胶质母细胞瘤中癌-睾丸抗原OY-TES-1的表达对M2型巨噬细胞极化的影响  

Effect of cancer-testis antigen OY-TES-1 expression on the polarization of M2 macrophages in glioblastoma

作　　者：赵振凯 梁国 李枫 农蔚霞[1,2] 张庆梅 罗彬[1] 谢小薰 ZHAO Zhenkai;LIANG Guo;LI Feng;NONG Weixia;ZHANG Qingmei;LUO Bin;XIE Xiaoxun(Teaching and Research Section of Histology and Embryology,Guangxi Medical University,Nanning 530021,China;Key Labo-ratory of Basic Research on Regional Diseases in Guangxi Colleges and Universities,Nanning 530021,China)

机构地区：[1]广西医科大学组织学与胚胎学教研室,南宁530021 [2]广西高校区域性疾病基础研究重点实验室,南宁530021

出　　处：《中国临床新医学》2024年第8期875-880,共6页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

基　　金：国家自然科学基金地区科学基金项目(编号:81960453);广西自然科学基金项目(编号:2023GXNSFAA026120)。

摘　　要：目的探讨胶质母细胞瘤(GBM)中癌-睾丸抗原OY-TES-1的表达对M2型巨噬细胞极化的影响。方法体外培养人GBM细胞U251与人外周血单核细胞THP-1,构建稳定下调OY-TES-1的U251稳转株(U251-SH-OY-TES-1组),将转染无关序列的U251作为对照(U251-SH-NC组)。48h后取两组上清液与THP-1细胞共培养,加入U251-SH-NC组上清液的THP-1细胞为SH-NC组,加入U251-SH-OY-TES-1组上清液的THP-1细胞为SH-OY-TES-1组。通过实时荧光定量聚合酶链反应(RT-qPCR)和蛋白免疫印迹法分别检测两组OY-TES-1mRNA和蛋白相对表达量,确定转染效率。通过酶联免疫吸附试验(ELISA)检测培养液中人白细胞介素-10(IL-10)、人转化生长因子-β(TGF-β)表达水平。通过流式细胞术检测CD206表达水平。结果RT-qPCR与蛋白免疫印迹法检测结果显示,U251-SH-OY-TES-1组OY-TES-1mRNA和蛋白的相对表达量低于U251-SH-NC组,差异有统计学意义(P<0.05)。ELISA结果显示,两组培养液中TGF-β、IL-10水平随培养时间的增加呈上升趋势。第0天、第2天SH-OY-TES-1组培养液中TGF-β水平显著低于SH-NC组(P<0.05),IL-10水平比较差异无统计学意义(P>0.05)。第4天SH-OY-TES-1组培养液中TGF-β、IL-10水平显著低于SH-NC组(P<0.05)。流式细胞术结果显示,与SH-NC组比较,SH-OY-TES-1组CD206水平下降,差异有统计学意义(P<0.05)。结论GBM中OY-TES-1高表达能够促进M2型巨噬细胞的极化。Objective To explore the effect of cancer-testis antigen OY-TES-1 expression on the polarization of M2 macrophages in glioblastoma(GBM).Methods Human GBM cell U251 and human peripheral blood mononuclear cell THP-1 were cultured in vitro to construct a stable transfection cell line U251 that stably down-regulated OY-TES-1(U251-SH-OY-TES-1 group),and U251 cells transfected with unrelated sequences were used as the controls(U251-SH-NC group).Forty-eight hours later,the supernatant of the two groups was co-cultured with THP-1 cells.THP-1 cells to which the supernatant of the U251-SH-NC group was added were taken as the SH-NC group,and THP-1 cells to which the supernatant of the U251-SH-OY-TES-1 group was added were taken as the SH-OY-TES-1 group.The relative expres-sion levels of OY-TES-1 mRNA and protein in the two groups were detected by using real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blotting,respectively,and the transfection efficiency was determined.The expression levels of human interleukin-10(IL-10)and human transforming growth factor-β(TGF-β)in the culture solution were detected by enzyme-linked immunosorbent assay(ELISA).The expression level of CD206 was detected by flow cytometry.Results The detection results of RT-qPCR and Western blotting method showed that the relative expressions of OY-TES-1 mRNA and protein in the U251-SH-OY-TES-1 group were lower than those in the U251-SH-NC group,and the differences were statistically significant(P<0.05).The results of ELISA showed that the levels of TGF-βand IL-10 in the culture solution of the two groups increased with the increase of culture time.On day 0 and day 2,the levels of TGF-βin the culture solution of the SH-OY-TES-1 group were significantly lower than those of the SH-NC group(P<0.05),and there were no significant differences in the levels of IL-10 on day 0 and day 2 between the two groups(P>0.05).On day 4,the levels of TGF-βand IL-10 in the culture solution of the SH-OY-TES-1 group were significantly lower tha

关 键 词：胶质母细胞瘤 OY-TES-1 M2型巨噬细胞 极化 

分 类 号：R392[医药卫生—免疫学]