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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 analyze the difference of KM3 / BTZ gene expression between KMT-resistant bortezomib cell line KM3 and multiple myeloma KM3 / BTZ gene by gene chip technique, and to explore the gene and mechanism of resistance to bortezomib. Methods Affymetrix U133 plus 2.0 genome-wide expression microarray was used to analyze the gene expression between KM3 and KM3 / BTZ. RT-PCR was used to detect the difference of gene expression. Using molecular annotation system MAS3.0 software and known drug resistance Related genes for data analysis. Results The expression of 670 genes was different between KM3 / BTZ and KM3. There were 32 genes with more than 10-fold difference between KM3 / BTZ and KM3. These genes mainly involved in transcriptional regulation and signal transduction pathways. Some genes of HSPB2, ZNF and M S4A family In KM3, it was strongly expressed in KM3 / BTZ. The RT-PCR assay showed that the expression levels of the other 7 genes (CA12, CYP1B1, EPB41L3, HSPB2, MS4A4A, SDPR, PAWR) The result is consistent. Conclusion In KM3 and KM3 / BTZ screening, some members of ZNF and MS4A families and another 30 genes with more than 10-fold difference may be associated with bortezomib resistance in multiple myeloma. The whole gene data screening and drug resistance-related gene specific analysis, is to explore the mechanism of multiple myeloma cells is an ideal method of resistance.