【摘要】 目的观察葡萄糖神经酰胺合酶(GCS)基因在人红白血病多药耐药细胞株K562/AO2的表达及其与肿瘤多药耐药性(MDR)的关系。方法运用Alamar BlueTM多功能细胞染色法证实K562/AO2的多药耐药性;采用RT-PCR技术检测K562/AO2及其亲本K562细胞株的GCS基因、MDR1基因、bcl-2基因、bax基因的表达水平及其差异。结果(1)阿霉素(ADR)对K562/AO2细胞和K562细胞的IC50分别为75μg/ml和0.65μg/ml,耐药指数为115;长春新碱(VCR)对K562/AO2细胞和K562细胞的IC50分别为8μg/ml和0.22μg/ml,耐药指数为36。(2)K562/AO2细胞GCS基因和MDR1基因的表达明显强于K562细胞;K562/AO2细胞bcl-2基因表达强于K562细胞,而bax基因表达弱于K562细胞。结论GCS基因可能在白血病多药耐药中起着重要的作用,而细胞凋亡基因的表达异常可能是它导致肿瘤多药耐药的主要分子病理机制之一。更多还原

【Abstract】 Objective To investigate the relationship between the expression of glucosylceramide synthase(GCS) mRNA in human erythroleukemia cell line K562/AO2 and multidrug resistance of the tumor cells.Methods Alamar BlueTM multifunction cell staining method was applied to confirm the multidrug resistant of K562/AO2 cells.Using reverse transcript polymerase chain reaction(RT-PCR),the differential expressions of GCS mRNA,MDR1 mRNA,bcl-2 mRNA and bax mRNA between K562/AO2 and K562 cell lines were analyzed.Results(1) The IC50 of ADR to K562/AO2 and K562 cell line separately was 75 μg/ml and 0.65 μg/ml,the IC50 of VCR to K562/AO2 and K562 cell line separately was 8 μg/ml and 0.22 μg/ml.The drug resistance factor separately was 115 and 36.(2) K562/AO2 cells exhibited significantly higher expressions of GCS and MDR1 gene than K562 cells,and K562/AO2 cells exhibited higher expressions of bcl-2 gene than K562 cells whereas the expressions of bax gene were higher in K562 cells.Conclusion GCS gene might play an important role in MDR during human leukemia progression.Abnormal expressions of the genes in associated with cell apoptosis might be one of the main molecular pathology mechanisms of multidrug resistance caused by GCS gene.更多还原