胶质瘤干细胞的结肠癌转移相关基因1表达与替莫唑胺耐药的相关性  

Correlation between the expression of MACC1 and drug resistance of temozolomide in glioma stem cells

作　　者：吕华荣[1] 胡胜[1] 吴星[1] 荣亮[1] 申松波[1] 方媛 LV Hua-rong;HU Sheng;Wu Xing(Department of Neurosurgery,Huangshi Central Hospital of Edong Healthcare,Huangshi 435000,China)

机构地区：[1]鄂东医疗集团黄石市中心医院神经外科,黄石435000 [2]华中科技大学同济医学院附属协和医院神经外科

出　　处：《临床神经外科杂志》2019年第6期476-480,共5页Journal of Clinical Neurosurgery

基　　金：湖北省自然科学基金项目(2014CFB421);湖北省黄石市医疗卫生科技攻关计划项目[(2010)1号]

摘　　要：目的探讨结肠癌转移相关基因1(MACC1)表达与胶质瘤干细胞对替莫唑胺(TMZ)耐药的相关性。方法采用实时定量荧光PCR和Western blot方法测定U87胶质瘤细胞来源的干细胞(GSCs-U87)、U251胶质瘤细胞来源的干细胞(GSCs-U251)及U87和U251细胞的MACC1 mRNA、蛋白表达水平。通过实时定量荧光PCR和Western blot检测RNA干扰对MACC1的沉默效应。用流式细胞术和CCK-8方法测定MACC1沉默对TMZ诱导的细胞毒性和细胞凋亡的影响。结果胶质瘤干细胞GSCs-U87和GSCs-U251中MACC1 mRNA和蛋白表达水平显著增高(均P<0.05)。shRNA-MACC1可以显着抑制MACC1表达(均P<0.05)。用不同浓度的TMZ处理后,与对照组GSCs-U87和GSCs-U251相比,TMZ对MACC1沉默的GSCs-U87和GSCs-U251细胞毒性显著增加,且MACC1沉默的胶质瘤干细胞的凋亡数增高。结论MACC1基因的沉默可增强TMZ对胶质瘤细胞的凋亡和生长抑制作用,从而增加其对TMZ化疗的敏感性。Objective To investigate the relationship between metastasis-associated in colon cancer 1(MACC1)gene expression and the resistance of glioma stem cell to temozolomide(TMZ).Methods The expression of MACC1 in U87 glioma cell-derived stem cells(GSCs-U87),U251 glioma cell-derived stem cells(GSCs-U251),U87 and U251 was determined by quantitative real-time PCR and Western blotting.The silencing effect of RNA interference on MACC1 was detected by quantitative real-time fluorescence PCR and Western blot.The effect of MACC1 silencing on TMZ-induced cytotoxicity and apoptosis was measured by flow cytometry and CCK-8.Results The expression of MACC1 in GSCs-U87 and GSCs-U251 in glioma stem cells was up-regulated remarkably at mRNA and protein level(all P<0.05).shRNA-MACC1 significantly inhibited MACC1 gene expression(all P<0.05).The cytotoxicity of TMZ at various concentrations significantly increased in MACC1-silenced GSCs-U87 and GSCs-U251 cells compared with the control group GSCs-U87 and GSCs-U251.In addition,the apoptosis rate of MACC1-silenced GSCs-U87 and GSCs-U251 induced by TMZ significantly increased compared with the control group GSCs-U87 and GSCs-U251.Conclusions There is a relationship between overexpression of MACC1 and the characteristics of glioma stem cells.The silencing of the MACC1 gene enhances the effect of TMZ on the apoptosis and growth of glioma stem cells,thereby increasing the sensitivity of glioma stem cells to TMZ chemotherapy.

关 键 词：胶质细胞瘤 MACC1 替莫唑胺 药物抵抗 

分 类 号：Q344.13[生物学—遗传学]