神经胶质瘤C6/VP16多药耐药细胞株的建立  

Establishment of multidrug resistant glioma cell line C6/VP16

作　　者：李乾锋[1] 徐海涛[1] 王龙[1] 黄书岚[1] 

机构地区：[1]武汉大学人民医院神经外科,武汉430060

出　　处：《中国临床神经外科杂志》2012年第7期414-416,420,共4页Chinese Journal of Clinical Neurosurgery

摘　　要：目的建立神经胶质瘤C6细胞对依托泊苷(VP16)耐药细胞株C6/VP16,探讨其耐药的可能机制。方法采用浓度递增和短期大剂量药物诱导相结合的方法建立大鼠C6/VP16耐药细胞株;光镜下观察C6/VP16细胞株的形态学变化;细胞计数试剂盒-8检测C6/VP16细胞株对VP16、阿霉素(ADM)、长春新碱(VCR)、环磷酰胺(CTX)、替莫唑胺(TMZ)的耐药敏感性;westernblot检测多药耐药相关蛋白1(MRP1)的表达量变化。结果经过65代诱导培养,成功建立可操作的、重复性好的C6/VP16耐药细胞株。C6亲代细胞形态规则、大小均一,生长迅速,紧密生长,每2～3d传代;C6/VP16细胞株,突起增多,生长周期受抑制,生长变缓,每10～12d传代。VP16、TMZ、CTX、ADM和VCR作用C6亲代细胞的半生长抑制浓度(IC50)分别为(0.105±0.043)、(27.284±0.529)、(5.800±0.317)、(6.636±0.315)和(19.146±0.526)μg/ml,而作用C6/VP16细胞株的IC50分别为(0.943±0.052)、(33.251±0.288)、(32.943±0.215)、(35.251±0.126)和(35.604±0.426)μg/ml;VP16、CTX和ADM作用C6/VP16细胞株IC50较C6亲代细胞差异显著(P<0.05)。C6/VP16细胞株MRP1的表达量较亲代C6细胞明显增加(P<0.05)。结论 C6/VP16细胞株对ADM和CTX存在明显交叉耐药,而对TMZ和VCR无明显交叉耐药;多药耐药性是多种机制相互作用的结果,MRP1可能参与其中。Objective To establish a multidrug resistant（MDR）glioma cell line in order to study the mechanism of MDR.M ethods An etoposide（VP16）resistant glioma cell line C6/VP16 was established by exposure of C6 cells to the gradient concentration and a big dose VP16.The changes in C6/VP16 morphology were observed under a light-microscope.The sensitivity of C6/VP16 to VP16,adriamycin（ADM）,vincristine（VCR）,cytoxan（CTX）and temozolomide（TMZ）were determined by cell counting kit-8（CCK-8）.The expression of multidrug resistance associated protein 1（MRP1）was detected by western blot.Results VP16 resistant glioma cell line C6/VP16 was successfully established after passaging 65 generations.The half growth inhibition concentration（IC 50）of VP16,TMZ,C TX,ADM and VCR to C6 cells were（0.105±0.043）,（27.284±0.529）,（5.800±0.317）,（6.636±0.315）and（19.146±0.526）μg/ml respectively.IC 50 of VP16,TMZ,CTX,ADM and VCR to C6/VP16 cells were（0.943±0.052）,（33.251±0.288）,（32.943±0.215）,（35.251±0.126）and（35.604±0.426）μg/ml respectively.IC 50 of VP16,CTX and ADM to C6/VP16 cells were significantly higher than those to C6 cells（P〈0.05）.The level of MRP1 expression（3.97±2.10）in C6/VP16 was significantly higher than that（1.84±1.11）in C6 cells（P〈0.05）.Conclusions C6/VP16 is of a multidrug and stable resistance.The increase in MRP1 expression may be one of the mechanisms of C6/Vp16 MDR.

关 键 词：神经胶质瘤 C6细胞 多药耐药性 多药耐药相关蛋白1 依托泊苷 

分 类 号：R73-351[医药卫生—肿瘤] R730.53[医药卫生—临床医学]