MGMT反义RNA对人脑胶质瘤耐药性逆转的研究  

Reveral of drug resistance of human glioma by MGMT antisense RNA

作　　者：金澎[1] 张庆林[1] 刘福生[2] 王忠诚[2] 魏林[1] 王成伟[1] 孔建新[1] 

机构地区：[1]山东大学第二医院神经外科,济南250033 [2]北京市神经外科研究所

出　　处：《中华神经外科杂志》2003年第2期93-98,共6页Chinese Journal of Neurosurgery

基　　金：国家自然科学基金资助项目 ( 30 0 70 2 72 )

摘　　要：目的　利用反义技术逆转胶质瘤的耐药性。方法　模拟临床亚硝基脲类药物的用药程序 ,在体外建立由DNA修复蛋白O6 甲基鸟嘌呤 DNA甲基转移酶 (MGMT)介导的人脑胶质瘤耐药细胞系U2 51 /BCNU ,并进行细胞耐药性检测和细胞MGMTmRNA表达的分析 ;观察MGMT反义RNA对U2 51 /BCNU细胞MGMT表达的调节作用及对U2 51 /BCNU细胞BCNU敏感性的影响。结果　经过反复总共 5次的用药过程 ,首次在体外建立了对BCNU具有稳定抗性的U2 51 /BCNU ,其对BCNU的耐受程度约为U2 51的 1 7倍 ;RT PCR结果显示 ,U2 51 /BCNU细胞有MGMTmRNA的表达。MGMT反义RNA表达载体pLaMT5SN和pLaMTSN能够在一定程度上降低U2 51 /BCNU细胞中MGMTmRNA的表达水平 ,提高其对BCNU的敏感性。结论　MGMT反义RNA能够在一定程度上抑制MGMT的表达水平 。Objective To reverse the drug resistance of glioma by antisense RNA.Methods Simulating the clinical way of nitrosoureas, to establish a BCNU resistant human glioma subline U251/BCNU. To detect the resistance factor and the expression of MGMT mRNA of U251/B CNU. To observe the effect of MGMT antisense RNA on MGMTmRNA expression by RT PCR and the sensitivity to BCNU by MTT assay of U251/BCNU. Results First established in vitro a BCNU resistant human glioma subline U251/BCNU through five repeating medications. U251/ BCNU cells showed 17 fold higher resistance to BCNU than did U251 cells by MTT assay. RT PCR suggested that pLaMT5SN and pLaMTSN could down regulated the expression of MGMTmRNA. The sensitivity of aMT5U and aMTU cells to BCNU were 7 and 2 fold higher than that of U251/BCNU cells by MTT assay, which suggested that pLaMT5SN and pLaMTSN could enhance the sensitivity of U251/BCNU to BCNU. Conclusions MGMT antisense RNA could down regulate the expression of MGMT, and enhance the sensitivity of tumor cells to nitrosoureas. Antisense RNA is an effective way to reverse the drug resistance of gliomas, and has prospects for clinical application in the future.

关 键 词：脑胶质瘤 耐药性 O^6-甲基鸟嘌呤-DNA甲基转移酶 反义RNA 逆转作用 

分 类 号：R739.4[医药卫生—肿瘤]