胶质瘤SLC22A18耐药细胞株的建立及其耐药机制的研究  

Establishment of SLC22A18 drug-resistant gene in glioma cells and investigation of its drug resistance mechanism

作　　者：杨标 楚胜华[1] 马延斌[1] Yang Biao;Chu Shenghua;Ma Yanbin(Departmet of Neurosurgery,Shanghai Ninth People's Hospital Affiliated to Shanghai Jiaotong University School of Medicine,Shanghai 201999,China)

机构地区：[1]上海交通大学医学院附属第九人民医院神经外科,上海201999

出　　处：《现代肿瘤医学》2019年第5期719-722,共4页Journal of Modern Oncology

基　　金：教育部留学回国人员科研启动基金项目(编号:082003);上海市卫生和计划生育委员会科研基金面上项目(编号:201540266);上海交通大学医工交叉研究基金项目(编号:YG2015MS25);上海交通大学医学院附属第三人民医院自然科学研究基金项目(编号:syz2015-015)

摘　　要：目的:以人胶质瘤U251细胞株为载体,探讨胶质瘤获得性SLC22A18基因耐药的可能机制。方法:用重组腺病毒介导的SLC22A18基因(Ad/SLC22A18)反复处理人胶质瘤U251细胞,得到人胶质瘤耐药细胞株U251-SLC22A18/R。通过MTT和Western blot法检测耐药细胞U251-SLC22A18/R对Ad/SLC22A18杀伤作用的敏感性及凋亡相关的信号分子DR4、DR5、Bax、Bcl-XL和caspase-8的表达情况,分析其获得性耐药的可能机制。结果:耐药细胞U251-SLC22A18/R对Ad/SLC22A18和SLC22A18蛋白处理耐药,但对Bax基因处理仍然敏感。U251-SLC22A18/R细胞内Bcl-XL的表达显著升高,caspase-8的表达明显下降,caspase-8活性未见明显的裂解。结论:人胶质瘤U251细胞株经Ad/SLC22A18反复处理后产生针对SLC22A18基因的耐药,其机制可能与Bcl-XL表达升高和caspase-8表达下降有关。Objective:To investigate the drug resistance mechanism involved in acquiring the SLC22A18 drug-resistant gene in huma glioma U251 cell line.Methods:Human glioma U251 cells were repeatedly treated with adenovirus-mediated SLC22A18 gene(Ad/SLC22A18),and U251-SLC22A18/R cells resistant to Ad/SLC22A18 were obtained.The cell growth was analyzed using MTT method.The expression of some proteins involved in apoptosis pathway was measured by Western blot analysis.Then the mechanism involved in acquiring drug resistance was analyzed.Results:U251-SLC22A18/R cells resistant to SLC22A18 were obtained from SLC22A18-sensitive U251 parent cells.But they were still sensitive to adenovirus-mediated Bax gene therapy.Further studies demonstrated that expression of Bcl-XL was significantly upregulated whereas caspase-8 was dramatically downregulated in U251-SLC22A18/R cells,and there was no cleaved caspase-8 found in the U251-SLC22A18/R cells.Conclusion:After Ad/SLC22A18 repeated treatment,U251 cells could acquire the resistance to SLC22A18 gene.The possible mechanism may involve upregulation of Bcl-XL and downregulation of caspase-8.

关 键 词：SLC22A18 基因治疗 胶质瘤 耐药 

分 类 号：R730.264[医药卫生—肿瘤]