论文部分内容阅读
目的探索伊马替尼(IM)联合使用17-丙烯胺基-17-去甲氧基格尔德霉素(17-AAG)增加慢性粒细胞白血病(CML)细胞株K562和K562A02凋亡率的机制。方法采用四甲基偶氮唑蓝法观察细胞增殖抑制率,Annexin V/PI双染色法检测细胞凋亡率,逆转录-聚合酶链反应法检测Bcr-abl、MDR1、Bcl-2 mRNA的表达及Western blot法检测Bcr-abl、p-gp、Bcl-2蛋白的表达。结果0.31μg·mL-1伊马替尼联合1.25μg·mL-117-丙烯胺基-17-去甲氧基格尔德霉素分别作用于K562和K562A02细胞48 h,增殖抑制率分别为(70.78±1.01)%和(39.27±1.52)%,凋亡率为(48.74±2.51)%和(45.0±0.4)%。2.5μg·mL-1伊马替尼联合10μg·mL-117-丙烯胺基-17-去甲氧基格尔德霉素作用于K562A02细胞48 h,增殖抑制率达(65.63±0.93)%。逆转录-多聚酶链反应和Western blot结果显示,联合用药后K562细胞的Bcr-abl和Bcl-2基因显著降低,MDR1呈弱表达,无明显变化。而K562A02细胞中,Bcr-abl、Bcl-2和MDR1基因的表达均有明显下调(P<0.05)。结论伊马替尼联合17-丙烯胺基-17-去甲氧基格尔德霉素可有效抑制K562和K562A02细胞增殖和诱导凋亡,两者联合应用具有克服伊马替尼耐药的作用。
Objective To explore the effects of imatinib (IM) combined with 17-Amylamino-17-demethoxygeldanamycin (17-AAG) on the apoptosis of K562 and K562A02 cells in chronic myeloid leukemia (CML) mechanism. Methods The inhibitory rate of cell proliferation was detected by MTT method. The apoptosis rate was detected by Annexin V / PI double staining. The expression of Bcr-abl, MDR1 and Bcl-2 mRNA was detected by reverse transcription polymerase chain reaction Western blot was used to detect the expression of Bcr-abl, p-gp and Bcl-2. Results The inhibitory rates of imatinib combined with 1.25μg · mL-117-acrylamido-17-demethoxygeldanamycin on K562 and K562A02 cells for 48 h were ( 70.78 ± 1.01% and 39.27 ± 1.52%, respectively. The apoptotic rates were (48.74 ± 2.51)% and (45.0 ± 0.4)%, respectively. K562A02 cells were treated with 2.5μg · mL-1 imatinib and 10μg · mL-117-acrylamido-17-demethoxygeldanamycin for 48 h. The proliferation inhibition rate was (65.63 ± 0.93)%. The results of RT-PCR and Western blot showed that the Bcr-abl and Bcl-2 genes in K562 cells were significantly decreased and the expression of MDR1 was weakly expressed with no significant changes. However, the expression of Bcr-abl, Bcl-2 and MDR1 genes in K562A02 cells were significantly down-regulated (P <0.05). Conclusions Imatinib combined with 17-acrylamido-17-demethoxygeldanamycin effectively inhibits the proliferation and induces apoptosis of K562 and K562A02 cells, and the combination of them can overcome the drug resistance of imatinib .