氯氮平诱导人脑胶质瘤细胞A172神经元样分化并抑制其增殖  

Neuronal Differentiation and Proliferation Inhibition of Human Glioma Cell A172 Induced by Clozapine

作　　者：汤兆奇 陈振辉 曹畅 叶岩荣 沈赟 TANG Zhaoqi;CHEN Zhenhui;CAO Chang(Department of Pharmacy,Zhongshan Hospital(Xiamen),Fudan University,Fujian 361015,China)

机构地区：[1]复旦大学附属中山医院厦门医院药剂科,361015 [2]厦门市恶性肿瘤综合治疗临床医学研究中心,361015 [3]复旦大学附属中山医院药剂科,上海200032

出　　处：《医学研究杂志》2025年第3期126-131,共6页Journal of Medical Research

基　　金：福建省厦门市自然科学基金资助项目(3502Z202372068)。

摘　　要：目的探讨氯氮平对人脑胶质瘤细胞的神经元分化的诱导作用和对增殖的影响。方法使用苏木精-伊红(hematoxylin-eosin,HE)染色观察氯氮平对人脑胶质瘤细胞A172形态的影响;使用免疫细胞化学检测神经元标志物神经元核抗原(neuronal nuclei antigen,NeuN)和微管相关蛋白2(microtubule-associated protein 2,MAP2)的表达,以及星形胶质细胞标志物胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)与S100钙结合蛋白B(S100 calcium binding protein B,S100B)和少突胶质细胞标志物髓鞘碱性蛋白(myelin basic protein,MBP)与转录因子2(Olig2)的表达;使用ki-67法、CCK-8法和克隆形成实验检测氯氮平对A172细胞增殖影响;使用SwissTargetPrediction、GeneCards、STRING数据库预测氯氮平潜在的作用机制;使用Western blot法检测β-catenin的表达水平。结果氯氮平(30μmol/L)使A172细胞形态发生改变,细胞出现细长突起;氯氮平诱导A172细胞表达神经元标志物NeuN和MAP2,而没有星形胶质细胞和少突胶质细胞标志物表达;氯氮平显著抑制A172细胞增殖;基于数据库的分析提示氯氮平作用机制可能与Wnt信号通路有关;氯氮平下调A172细胞β-catenin表达。结论氯氮平诱导人脑胶质瘤细胞A172神经元样分化并抑制其增殖。Objective To investigate the effect of clozapine on neuronal differentiation and proliferation of human glioma cells.Methods Hematoxylin-eosin(HE)staining was used to observe the effect of clozapine on the morphology of human glioma cell A172.Immunocytochemistry was used to detect the expression of neuronal markers neuronal nuclei antigen(NeuN)and microtubule-associated protein 2(MAP2),as well as the expression of astrocytic markers glial fibrillary acidic protein(GFAP)and S100 calcium binding protein B(S100B),and oligodendrocytic markers myelin basic protein(MBP)and Olig2.ki-67detection,CCK-8 assay and clone formation assay were used to assess the effects of clozapine on the cell proliferation of A172.SwissTargetPrediction,GeneCards,and STRING databases were utilized to predict the potential mechanism of clozapine.Western blot was used to detect the expression level ofβ-catenin.Results Clozapine(30μmol/L)changed the morphology of A172 cells,with elongated cell projection.Clozapine induced the expression of neuronal markers NeuN and MAP2 in A172 cells,without inducing the expression of astrocytic or oligodendrocytic markers.Clozapine significantly inhibited the proliferation of A172 cells.Database analysis suggested that the mechanism of clozapine might be related to the Wnt signaling pathway.Clozapine down-regulated the expression ofβ-catenin in A172 cells.Conclusion Clozapine induced neuronal differentiation of human glioma cells A172,and inhibited their proliferation.

关 键 词：氯氮平 胶质瘤 分化 神经元 增殖 

分 类 号：R971[医药卫生—药品]