G蛋白β2亚基对结直肠癌细胞转移能力的影响  

Effect of G protein subunit beta 2 on the metastasis ability of colorectal cancer cells

作　　者：刘荣 王永霞[1] LIU Rong;WANG Yongxia(Department of Pathology,School of Basic Medical Sciences,Xinxiang Medical University,Xinxiang 453003,Henan Province,China)

机构地区：[1]新乡医学院基础医学院病理学系,河南新乡453003

出　　处：《新乡医学院学报》2024年第7期619-624,630,共7页Journal of Xinxiang Medical University

基　　金：国家自然科学基金资助项目(编号:81702850);河南省科技攻关项目(编号:212102310145)。

摘　　要：目的探讨G蛋白β2亚基(GNB2)对结直肠癌细胞体外迁移和体内转移能力的影响及机制。方法将对数生长期人胚肾细胞293FT随机分为对照组和shGNB2组,对照组293FT细胞转染PSPAX2、PMD2G、Control质粒,shGNB2组293FT细胞转染PSPAX2、PMD2G、shGNB2质粒,分别收集病毒上清液。将对数生长期人结直肠癌细胞HCT116、RKO按随机数字表法分为对照组和shGNB2组,应用293FT细胞对照组病毒上清液转染对照组HCT116和RKO细胞,应用293FT细胞shGNB2组病毒上清液转染shGNB2组HCT116和RKO细胞,应用实时荧光定量聚合酶链反应法检测对照组和shGNB2组HCT116、RKO细胞中GNB2 mRNA的表达,Western blot法检测对照组和shGNB2组HCT116、RKO细胞中GNB2、波形蛋白(Vimentin)、神经钙黏蛋白(N-cadherin)和E-钙黏蛋白(E-cadherin)的表达,划痕愈合实验检测对照组和shGNB2组HCT116、RKO细胞划痕愈合率,Transwell小室实验检测对照组和shGNB2组HCT116、RKO细胞迁移细胞数。分别取对照组和shGNB2组HCT116细胞2×106个注射于裸鼠皮下,3周后取出皮下肿瘤,剪成1 mm 3大小的组织块。按随机数字表法将8只4~5周龄雌性裸鼠分为对照组和shGNB2组,每组4只;将对照组HCT116细胞皮下瘤组织块接种于对照组裸鼠回盲部肠浆膜处,shGNB2组HCT116细胞皮下瘤组织块接种于shGNB2组裸鼠回盲部肠浆膜处;记录裸鼠死亡时间,取出肝脏,观察肝脏肿瘤转移数。结果shGNB2组HCT116、RKO细胞中GNB2 mRNA和蛋白相对表达量显著低于对照组(P<0.01)。培养48 h时,shGNB2组HCT116、RKO细胞划痕愈合率显著低于对照组(P<0.01),shGNB2组HCT116、RKO细胞迁移数显著少于对照组(P<0.01)。shGNB2组裸鼠肝内转移瘤数显著少于对照组(P<0.05)。shGNB2组HCT116、RKO细胞中Vimentin、N-cadherin蛋白相对表达量显著低于对照组,E-cadherin蛋白相对表达量显著高于对照组(P<0.01)。结论干扰GNB2表达可有效抑制结直肠癌细胞的转移能力,其机制可能是GNObjective To investigate the effect and mechanism of G protein subunit beta 2(GNB2)on the migration and metastasis of colorectal cancer cells.Methods The human embryonic kidney cells 293FT in the logarithmic growth phase were randomly divided into the control group and the shGNB2 group.The 293FT cells in the control group were transfected with PSPAX2,PMD2G,and Control plasmids,while the 293FT cells in the shGNB2 group were transfected with PSPAX2,PMD2G,and shGNB2 plasmids;and the viral supernatants were collected,respectively.Human colorectal cancer cells HCT116 and RKO in the logarithmic growth phase were divided into the control group and the shGNB2 group according to the random number table method.HCT116 and RKO cells in the control group were transfected with the viral supernatants from the control group of 293FT cells;HCT116 and RKO cells in the shGNB2 group were transfected with the viral supernatants from the shGNB2 group of 293FT cells.The expression of GNB2 mRNA in HCT116 and RKO cells in the control group and the shGNB2 group was detected by real-time fluorescence quantitative polymerase chain reaction.The expressions of GNB2,Vimentin,N-cadherin and E-cadherin proteins in HCT116 and RKO cells in the control group and the shGNB2 group were detected by Western blot.The wound-healing rates of HCT116 and RKO cells in the control group and the shGNB2 group were measured by wound-healing assay.The migration of HCT116 and RKO cells in the control group and the shGNB2 group was detected by Transwell assay.A total of 2×106 HCT116 cells were taken from the control group and the shGNB2 group,respectively;and which were injected subcutaneously in nude mice,and the subcutaneous tumors were removed after 3 weeks and cut into small tissue pieces of 1 mm 3.Eight female nude mice aged 4-5 weeks were divided into the control group and the shGNB2 group according to the random number table method,with 4 mice in each group.The tissue pieces of subcutaneous tumors from HCT116 cells in the control group were inoculated in th

关 键 词：结直肠癌 G蛋白β2亚基 迁移 转移 

分 类 号：R735.3[医药卫生—肿瘤]