抗原加工相关的转运蛋白1在胶质瘤中的表达及临床意义  

Transporter associated with antigen processing 1 functions as a tumor suppressor in glioma

作　　者：陈程 李军亮 许新科 陈伟 李方成 Chen Cheng;Li Junliang;Xu Xinke;Chen Wei;Li Fangcheng(Department of Neurosurgery,Guangzhou Women and Children’s Medical Center,Guangzhou 510623,China)

机构地区：[1]广州医科大学附属广州市妇女儿童医疗中心,510623

出　　处：《中华实验外科杂志》2020年第4期680-684,共5页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81072081、81272774、81572497)。

摘　　要：目的探讨抗原加工相关的转运蛋白1(TAP1)是主要组织相容性复合体-Ⅰ(MHC-Ⅰ)类抗原呈递途径所必需,在肿瘤免疫学监测重要标志性作用。方法通过慢病毒转导3种胶质瘤细胞株(U87、U251和GL261)中过表达和敲低TAP1的表达水平,实验分为正常对照组、慢病毒过表达空载体对照组、慢病毒敲除空载体对照组、TAP1过表达组和敲除组;蛋白质印迹法(Western blot)和定量聚合酶链反应(qPCR)检测TAP1和MHC-Ⅰ的表达和相互关系;使用Transwell系统检测细胞增殖和侵袭功能;流式细胞检测MHC-Ⅰ在TAP1过表达和敲低的GL261细胞表面的表达;免疫组织化学检测TAP1对肿瘤免疫应答的作用。采用方差分析做多组比较、独立样本t检验做两两比较。结果 TAP1过表达和敲低3种细胞系成功,细胞计数试剂盒(CCK-8)吸光度中G261细胞的增殖每组无变化和侵袭功能通过t检验,差异无统计学意义(t=1.265,df=4,P>0.05);TAP1的过表达和胶质瘤细胞的敲低表明TAP1和MHC-Ⅰ表达t检验,差异有统计学意义(t=4.560,df=4,P<0.05);TAP1过表达和敲低G261细胞与体内存活和CD8+的免疫化学染色相关(t=5.806,df=4,P<0.05);MHC-Ⅰ类在TAP1过表达和敲低GL261细胞表面表达。结论 TAP1可能通过调节CD8+T细胞而成为胶质瘤的关键抑癌基因。Objective To explore the transporter 1(TAP1)associated with antigen processing is necessary for the major histocompatibility complex-Ⅰ(MHC-Ⅰ)antigen presentation pathway and is an important landmark in tumor immunology monitoring.Methods We overexpressed and knocked down the expression level of TAP1 in three glioma cell lines(U87,U251 and GL261)by lentivirus.The experiment was divided into control group,lentivirus overexpression empty vector control group,lentiviral knockout empty vector control group,TAP1 overexpression group and knockout group.Western blotting and quantitative polymerase chain reactio(qPCR)were used to detect the expression of TAP1 and MHC-Ⅰ,and their interrelationship.Transwell system was used to detect cell proliferation and invasion function.The flow cytometry was used to detect the expression of MHC-Ⅰon the GL261 cell surface with overexpression or knockdown of TAP1.Immunohistochemistry was used to detect the effect of TAP1 on tumor immune response.Results TAP1 overexpression and knockdown were successful in the three cell lines.The proliferation of G261 cells in CCK8 absorbance was unchanged in each group and the invasive function passed the t test,with no statistical difference(t=1.265,df=4,P>0.05);TAP1 overexpression and glioma cell knockdown It showed that TAP1 and MHC-Ⅰexpression had significant difference with(t=4.560,df=4,P<0.01);TAP1 overexpression and knockdown of G261 cells were related to in vivo survival and immunochemical staining of CD8+(t=5.806,df=4,P<0.01);MHC-Ⅰoverexpression and knockdown of GL261 cell surface expression.Conclusion TAP1 may become a key tumor suppressor gene of gliomas by regulating CD8+T cells.

关 键 词：抗原加工相关的转运蛋白1 主要组织相容性复合物-I 抑癌基因 胶质瘤 

分 类 号：R739.41[医药卫生—肿瘤]