GOLM1调控PI3K/AKT/mTOR信号转导通路促进肺腺癌细胞增殖、侵袭和迁移的机制研究  

A study on mechanism of GOLM1 regulating PI3K/AKT/mTOR signaling pathway to promote proliferation,invasion and migration of lung adenocarcinoma cells

作　　者：朱海鹏[1] 胡军[1] 姜敏[2] 蔡若南 王俊巧 李莉[1] ZHU Haipeng;HU Jun;JIANG Min;CAI Ruonan;WANG Junqiao;LI Li(Department of Hematology and Oncology,Karamay Central Hospital,Karamay 834000,Xinjiang Uygur Autonomous Region,China;Department of Pathology,Karamay Central Hospital,Karamay 834000,Xinjiang Uygur Autonomous Region,China)

机构地区：[1]克拉玛依市中心医院血液肿瘤科,新疆克拉玛依834000 [2]克拉玛依市中心医院病理科,新疆克拉玛依834000

出　　处：《中国癌症杂志》2022年第3期207-217,共11页China Oncology

基　　金：新疆维吾尔自治区自然科学基金(2019D01A07)。

摘　　要：背景与目的:高尔基体膜蛋白1(Golgi membrane protein 1,GOLM1)在肺腺癌中发挥促癌作用,但对肺腺癌细胞增殖、侵袭和迁移的影响及其作用机制尚不明确。探究GOLM1对肺腺癌细胞增殖、侵袭和迁移的影响及其作用机制。方法:选取2019年4月—2021年4月在克拉玛依市中心医院行手术切除的肺腺癌患者的癌组织及相应癌旁组织标本各90例,采用免疫组织化学法检测肺腺癌组织和癌旁组织中GOLM1的表达,并分析肺腺癌组织中GOLM1表达与临床病理学特征的关系。采用蛋白质印迹法(Western blot)检测人肺上皮细胞系BEAS-2B及人肺腺癌H460、A549、PG49、H1299细胞系中GOLM1的表达。取对数生长期的肺腺癌A549细胞,随机分为空白组(细胞未转染)、GOLM1小干扰RNA阴性对照(small interfering RNA negative control,si-NC)组(细胞转染si-NC)、GOLM1小干扰RNA(GOLM1 small interfering RNA,si-GOLM1)组(细胞转染si-GOLM1)、胰岛素样生长因子-1(insulin-like growth factor-1,IGF-1)组[10μmol/L的磷脂酰肌醇3-激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)/哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号转导通路激活剂IGF-1处理30 min]和si-GOLM1+IGF-1组(10μmol/L的IGF-1处理30 min后再转染si-GOLM1),采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)法检测各组肺腺癌A549细胞增殖,采用划痕实验检测各组肺腺癌A549细胞迁移,采用transwell实验检测各组肺腺癌A549细胞侵袭,采用Western blot检测各组肺腺癌A549细胞中GOLM1、PI3K/AKT/mTOR信号转导通路相关蛋白的表达。BALB/c裸小鼠右侧皮下注射肺腺癌A549细胞建立移植瘤模型,分为空白组、si-NC组、si-GOLM1组、IGF-1组和si-GOLM1+IGF-1组,每组6只,注射6周后处死裸小鼠,收集肿瘤并测量肿瘤的质量和体积。结果:免疫组织化学结果显示,肺腺癌组织中GOLM1的表达阳性率显著高于癌旁组织(P<0.05)。GOLM1的表达与肿瘤分�Background and purpose:Golgi membrane protein 1(GOLM1)plays the role of an oncogene in lung adenocarcinoma(LUAD),however,the effects of GOLM1 on the proliferation,invasion and migration of LUAD cells and its mechanism are not clear yet.This study investigated the effects of GOLM1 on the proliferation,invasion and migration of LUAD cells and its mechanism of action.Methods:We selected cancer tissues and corresponding paracancerous tissue specimens from 90 LUAD patients who underwent surgical resection in Karamay Central Hospital from April 2019 to April 2021.The expression of GOLM1 protein in LUAD tissues and paracancerous tissues was detected by immunohistochemistry,and the relationship between GOLM1 protein expression and clinicopathological characteristics of LUAD tissues was analyzed.Western blot was used to detect the expression of GOLM1 protein in human lung epithelial cells BEAS-2B and human lung adenocarcinoma H460,A549,PG49 and H1299 cells.Lung adenocarcinoma A549 cells in logarithmic growth stage were randomly divided into blank group(cells not transfected),GOLM1 small interfering RNA negative control(si-NC)group(cells transfected with si-NC),GOLM1 small interfering RNA(si-GOLM1)group(cells transfected with si-GOLM1),insulin-like growth factor-1(IGF-1)group[10μmol/L phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)/mammalian target of rapamycin(mTOR)signaling pathway activator IGF-1 for 30 min]and si-GOLM1+IGF-1 group(after treatment with 10μmol/L IGF-1 for 30 min,si-GOLM1 was transfected).Cell counting kit-8 method was used to detect cell proliferation in each group of lung adenocarcinoma A549 cells.Scratch test was used to detect cell migration in each group of lung adenocarcinoma A549 cells.Transwell experiment was used to detect cell invasion in each group of lung adenocarcinoma A549 cells.Western blot was used to detect the expressions of GOLM1 and PI3K/AKT/mTOR signaling pathway related proteins in each group of lung adenocarcinoma A549 cells.Xenograft model was constructed by subcutaneou

关 键 词：高尔基体膜蛋白1 肺腺癌 细胞增殖 细胞侵袭 细胞迁移 磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白通路 

分 类 号：R734.2[医药卫生—肿瘤]