YTHDF1对肺腺癌细胞增殖和迁移能力的影响  

Regulatory effects of YTHDF1 on the proliferation and migration of lung adenocarcinoma cells

作　　者：岳洪娟[1] 王丽红[1] 王晓雨 马俊玲 刘婷 王嘉[1] Yue Hongjuan;Wang Lihong;Wang Xiaoyu;Ma Junling;Liu Ting;Wang Jia(Department of Infectious Diseases,the First Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Endoscopy Center,the First Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第一医院感染性疾病科,石家庄050000 [2]河北医科大学第一医院内镜中心,石家庄050000

出　　处：《国际呼吸杂志》2023年第7期810-815,共6页International Journal of Respiration

基　　金：河北省政府资助临床医学优秀人才培养项目(LS202008、LS202212);河北省重点研发计划项目(223777113D);河北省医学科学研究课题计划(20210236、20231019)。

摘　　要：目的研究YTH结构域N6-甲基腺苷RNA结合蛋白1(YTHDF1)对肺腺癌细胞增殖和迁移能力的影响。方法本研究为实验研究。选择2022年4月至12月期间于河北医科大学第一医院胸外科进行手术治疗的6例肺腺癌患者为研究对象,收集手术过程中切除的肺腺癌组织和癌旁正常肺组织。通过检索TCGA数据库和GEO数据库分析YTHDF1 mRNA在肺腺癌组织和正常肺组织中的表达水平,免疫组织化学染色对6例肺腺癌患者的肺腺癌组织及其配对的癌旁正常肺组织中YTHDF1的细胞定位进行检测。应用shRNA转染构建敲低YTHDF1的肺腺癌细胞株NCI-H1975,通过实时定量反转录PCR和蛋白质印迹法验证YTHDF1的敲低效率。采用CCK-8实验和Transwell迁移实验明确敲低YTHDF1对肺腺癌细胞增殖和迁移能力的影响。结果TCGA数据库和GEO数据库提示YTHDF1 mRNA在肺腺癌组织中的表达水平均高于正常肺组织[(6.36±0.54)比(5.71±0.31)、(9.83±0.26)比(9.69±0.07)],差异均有统计学意义(均P<0.01),YTHDF1主要表达于细胞质。YTHDF1 mRNA、YTHDF1在敲低组的表达水平均低于对照组[(0.46±0.08)比(1.03±0.07)、(0.48±0.09)比(0.86±0.53)],差异均有统计学意义(均P<0.001),本研究成功构建了敲低YTHDF1的肺腺癌细胞株。CCK-8实验表明,培养48 h、72 h和96 h后,敲低组的增殖能力均低于对照组[(0.22±0.01)比(0.24±0.01)、(0.26±0.01)比(0.32±0.01)、(0.32±0.01)比(0.43±0.01),均P<0.05]。Transwell迁移实验表明,敲低组迁移的肺腺癌细胞数量低于对照组[(299.33±25.50)个比(504.67±36.12)个,P=0.001]。结论敲低YTHDF1可以抑制肺腺癌细胞的增殖和迁移能力。Objective To detect the regulatory effect of YTH N6-methyladenosine RNA binding protein 1(YTHDF1)on the proliferation and migration abilities of lung adenocarcinoma cells.Methods It was an experimental study.Six patients with lung adenocarcinoma who underwent surgical treatment in the Department of Thoracic Surgery,the First Hospital of Hebei Medical University between April and December 2022 were selected as the research subjects,and the lung adenocarcinoma tissues and adjacent normal lung tissues resected during the operation were collected.The expression levels of YTHDF1 mRNA in lung adenocarcinomas and normal lung tissues were analyzed by searching The Cancer Genome Atlas(TCGA)database and Gene Expression Omnibus(GEO)database.Immunohistochemical staining was performed to detect the subcellular localization of YTHDF1 in lung adenocarcinomas and their paired adjacent normal lung tissues of 6 patients with lung adenocarcinoma.The lung adenocarcinoma cell line NCI-H1975 with YTHDF1 knockdown was constructed by short-hairpin RNA(shRNA)transfection,and the transfection efficiency was verified by real-time quantitative reverse transcription PCR and Western blot analysis.Cell proliferation and migration in NCI-H1975 cells with YTHDF1 knockdown were assessed by Cell Counting Kit-8(CCK-8)assay and Transwell migration assay,respectively.Results Both TCGA([6.36±0.54]vs[5.71±0.31])and GEO databases([9.83±0.26]vs[9.69±0.07])indicated that the mRNA level of YTHDF1 in lung adenocarcinoma tissues was significantly higher than that in normal lung tissues(both P<0.01),and YTHDF1 was mainly expressed in the cytoplasm.The mRNA([0.46±0.08]vs[1.03±0.07])and protein levels([0.48±0.09]vs[0.86±0.53])of YTHDF1 in NCI-H1975 with YTHDF1 knockdown were significantly lower than those of negative controls(both P<0.001),suggesting the successful construction of a lung adenocarcinoma cell line with YTHDF1 knockdown.CCK-8 assay showed that the proliferative rate of NCI-H1975 with YTHDF1 knockdown at 48 h([0.22±0.01]vs[0.24±0.01]),72

关 键 词：肺腺癌 N6-甲基腺苷 YTH结构域N6-甲基腺苷RNA结合蛋白1 增殖 迁移 

分 类 号：R734.2[医药卫生—肿瘤]