁移数量(P<0.01)。结论DNMT3a在肺腺癌组织中高表达且与不良预后显著相关,DNMT3a通过上调Snail的表达促进肺腺癌细胞的迁移。

Objective The aim of the study was to investigate the expression of DNMT3a and the molecular mechanism of DNMT3a in lung adenocarcinoma.Methods The expression levels of DNMT3a in 41 pairs of lung adenocarcinoma tissues and adjacent tissues were detected and analyzed by IHC and Western blot in the First Affiliated Hospital of Anhui Medical University From January 2023 to December 2023.TCGA database was used to analyze the relationship between DNMT3a expression level and the prognosis of patients with lung adenocarcinoma.SK-LU-1 and A549 cells were transfected with lentivirus to over-express or knockdown DNMT3a expression,and the effect of DNMT3a on Snail expression were observed by IHC,Western blot,qRT-PCR,and DNMT3a inhibitor experiments.The effect of over-expression or knockdown of DNMT3a on the migration of lung adenocarcinoma cells was detected by Wound healing assay and Transwell cell migration assay.Results High expression of DNMT3a was significantly associated with poor prognosis of lung adenocarcinoma patients(P<0.01).Compared with adjacent tissues,DNMT3a and Snail were both highly expressed in lung adenocarcinoma tissues(P<0.01),and the expression of DNMT3a and Snail was remarkably correlated in lung adenocarcinoma tissues through Mann-Kendall test(correlation coefficient=0.612,P<0.01).The prognosis of lung adenocarcinoma patients with high DNMT3a expression was poor(P<0.01).Compared with the control group,DNMT3a over-expression up-regulated the expression of Snail and promoted the migration in SK-LU-1 cells(P<0.01),while DNMT3a knockdown down-regulated the expression of Snail and inhibited the migration in A549 cells(P<0.01).Conclusion DNMT3a is highly expressed in human lung adenocarcinoma tissues and is significantly associated with poor prognosis.DNMT3a promotes the migration of lung adenocarcinoma cells through up-regulating the expression of Snail.