葡萄糖神经酰胺合酶与白血病细胞耐药的关系  

Relation Between Glucosylceramide Synthase and Multidrug Resistance in Leukemia Cells

作　　者：谢平[1] 葛素梅[2] 沈云峰[3] 顾中华[1] 穆会君[1] 张滨[1] 

机构地区：[1]南京医科大学附属无锡第一医院中心实验室,无锡214002 [2]苏州大学病理生理学教研室,苏州215007 [3]南京医科大学附属无锡第一医院血液科,无锡214002

出　　处：《中国实验血液学杂志》2006年第5期900-902,共3页Journal of Experimental Hematology

摘　　要：本研究旨在观察葡萄糖神经酰胺合酶(glucosylceramide synthase,GCS)基因及其蛋白在人白血病细胞的表达及其与肿瘤多药耐药的关系。首先以β-actin基因为内参照物,采用RT-PCR方法分析了53例耐药/非耐药白血病患者外周血标本,并对药物敏感的HL-60细胞株和对阿霉素耐药的HL-60/ADR细胞株中GCS基因的表达水平进行检测,同时与多药耐药基因(mdr1)的表达进行比较。继而采用Westernblot法分析HL-60细胞株及HL-60/ADR细胞株中GCS蛋白及P-gp蛋白的表达情况。结果表明:白血病患者耐药组临床标本中的GCS基因扩增条带光密度相对比值明显高于非耐药组(P<0.05),且GCS基因扩增条带光密度值显著增强时往往伴有mdr1基因的高表达,两者呈正相关(P<0.01、r=0.6)。HL-60/ADR细胞株GCSmRNA及蛋白均过度表达,且明显高于HL-60细胞株,与此同时,HL-60/ADR细胞株的mdr1mRNA和P-gp的表达亦显著增高。结论:GCS可能在白血病多药耐药中起着重要作用。This study was purposed to explore the expression of glucosylceramide synthase （GCS） in human leukemia cells and its relationship with multidrug resistance. RT-PCR was used to analyze peripheral blood samples from 53 leukemia patients with multidrug resistance/non-resistance, and to detect the expression level of CJCS gene in HL-60 cells and HL-60/ADR cells, the expression level was compared with the level of mdr-1. The expressions of GCS protein and P-gp prolein in HL-60 cells and HL-60/ADR cells were assayed by Western blot analysis. The results showed that the relative optical density ratio of GCS gene amplified bands in samples of leukemia patients with drug- resistance was significantly higher than that in samples of leukemia patients with drug non-resistance group （ P 〈 0.05 ）, meanwhile the significant enhancement of optical density value of GCS gene amplified bands accompamied by high expression of mdr-1 gene. Their correlation showed positive （P 〈 0.01, r = 0.6）. The GCS mRNA and protein were overexpressed in HL- 60/ADR cells, and their expression levels were obviously higher than that in HL-60 cells, meanwhile the expression of mdr-1 mRNA and P-gp also significantly increased in HL-60/ADR cells. It is concluded that the high level of GCS in leukemia patients possibly is associated with multidrug resistance of leukemia cells.

关 键 词：葡萄糖神经酰胺合成酶 多药耐药 白血病 HL-60细胞 HL-60/ADR细胞 

分 类 号：R733.7[医药卫生—肿瘤]