葡萄糖神经酰胺合酶与白血病细胞耐药的关系_药学论文.docVIP

葡萄糖神经酰胺合酶与白血病细胞耐药的关系_药学论文 葡萄糖神经酰胺合酶与白血病细胞耐药的关系_药学论文 作者：谢平，葛素梅，沈云峰，顾中华，穆会君，张滨【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Western blot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（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（0.01、r＝ 0.6）。HL-60/ADR细胞株GCS mRNA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mdr1 mRNA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。 【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 　　Relation Between Glucosylceramide Synthase and Multidrug Resistance in Leukemia Cells 　　AbstractThis 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 GCS 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 (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 (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 significanfly increased in HL-60/ADR cells. It is concluded that the high level of GCS in leukemia patients possibly is associate