CAJ | 학술논문

目的研究人脑胶质瘤细胞系的耐药与鞘糖脂的相关性.方法采用MTT方法检测了人脑胶质瘤细胞系A172和U251对替尼泊苷的药物敏感性,然后质谱分析两株细胞系的鞘糖脂水平的变化,并采用逐渐增加药物浓度法建立U251细胞系耐替尼泊苷细胞系(U251/VM-26细胞系),然后MTT法检测其对替尼泊苷的敏感度、Real-time PCR技术检测其耐药基因MDR1和MRP1的表达、质谱分析细胞系耐药前后鞘糖脂的变化.结果 MTT显示A172细胞对替尼泊苷的耐药性高于U251细胞,同时质谱分析发现A172细胞中C18-LacCer、C18-Gb3、C18-Gb4鞘糖脂表达明显高于U251细胞系.U251细胞系经诱导耐药后(U251/VM-26细胞系)其耐药性上升100倍(P＜0.001)；同时发现U251细胞系药物诱导耐药后C18-Gb4的表达上升,C18-LacCer和C18-Gb3鞘糖脂表达也上升.Real-time PCR显示A172细胞系中MRP1基因表达水平显著高于U251细胞系,U251/VM-26耐药细胞系中MRP1、MDR1基因显著高于U251细胞系.结论 C18-LacCer、C18-Gb3和C18-Gb4鞘糖脂与A172、U251和U251/VM-26胶质瘤细胞系对替尼泊苷的敏感性相关,且耐药基因MDR1和MRP1参与了人脑胶质瘤细胞系对替尼泊苷的耐药.
목적 연구인뇌효질류세포계적내약여초당지적상관성.방법 채용MTT방법검측료인뇌효질류세포계A172화U251대체니박감적약물민감성,연후질보분석량주세포계적초당지수평적변화,병채용축점증가약물농도법건립U251세포계내체니박감세포계(U251/VM-26세포계),연후MTT법검측기대체니박감적민감도、Real-time PCR기술검측기내약기인MDR1화MRP1적표체、질보분석세포계내약전후초당지적변화.결과 MTT현시A172세포대체니박감적내약성고우U251세포,동시질보분석발현A172세포중C18-LacCer、C18-Gb3、C18-Gb4초당지표체명현고우U251세포계.U251세포계경유도내약후(U251/VM-26세포계)기내약성상승100배(P＜0.001)；동시발현U251세포계약물유도내약후C18-Gb4적표체상승,C18-LacCer화C18-Gb3초당지표체야상승.Real-time PCR현시A172세포계중MRP1기인표체수평현저고우U251세포계,U251/VM-26내약세포계중MRP1、MDR1기인현저고우U251세포계.결론 C18-LacCer、C18-Gb3화C18-Gb4초당지여A172、U251화U251/VM-26효질류세포계대체니박감적민감성상관,차내약기인MDR1화MRP1삼여료인뇌효질류세포계대체니박감적내약.
Objective To study the relationship of the drug resistance and glycosphingolipids in glioma.Methods The sensitivity of the teniposide in A172 and U251 cells were detected by MTT,and their glycosphingolipids were detected by ESI-LIT-MS.The drug resistance cell line U251/VM-26 was established by culture with gradual increased dose of VM-26.Then the sensitivity of the teniposide in U251 and U251/VM-26 were checked by MTT,the drug resistance associated gene MDR1 and MRP1 were detected by real-time PCR,and their glycosphingolipids were detected by ESI-LIT-MS.Results The drug resistant of A172 were higher than U251 cells.Meanwhile,C18-LacCer,C18-Gb3,C18-Gb4,are highly accumulated in A172 cells than in U251 cells.The drug resistant of U251/VM-26 cells was 100-folds higher than U251 cells after co-cuturing with VM-26,then the levels of C18-Gb4 are significantly increased in U251/VM-26 cells,C18-LacCer and C18-Gb3 are also increased.Real-time PCR data show that MRP1 in A172 cells are higher than U251 cells,the level of MDR1 and MRP1 gene are also highly accumulated in U251/VM-26 cells.Conclusions Glioma-associated glycosphingolipids,C18-LacCer、C18-Gb3 and C18-Gb4,are correlated with drug resistance of VM-26,and MDR1 and MRP1 gene are involved in the drug resistant of those glioma ceils.