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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。
Objective To investigate the expression of glucosylceramide synthase (GCS) gene in human erythroleukemia multidrug resistant cell line K562 / AO2 and its relationship with multidrug resistance (MDR). Methods The multidrug resistance of K562 / AO2 cells was confirmed by Alamar BlueTM multifunctional cell staining. The expression of GCS, MDR1, bcl-2 and bax genes in K562 / AO2 and its parental K562 cell lines was detected by RT-PCR The expression level and its difference. Results (1) IC50 of Adriamycin (ADR) on K562 / AO2 cells and K562 cells were 75μg / ml and 0.65μg / ml, respectively. The resistance index was 115. VCR was more effective on K562 / AO2 cells and K562 The IC50 of the cells were 8μg / ml and 0.22μg / ml respectively, and the resistance index was 36. (2) The expression of GCS gene and MDR1 gene in K562 / AO2 cells was stronger than that in K562 cells. The expression of bcl-2 gene in K562 / AO2 cells was stronger than that in K562 cells, while the expression of bax gene was weaker than that in K562 cells. Conclusions GCS gene may play an important role in multidrug resistance of leukemia. Abnormal expression of apoptosis gene may be one of the major molecular pathological mechanisms that lead to multidrug resistance of tumor.