Numb1与ITGB1的相互作用在胃癌恶化和耐药中的调控机制研究  

Study of the Regulatory Mechanism of the Interaction Between Numb1 and ITGB1 in Gastric Cancer Progression and Drug Resistance

作　　者：陈勇[1] 何冬雷[1] 周江浩 梁月祥 杨丞[1] CHEN Yong;HE Donglei;ZHOU Jianghao(The First Affiliated Hospital of Hainan Medical College,Hainan Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院,海南海口570102

出　　处：《河北医学》2022年第9期1420-1425,共6页Hebei Medicine

基　　金：海南省卫生健康行业科研项目,(编号:21A200052)。

摘　　要：目的:探讨衔接蛋白1(Numb1)与整合素β1(ITGB1)的相互作用对胃癌恶化和耐药的作用及其相关机制。方法:收集21例胃癌患者肿瘤及其癌旁组织,采用qRT-PCR检测Numb1和ITGB1的表达水平并统计两者表达水平相关性。通过慢病毒将过表达载体(flag-Numb1、flag-ITGB1和flag-NC)转染至BGC-823细胞,采用免疫共沉淀法检测Numb1和ITGB1的相互结合部位。将si-NC、si-Numb1和si-ITGB1转染至BGC-823细胞,实时定量PCR(qRT-PCR)法检测Numb1和ITGB1 mRNA表达水平,免疫印迹(Western blot,WB)法检测Numb1和ITGB1蛋白表达水平,CCK8检测细胞增殖水平和对奥沙利铂药物敏感性,细胞划痕实验检测细胞迁移能力,Transwell检测细胞侵袭能力。结果:与癌旁组织比较,肿瘤组织Numb1和ITGB1表达水平升高(P<0.05),Numb1与ITGB1表达水平成正相关(r=0.6270)。在细胞实验中,过表达flag-Numb1的flag免疫沉淀产物中可检测到ITGB1蛋白,过表达flag-ITGB1的flag免疫沉淀产物中可检测到Numb1蛋白。与si-NC组比较,si-Numb1组Numb1、ITGB1 mRNA和蛋白表达水平降低(P<0.05),细胞增殖能力减弱(P<0.05),对奥沙利铂药物敏感性增加(P<0.05),细胞迁移和侵袭能力降低(P<0.05)。与si-NC组比较,si-ITGB1组ITGB1 mRNA和蛋白表达水平降低(P<0.05),细胞增殖能力减弱(P<0.05),对奥沙利铂药物敏感性增加(P<0.05),细胞迁移和侵袭能力降低(P<0.05)。结论:Numb1通过与ITGB1相互作用调节ITGB1表达水平,促进胃癌细胞增殖、迁移、侵袭和对化疗药物耐药。Objective:To investigate the interaction between cohesive protein 1(Numb1)and integrinβ1(ITGB1)on gastric cancer progression and drug resistance and its related mechanisms.Methods:The expression levels of Numb1 and ITGB1 in tumors and adjacent tissues of 21 patients with gastric cancer were detected by qRT-PCR,and the correlation between the expression levels of Numb1 and ITGB1 was analyzed.The overexpressed vectors(flag-Numb1,flag-ITGB1,and flag-NC)were transfected into BGC-823 cells by lentivirus,and the binding sites of Numb1 and ITGB1 were detected by the immunoprecipitation method.si-NC,si-Numb1,and si-ITGB1 were transfected into BGC-823 cells.The mRNA expression levels of Numb1 and ITGB1 were detected by qRT-PCR and the protein expression levels of Numb1 and ITGB1 were detected by Western blot.CCK8 detected cell proliferation and sensitivity to oxaliplatin,cell migration was detected by cell scratch assay,and cell invasion was detected by Transwell.Results:The expression levels of Numb1 and ITGB1 were elevated in tumor tissues compared to paraneoplastic tissues(P<0.05),and Numb1 was positively correlated with ITGB1 expression levels(r=0.6270).In cellular assays,ITGB1 protein was detected in the immunoprecipitation products of flags overexpressing flag-Numb1,and Numb1 protein was detected in the immunoprecipitation products of flags overexpressing flag-ITGB1.Compared with the si-NC group,the si-Numb1 group showed reduced levels of Numb1 and ITGB1 mRNA and protein expression(P<0.05),reduced cell proliferation(P<0.05),increased sensitivity to oxaliplatin(P<0.05),and reduced cell migration and invasion(P<0.05).Compared with the si-NC group,the si-ITGB1 group showed reduced ITGB1 mRNA and protein expression levels(P<0.05),reduced cell proliferation(P<0.05),increased sensitivity to oxaliplatin(P<0.05)and reduced cell migration and invasion(P<0.05).Conclusion:Numb1 regulates ITGB1 expression levels through interaction with ITGB1 and promotes proliferation,migration,invasion and resistance to chemotherapeutic agen

关 键 词：胃癌 衔接蛋白1 整合素Β1 肿瘤耐药 

分 类 号：R735.2[医药卫生—肿瘤]