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目的探讨葡萄糖神经酰胺合酶(GCS)基因在细胞株K562/AO2和K562中的表达与白血病多药耐药(MDR)的关系。方法RT-PCR技术检测K562/AO2及K562细胞株的GCS和Bcl-2基因的表达;ELISA法检测两细胞株中Bcl-2及Caspase-3的表达;AlamarBlueTM细胞染色法分析K562/AO2细胞经苯基棕榈酰胺吗啡丙醇(PPMP)处理后多药耐药性的变化。结果K562/AO2细胞的GCS基因、Bcl-2基因和Bcl-2蛋白表达明显强于K562细胞(P<0.01),Caspase-3则相反(P<0.05)。K562/AO2细胞经25μmol/LPPMP处理后,GCS基因表达抑制,阿霉素(ADR)对其半数抑制浓度(IC50)明显降低。结论GCS基因可能在白血病MDR形成过程中起着重要作用,其机制可能为细胞凋亡相关基因的表达异常。PPMP通过抑制GCS活性有效逆转K562/AO2细胞的多药耐药性。
Objective To investigate the relationship between the expression of glucosylceramide synthase (GCS) gene in cell lines K562 / AO2 and K562 and multidrug resistance (MDR) in leukemia. Methods The expression of GCS and Bcl-2 genes in K562 / AO2 and K562 cell lines were detected by RT-PCR. The expressions of Bcl-2 and Caspase-3 in the two cell lines were detected by ELISA. The cell cycle of K562 / AO2 cells was analyzed by AlamarBlueTM staining Changes of multidrug resistance after treatment with phenyl palmitamide morphine propanol (PPMP). Results The expressions of GCS, Bcl-2 and Bcl-2 in K562 / AO2 cells were significantly higher than those in K562 cells (P <0.01) and Caspase-3 (P <0.05). GCS gene expression was inhibited in K562 / AO2 cells treated with 25μmol / L LPMP, and the IC50 of Adriamycin (ADR) was significantly decreased. Conclusions GCS gene may play an important role in the development of MDR in leukemia, which may be caused by the abnormal expression of apoptosis-related genes. PPMP effectively reversed the multidrug resistance of K562 / AO2 cells by inhibiting GCS activity.