转化生长因子β调节子4对人肺鳞状细胞癌类器官增殖、侵袭和转移的影响  

Effect of transforming growth factorβregulator 4 on proliferation,invasion and metastasis of human lung squamous cell carcinoma organoids

作　　者：王安生[1] 王祖义[1] 郑传明[2] 陈娜娜[1] 李伟[3] 洪海宁 桑海威 王康武[1] 李其才 陈力维 朱浩楠 WANG Ansheng;WANG Zuyi;ZHENG Chuanming;CHEN Nana;LI Wei;HONG Haining;SANG Haiwei;WANG Kangwu;LI Qicai;CHEN Liwei;ZHU Haonan(Department of Thoracic Surgery,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,Anhui Province,China;Department of Emergency Surgery,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,Anhui Province,China;Department of Respiratory Diseases,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,Anhui Province,China;Graduate School of Bengbu Medical College,Bengbu 233030,Anhui Province,China;Department of Thoracic Surgery,Fuyang People′s Hospital,Fuyang 236012,Anhui Province,China)

机构地区：[1]蚌埠医学院第一附属医院胸外科,安徽蚌埠233004 [2]蚌埠医学院第一附属医院急诊外科,安徽蚌埠233004 [3]蚌埠医学院第一附属医院呼吸病科,安徽蚌埠233004 [4]蚌埠医学院研究生院,安徽蚌埠233030 [5]阜阳市人民医院胸外科,安徽阜阳236012

出　　处：《新乡医学院学报》2022年第10期907-911,918,共6页Journal of Xinxiang Medical University

基　　金：安徽省中央引导地方科技发展项目(编号:2020b07030008);蚌埠医学院转化医学重点专项项目(编号:BYTM2019019)。

摘　　要：目的探讨转化生长因子β调节子4(TBRG4)在肺癌细胞中的表达及其对人肺鳞状细胞癌类器官增殖、侵袭和转移的影响,并分析可能的分子机制。方法收集2020年1月至2021年1月蚌埠医学院第一附属医院手术切除的肺鳞状细胞癌患者的癌组织和癌旁组织,采用Western blot法检测癌组织和癌旁组织中TBRG4蛋白的表达。采用胰蛋白酶消化癌组织,分离肿瘤细胞,将分离的肿瘤细胞培养形成人肺鳞状细胞癌类器官。将人肺鳞状细胞癌类器官分为对照组和TBRG4沉默组,对照组类器官转染空载体慢病毒,TBRG4沉默组类器官转染TBRG4沉默慢病毒。采用细胞计数试剂盒-8检测人肺鳞状细胞癌类器官转染24、48、72、96 h时的增殖活力,Transwell法检测人肺鳞状细胞癌类器官的迁移和侵袭能力,Western blot法检测人肺鳞癌类器官中TBRG4、增殖细胞核抗体(PCNA)、磷酸酯酶与张力蛋白同源物(PTEN)、磷酸化磷脂酰肌醇激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)蛋白表达。结果肺鳞状细胞癌组织中TBRG4蛋白相对表达量显著高于癌旁组织(P<0.05)。TBRG4沉默组人肺鳞状细胞癌类器官转染24、48、72、96 h时的增殖活力均显著高于对照组(P<0.05)。TBRG4沉默组穿膜细胞数和迁移细胞数均显著少于对照组(P<0.05)。TBRG4沉默组人肺鳞状细胞癌类器官中TBRG4、PCNA、p-PI3K、p-AKT蛋白的相对表达量均显著低于对照组(P<0.05),PTEN蛋白的相对表达量显著高于对照组(P<0.05)。结论TBRG4在肺鳞状细胞癌组织中高表达,其可能是通过PTEN/PI3K/AKT信号通路调节人肺鳞状细胞癌类器官的增殖、侵袭和转移,TBRG4有望成为肺癌治疗的新靶点。Objective To investigate the expression of transforming growth factorβregulator 4(TBRG4)in lung cancer cells and its effect on proliferation,invasion and metastasis of human lung squamous cell carcinoma organoids,and to analyze the possible molecular mechanism.Methods The cancerous tissues and adjacent tissues removed from patients with human lung squamous cell carcinoma admitted at the First Affiliated Hospital of Bengbu Medical College from January 2020 to January 2021 were collected,and the expression of TBRG4 protein in cancerous tissues and adjacent tissues was detected by Western blot.Cancer tissue was digested with trypsin,and the tumor cells were isolated,then the isolated tumor cells were cultured to form human lung squamous cell carcinoma organoids.The human lung squamous cell carcinoma organoids divided into control group and TBRG4 silencing group.The organoids in the control group were transfected with empty vector lentivirus,the organoids in the TBRG4 silencing group were transfected with TBRG4 silencing lentivirus.The proliferation activity of human lung squamous cell carcinoma organoids was detected by cell counting kit-8,the migration and invasion ability of human lung squamous cell carcinoma organoids were detected by Transwell method,and the expressions of TBRG4,proliferating cell nuclear anlibody(PCNA),phosphatase and tensin homolog(PTEN),phosphorylated phosphatidylinositol-3-kinase(p-PI3K),phosphorylated protein kinase B(p-AKT)protein were detected by Western blot.Results The relative expression of TBRG4 protein in lung squamous cell carcinoma tissues was significantly higher than that in adjacent tissues(P<0.05).The proliferation activity of human lung squamous cell carcinoma organoids in the TBRG4 silencing group was significantly higher than that in the control group at 24,48,72,96 h after transfection.The number of transmembrane cells and the number of cell migration in the TBRG4 silencing group were significantly less than those in the control group(P<0.05).The relative expressions of TB

关 键 词：肺鳞状细胞癌 增殖 侵袭 转移 转化生长因子β调节子4 类器官 

分 类 号：R734.2[医药卫生—肿瘤]