摘要
目的探讨谷氨酰胺酶2(GLS2)对肝胆管癌细胞5-氟尿嘧啶(5-Fu)化疗敏感性的作用及其机制。方法 TIMER数据库检索GLS2在泛癌中表达水平,通过蛋白质印迹法(Western blot)、免疫组织化学检测进一步验证GLS2在胆管癌组织(2014年1月至2019年4月扬州大学附属医院苏北人民医院行胆管癌根治术患者)表达水平及临床意义。分析GLS2和P-糖蛋白(MDR1)在胆管细胞癌组织中表达相关性。运用pcDNA过表达RBE细胞株GLS2表达;细胞计数试剂盒(CCK-8)法检测各组细胞对化疗药物5-Fu的化疗敏感性。组间比较采用t检验,临床资料采用χ^(2)检验。结果在肝胆管细胞癌组织中GLS2表达显著高于正常组织(0.42±1.20比1.71±0.83,t=2.974,P<0.05),并且GLS2表达与肝胆管细胞癌患者肿瘤数量、有无包膜、淋巴结转移、临床分期、糖类抗原19-9明显有关(χ^(2)=6.17、5.84、7.15、8.24、8.21,P<0.05)。进一步分析表明GLS2表达与MDR1表达明显负相关(r=-0.536,P<0.05)。过表达RBE细胞GLS2表达可下调MDR1表达(0.90±0.12比0.22±0.21,t=3.213,P<0.05),并提高RBE细胞对5-Fu化疗敏感性。结论 GLS2通过介导MDR1表达提高肝胆管癌细胞化疗敏感性,并且GLS2的表达与肝胆管细胞癌患者恶性表型显著相关。
Objective To study the functions and mechanisms of Glutamine2(GLS2)in hepatic cholangiocarcinoma(CCA)cell chemosensitivity to 5-fluorouraeil(5-Fu).Methods Firstly,the expression of GLS2 in Pan cancer was retrieved from Tumor Immune Estimation Resource databases(TIMER)database.Western blotting and immunohistochemistry were used to further verify the expression and clinical significance of GLS2 inCCA(From Janury,2014 to April,2019 patients with CCA underwent radical operation in SUBEI people′s Hospital,Affiliated Hospital of Yangzhou University).Then,the correlation was also analyzed between GLS2 and P-glycoprotein(MDR1).Finally,the regulation of GLS2 in CCA cell were evaluated by pcDNA overexpression.Lastly,we detected the chemosensitivity to 5-Fu by cell counting kit-8(CCK-8)and measured the protein level of MDR1.T test was used for comparison between groups,and the clinical data were analyzed by Chi square test.Results The expression of GLS2 in CCA tissues was significantly up-regulated(0.42±1.20 vs.1.71±0.83,t=2.974,P<0.05),and the expression of GLS2 was significantly correlated with the number of tumors,presence or absence of capsule,lymph node metastasis,clinical stage and CA-199 of CCA patients(χ^(2)=6.17,5.84,7.15,8.24,8.21,P<0.05).Furthermore,the correlation analysis of 40 patients with CCA showed that the expression of GLS2 was negatively correlated with the expression of MDR1(r=-0.536,P<0.05).Overexpression of GLS2 in RBE cells can down regulate the expression of MDR1(0.90±0.12 vs.0.22±0.21,t=3.213,P<0.05)and enhanced chemosensitivity of RBE cells to 5-Fu.Conclusion GLS2 regulated the chemosensitivity of CCA cells to 5-Fu by mediating the expression of MDR1,and the expression of GLS2 is significantly correlated with the malignant phenotype of CCA patients.
作者
曹俊
刘康俊
张弛
蒋国庆
金圣杰
王谦
王翱卿
柏斗胜
Cao Jun;Liu Kangjun;Zhang Chi;Jiang Guoqing;Jin Shengjie;Wang Qian;Wang Aoqing;Bai Dousheng(Medical College of Yangzhou University,Yangzhou 225001,China;Clinical Medicine College of Yangzhou University,Subei People’s Hospital Hepatobiliary Surgery,Institute of General Surgery,Yangzhou 225001,China)
出处
《中华实验外科杂志》
CAS
北大核心
2021年第10期1894-1897,共4页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金(81871909、81702861)。
