应用人类全基因组芯片检测多药耐药白血病细胞株HL-60/VCR与敏感细胞株HL-60基因的表达差异  

Difference of Gene Expression Profiles between HL-60/VCR and HL-60 Cells Detected by Human Genome Genechip

作　　者：董宝侠[1] 陈协群[1] 王哲[2] 梁蓉[1] 白庆咸[1] 黄高昇[2] 张伟平[2] 高广勋[1] 韩冬梅[1] 

机构地区：[1]第四军医大学西京医院血液内科 [2]第四军医大学病理学教研室,西安710032

出　　处：《中国实验血液学杂志》2006年第3期492-496,共5页Journal of Experimental Hematology

基　　金：陕西省自然科学基金资助;编号:2004C-260

摘　　要：本研究利用基因芯片技术检测白血病多药耐药细胞株HL-60/VCR及其亲本细胞株差异基因表达谱,探讨急性白血病多药耐药机制。本试验提取白血病细胞株HL-60及其多药耐药细胞株HL-60/VCR的mRNA,在反转录及体外转录过程中分别用生物素进行标记,与Affymetrix公司人类全基因组芯片U133A杂交,用Affymetrix专用芯片扫描仪G2500AGeneArrayScanner及MicroarraySuite、MicroDBTMGeneChipLIMS、GeneChipDMT分析软件系统处理杂交信号获取结果。结果表明:白血病细胞株HL-60及其耐药株HL-60/VCR差异显示基因5507条,其中耐药细胞株中上调表达基因3100条,下调表达基因2407条;差异极显著性基因1040条,表达上调435条,下调605条,涉及与细胞生长活动及信号转导相关的基因。结论:多基因参与白血病细胞株的多药耐药机制,基因芯片技术是并行分析多个基因、探讨白血病多药耐药机制的有效方法。This study was aimed to detect the gene expression profile changes between human acute leukemia cell line HL-60 and VCR-resistance HL-60, and to investigate the underlying mechanisms of MDR by using genechip technology. In experiments, mRNA were harvested using TrizoL reagent from these two cell lines, through RT-PCR, the biotinylated nucleotide were incorporated into the cRNA during the in vitro transcription reaction. The high quality RNA was hybridized to the gene expression array human genome U133A developed by Affymetrix. It was scanned by G2500A GeneArray Scanner and the acquired image was analysed by a series of softwares. The results showed that 5 507 genes were differentially expressed between human acute leukemia cell line HL-60 and VCR-resistant HL-60. Compared with HL-60, 3 100 genes were up-regulated and 2 407 genes were down-regulated in VCR-resistant cell line. These genes were involved in different cell activities such as growth regulation and signal transduction. Among the genes with remarkable differential expression between the two cell lines, 435 were up-regulated and 605 were down-regulated. It is concluded that many different kinds of genes are involved in the mechanism of MDR and there is an intricate molecular network that controls the sensitivity of leukemia cells to the chemotherapeutic agents. Genechip is an efficient tool for parallel gene expression analysis.

关 键 词：基因芯片 多药耐药 白血病 HL-60 HL-60/VCR 

分 类 号：R733.7[医药卫生—肿瘤]