β1,3-N-乙酰氨基葡萄糖转移酶5靶向调控膜联蛋白A2促进神经母细胞瘤细胞侵袭迁移  

β1,3-N-acetylglucosaminyltransferase-V promotes the invasion and migration of neuroblastoma cells by regulating Annexin A2

作　　者：向田 蔡秦真 刘杲 刘鑫袁 纯辉 Xiang Tian;Cai Qinzhen;Liu Gao;Liu Xin;Yuan Chunhui(Hubei Selenium and Human Health Institute,Department of Laboratory Medicine,Central Hospital of Enshi Tujia and Miao Autonomous Prefecture,Enshi 445000,China;Department of Laboratory Medicine,Wuhan Children’s Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430016,China;Department of Gastrointestinal Surgery,Central Hospital of Enshi Tujia and Miao Autonomous Prefecture,Enshi 445000,China;Department of Pediatric Surgery,Wuhan Children’s Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430016,China)

机构地区：[1]恩施土家族苗族自治州中心医院检验科、湖北硒与人体健康研究院,恩施445000 [2]华中科技大学同济医学院附属武汉儿童医院检验科,武汉430016 [3]恩施土家族苗族自治州中心医院胃肠外科,恩施445000 [4]华中科技大学同济医学院附属武汉儿童医院普外科,武汉430016

出　　处：《中华实验外科杂志》2023年第2期295-297,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(82060539、81760540);武汉市卫生与健康委员会自然科学基金(WX18Q03)。

摘　　要：目的探讨β1,3-N-乙酰氨基葡萄糖转移酶5(B3GNT5)对神经母细胞瘤(NB)细胞侵袭迁移的影响及作用机制。方法利用公共数据库分析B3GNT5与NB患者高/低风险、MYCN扩增及预后的关系。收集武汉儿童医院普外科7例NB和2例节神经细胞瘤组织标本,采用免疫组织化学、实时定量荧光聚合酶链反应(RT-qPCR)和蛋白印迹法(Western blot)检测组织和NB细胞系中B3GNT5的表达。构建B3GNT5干扰和过表达的SK-N-BE和SH-SY-5Y细胞系,Transwell检测对细胞迁移侵袭的影响,Western blot检测转移相关蛋白。应用质谱技术、免疫共沉淀、靶基因干扰实验探讨潜在分子机制。两组间比较采用t检验。结果数据库分析显示B3GNT5表达与NB患儿总体生存率和无事件生存率呈负相关[风险比(HR)=13.480、3.900,P<0.01];高危组和MYCN扩增组中的表达显著高于低危组和MYCN非扩增组(10.040±1.206比9.206±0.893、10.960±0.705比9.166±0.933,t=8.816、14.900,P<0.01)。B3GNT5干扰组SK-N-BE细胞迁移(78.980±6.591)和侵袭(39.830±7.310)的数量低于对照组(131.000±6.607、72.000±6.117,t=23.720、14.320,P<0.01)。B3GNT5与膜联蛋白A2结合,B3GNT5干扰组膜联蛋白A2糖基化水平低于对照组(0.217±0.076比0.470±0.071,t=4.192,P<0.05)。干扰膜联蛋白A2,B3GNT5过表达的SK-N-BE细胞迁移(91.780±6.873)和侵袭(99.110±7.435)的数量低于对照组(188.900±7.727、197.200±8.363,t=39.860、36.730,P<0.01)。结论B3GNT5靶向促进膜联蛋白A2糖基化修饰介导NB细胞侵袭迁移。Objective To investigate the effect of B3GNT5 on the metastasis of neuroblastoma(NB)and its mechanism.Methods The public database was used to evaluate the correlation between B3GNT5 and prognosis in NB and the expression of B3GNT5 in different risk and MYCN status.A total of 7 cases of NB and 2 cases of ganglioneuroma tissues were collected from the Department of Pediatric Surgery of Wuhan Children’s Hospital.Immunohistochemistry,quantitative real-time polymerase chain reaction(RT-qPCR)and Western blotting were used to detect the expression of B3GNT5 in clinical tissues and different NB cell lines.B3GNT5 was interfered/overexpressed in SK-N-BE and SH-SY-5Y cells,migration and invasion ability was detected by Transwell assay,and metastasis related proteins were detected by Western blotting.The molecular mechanism was investigated by mass spectrometry,immunoprecipitation and target gene interference.Results B3GNT5 expression was significantly higher in high-risk and MYCN amplified group(10.040±1.206 vs.9.206±0.893,10.960±0.705 vs.9.166±0.933,t=8.816,14.900,P<0.01),and predicted poor overall survival and event-free survival of NB patients[hazard ratio(HR)=13.480,3.900,P<0.01].Interfering B3GNT5 inhibited the migration(78.980±6.591 vs.131.000±6.607)and invasion(39.830±7.310 vs.72.000±6.117,t=23.720,14.320,P<0.01)of SK-N-BE cells.B3GNT5 could bind to Annexin A2.Interfering B3GNT5 significantly down-regulated the glycosylation of Annexin A2(0.217±0.076 vs.0.470±0.071,t=4.192,P<0.05).Interfering Annexin A2 blocked the promoting effects of B3GNT5 on migration(91.780±6.873 vs.188.900±7.727)and invasion(99.110±7.435 vs.197.200±8.363,t=39.860,36.730,P<0.01)of SK-N-BE cells.Conclusion B3GNT5 can upregulate Annexin A2 glycosylation to promote the invasion and migration of NB cells.

关 键 词：神经母细胞瘤 膜联蛋白A2 糖基化 侵袭 迁移 

分 类 号：R739.4[医药卫生—肿瘤]