天冬酰胺合成酶通过促进β-catenin核转位驱动胆管癌转移  

Asparagine synthetase promotes cholangiocarcinoma metastasis by facili⁃tating nuclear translocation ofβ-catenin

作　　者：褚珍珍 周栩萱 刘力豪 张鲍欢 姚楠 CHU Zhenzhen;ZHOU Xuxuan;LIU Lihao;ZHANG Baohuan;YAO Nan(Department of Pathophysiology,School of Medicine,Jinan University,Guangzhou 510632,China;Laboratory of Patho-physiology,National Administration of Traditional Chinese Medicine,Jinan University,Guangzhou 510632,China;Morpholo-gy Experimental Teaching Center,School of Medicine,Jinan University,Guangzhou 510632,China)

机构地区：[1]暨南大学基础医学院病理生理学系,广东广州510632 [2]国家中医药管理局病理生理科研实验室,广东广州510632 [3]暨南大学基础医学院形态学实验教学中心,广东广州510632

出　　处：《中国病理生理杂志》2023年第9期1537-1546,共10页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82172602);广东省自然科学基金资助项目(No.2023A1515011892)。

摘　　要：目的:检测天冬酰胺合成酶(ASNS)在胆管癌(CCA)中的表达情况,探讨ASNS在CCA转移中的作用及其机制。方法:通过公共数据库分析各肿瘤组织中ASNS的mRNA表达;收集CCA患者病理组织(n=27),构建硫代乙酰胺诱导的大鼠自发CCA模型和左中位胆管结扎联合二乙基亚硝胺诱导的小鼠自发CCA模型,通过免疫组化、Western blot和免疫荧光法检测ASNS蛋白表达。采用CCK8、划痕和Transwell实验检测ASNS对人CCA细胞HuCCT1和HCCC-9810增殖、迁移和侵袭的影响。构建ASNS稳定敲减的CCA细胞株HuCCT1^(shNC)、HuCCT1^(shASNS)、HCCC-9810^(shNC)和HCCC-9810^(shASNS),通过肝原位种植和尾静脉注射研究ASNS对CCA细胞肝内生长和肺转移的影响。利用公共数据库富集与ASNS相关的信号通路,并用免疫荧光和Western blot验证相关分子机制。结果:无论在人或动物CCA组织中,ASNS表达水平均高于癌旁组织(P<0.01)。ASNS以酶活性非依赖性方式促进CCA细胞HuCCT1和HCCC-9810的增殖、迁移与侵袭。生物信息学分析显示,β-catenin在ASNS高表达的CCA组织中富集,ASNS通过促进β-catenin核转位,启动CCA细胞上皮-间充质转化(EMT)。β-catenin抑制剂XAV-939可显著抑制CCA细胞的侵袭与迁移。结论:ASNS在CCA中高表达,通过促进β-catenin核转位,介导EMT,驱动CCA转移。AIM:To investigate the expression of asparagine synthetase(ASNS)in cholangiocarcinoma(CCA),and to explore the potential role and molecular mechanisms of ASNS in CCA metastasis.METHODS:A public database was used to analyze ASNS mRNA expression in various tumor tissues.Immunohistochemistry,Western blot and immunofluorescence were used to detect ASNS protein expression in human CCA specimens and murine spontaneous CCA models.The CCK8,wound-healing and Transwell assays were performed to examine the effects of ASNS on CCA cell pro-liferation,migration and invasion.The effects of ASNS on CCA intrahepatic growth and lung metastasis were investigated by orthotopic implantation and tail vein injection of stable ASNS knockdown CCA cell lines.The ASNS-related pathways in CCA were examined using gene set enrichment analysis,and the underlying molecular mechanisms were verified by immu-nofluorescence and Western blot.RESULTS:The expression of ASNS in both human and murine CCA tissues was higher than that in adjacent noncancerous tissues.ASNS promoted the proliferation,migration and invasion of human CCA HuCCT1 and HCCC-9810 cells in an enzyme activity-independent manner.Bioinformatics analysis showed thatβ-catenin was enriched in CCA tissues with high expression of ASNS,and ASNS initiated epithelial-mesenchymal transition(EMT)of CCA by promoting the nuclear translocation ofβ-catenin.Blocking the nuclear translocation ofβ-catenin with XAV-939 significantly inhibited the invasion and migration of CCA cells.CONCLUSION:ASNS is highly expressed in CCA and promotes EMT by meditating the nuclear translocation ofβ-catenin.

关 键 词：胆管癌 肿瘤转移 天冬酰胺合成酶 Β-CATENIN信号通路 上皮-间充质转化 

分 类 号：R575.7[医药卫生—消化系统] R363[医药卫生—内科学]