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目的通过研究苯基棕榈酰胺吗啡丙醇(PPMP)和小干扰RNA(siRNA)对白血病多药耐药细胞K562/AO2葡萄糖神经酰胺合成酶(GCS)基因的调节及对凋亡相关基因的影响,以探讨GCS基因致白血病多药耐药的分子病理机制。方法应用体外细胞培养技术和RNA干扰技术观察GCS特异性的化学抑制剂PPMP及针对GCS的siRNA对K562/AO2细胞GCS基因的抑制作用,采用逆转录-聚合酶链反应(RT-PCR)技术分析GCS基因被抑制前后K562/AO2细胞中GCS基因﹑Bcl-2基因﹑Bax基因的表达水平及差异。结果PPMP和siRNA可抑制K562/AO2细胞GCSmRNA的表达,同时致相应细胞的Bcl-2基因表达下降,Bax基因则无明显变化。结论GCS可能是通过Bcl-2信号转导途径使细胞的抗凋亡能力增强,从而导致白血病细胞的多药耐药。
OBJECTIVE: To investigate the effects of phenyl palmitamide morphine propanol (PPMP) and small interfering RNA (siRNA) on the regulation of glucosylceramide synthetase (GCS) gene expression in multidrug-resistant K562 / AO2 leukemia cells and its effect on apoptosis related genes, To investigate the molecular pathogenesis of multidrug resistance caused by GCS gene in leukemia. Methods GCS-specific inhibitors of GCS and GCS-specific siRNA were used to observe the inhibitory effect of GCS in K562 / AO2 cells by using cell culture technique and RNAi technique. RT-PCR was used to analyze the effect of GCS- GCS gene was inhibited before and after K562 / AO2 cells GCS gene ﹑ Bcl-2 gene ﹑ Bax gene expression levels and differences. Results PPMP and siRNA could inhibit the expression of GCS mRNA in K562 / AO2 cells, while the expression of Bcl-2 gene in the corresponding cells decreased, while the expression of Bax gene did not change significantly. Conclusion GCS may enhance the anti-apoptotic ability of cells through Bcl-2 signal transduction pathway, leading to multidrug resistance of leukemic cells.