葡萄糖神经酰胺合酶及白血病细胞耐药关系.docVIP

葡萄糖神经酰胺合酶及白血病细胞耐药关系 　 ; 作者：谢平，葛素梅，沈云峰，顾中华，穆会君，张滨 毕业论文 【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（Plt;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（Plt;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。 毕业论文 【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 毕业论文 　　Relation Between Glucosylceramide Synthase and Multidrug Resistance in Leukemia Cells 毕业论文 　　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 (Plt;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 (Plt;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 GC