高压氧联合阿霉素对白血病K562/A02细胞凋亡的影响

本研究探讨0.2MPa高压氧(HBO)联合阿霉素对白血病多药耐药细胞K562/A02细胞凋亡的影响。利用流式细胞术法和透射电子显微镜检测细胞凋亡和caspase-3表达,用定量qReal-time PCR法检测HIF-1α、BCL-2和BAXmRNA表达水平,运用caspase-8试剂盒检测caspase-8的活性,Western blot法检测P-gp活性。结果表明:0.2MPa HBO联合阿霉素组细胞凋亡率高于ADM组(47.36±3.87)%vs(28.51±1.09)%],差异有统计学意义(p<0.05)。0.2MPa HBO联合阿霉素组细胞HIF-1αmRNA、P-gp及BCL-2的蛋白水平低于ADM组,差异有统计学意义(p<0.05)。0.2MPa HBO联合阿霉素组细胞的BAX、caspase-3及caspase-8蛋白活性均高于ADM组,差异有统计学意义(p<0.05)。结论 :0.2MPa HBO联合阿霉素可逆转耐药细胞株K562/A02对阿霉素的耐药,提高细胞凋亡率,其机制可能与HIF-1α下调P-gp和BCL-2的表达以及提高caspase活性有关。

Effects of hyperbaric oxygen combined with adriamycin in inducing apoptosis of K562/A02 cells

This study was purposed to explore the effects of hyperbaric oxygen （HBO） combined with adriamycin （ADM） on inducing apoptosis of multidrug resistant cells line K562/A02. The cell apoptosis and expression of caspase- 3 activity were analyzed by flow cytometry and transmission electron microscopy; the expression levels of HIF-1 α, BCL- 2 and BAX mRNA were detected by quantitative real time PCR; the caspase 8 activity was determined by using caspase 8 kit; the expression level of P-gp was detected by Western blot. The results showed that the apoptosis rate of K562/A02 cells in combination group （0.2 MPa HBO ＋ ADM） was higher than that in ADM group （47.36 ±3.87） % vs （28.51 ± 1.09） % ], the difference was statistical significant （p 〈 0.05） ; the expression levels of HIF-let mRNA, P- gp and BCL-2 in combination group were lower than those in ADM group, there were significant differences （p 〈 0.05） ; the activities of BAX, caspase 3 and caspase 8 proteins in combination group were higher than those in ADM group, the difference was statistical significant （p 〈 0.05）. It is concluded that 0.2 MPa HBO combined with ADM can reverse the drug-resistance of K562/A02 cells to ADM, enhance the apoptosis rate of cells. The molecular mechanism may be related with downregulation of P-gp and BCL-2 expression, and up-regulation of caspase-3 and caspase-8 activities by HIF-1α.