硼替佐米耐药多发性骨髓瘤细胞基因表达谱分析  

A microarray analysis of bortezomib-resistant gene expression in multiple myeloma

作　　者：刘琼[1] 蒲业迪[2] 代广霞 马家乐[1] 杨建霞[1] 李丽珍[1] 李颢[1] 王鲁群[1] 

机构地区：[1]山东大学齐鲁医院血液科,山东济南250012 [2]天津市第一中心医院血液科,天津300192

出　　处：《山东大学学报（医学版）》2015年第6期33-38,共6页Journal of Shandong University:Health Sciences

基　　金：山东省科技攻关计划(2008GG1002049);中国医师协会临床医学科研专项基金(20100123)

摘　　要：目的应用基因芯片技术分析硼替佐米敏感细胞株KM3与耐药多发性骨髓瘤细胞株KM3/BTZ基因表达的差异性,探讨硼替佐米耐药相关基因及耐药发生机制。方法采用Affymetrix U133 plus 2.0全基因组表达谱芯片,分析KM3与KM3/BTZ之间的基因表达,采用RT-PCR法进一步检测基因表达的差异性,运用分子注释系统MAS3.0软件和已知耐药相关基因进行数据分析。结果 KM3/BTZ与KM3相比,670个基因的表达具有差异性,表达差异10倍以上基因32个,主要涉及转录调控、信号传导通路等生物效应过程分子;HSPB2、ZNF及M S4A家族部分基因在KM3中低表达,在KM3/BTZ中强阳性表达;RT-PCR法检测显示,除JUN基因,其他7个基因(CA12、CYP1B1、EPB41L3、HSPB2、MS4A4A、SDPR、PAWR)与芯片检测表达差异结果相符。结论在KM3与KM3/BTZ筛选中,ZNF、MS4A家族部分成员以及另外30个表达差异10倍以上的基因,均可能与多发性骨髓瘤硼替佐米耐药相关。全基因数据筛选和耐药相关基因具体分析,是探讨多发性骨髓瘤细胞耐药机制的理想方法之一。Objective To investigate genes related to bortezomib-resistance phenotype by analyzing different gene expression patterns with c DNA microarray and explore the mechanism of bortezomib-resistant myeloma. Methods Gene expression profiles of KM3 / BTZ and KM3 cell lines were compared using Affymetrix U133 plus 2. 0 oligonucleotide microarray. Specific differently-expressed genes were chosen for further verification by real-time RT-PCR. Relevant data were analyzed with molecular annotation system MAS3. 0 software and compared with documented resistance genes.Results It was found that the KM3 / BTZ cell line had 670 differently expressed genes as compared to KM3,of which32 genes were increased or decreased by more than 10 folds. These 32 genes were involved in the regulation of transcription and signal transduction. Some genes of ZNF and MS4 A family and HSPB2 were lowly expressed in KM3,but highly expressed in KM3 / BTZ. Except for JUN,7 genes（ CA12,CYP1B1,EPB41L3,HSPB2,MS4A4 A,SDPR,PAWR） were further verified using real-time RT-PCR and results were consistent with the microarray. Conclusion Some members of ZNF and MS4 A family and another 30 genes whose differential expression is more than 10 folds between KM3 and KM3 / BTZ may be related to bortezomib-resistance in multiple myeloma cells. Combination of filteringgenes and detailed analysis of documented resistance genes provides a potential approach to investigate the mechanism of drug resistance in multiple myeloma cells.

关 键 词：多发性骨髓瘤 硼替佐米 耐药 基因芯片 

分 类 号：R733.3[医药卫生—肿瘤]