Zhang Wei, Chen Bao-An, Bao Wen, Cheng Jian, Sun Yan, Cheng Jie, Zhang Min, Zhang Xiao-Min, Chen Yan
Department of Pathophysiology, Southeast University Medical College, Nanjing 210009, Jiangsu Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Jun;19(3):621-5.
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-1α, 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-1α 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 down-regulation of P-gp and BCL-2 expression, and up-regulation of caspase-3 and caspase-8 activities by HIF-1α.
本研究旨在探讨高压氧（HBO）联合阿霉素（ADM）对多药耐药细胞株K562/A02凋亡的诱导作用。采用流式细胞术和透射电子显微镜分析细胞凋亡及半胱天冬酶-3活性表达；通过定量实时PCR检测HIF-1α、BCL-2和BAX mRNA的表达水平；使用半胱天冬酶8试剂盒测定半胱天冬酶8活性；通过蛋白质免疫印迹法检测P-糖蛋白（P-gp）的表达水平。结果显示，联合组（0.2MPa HBO + ADM）K562/A02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）；联合组HIF-1α mRNA、P-gp和BCL-2的表达水平低于ADM组，差异有统计学意义（p < 0.05）；联合组BAX、半胱天冬酶3和半胱天冬酶8蛋白的活性高于ADM组，差异有统计学意义（p < 0.05）。结论：0.2MPa HBO联合ADM可逆转K562/A02细胞对ADM的耐药性，提高细胞凋亡率。其分子机制可能与下调P-gp和BCL-2表达，以及HIF-1α上调半胱天冬酶-3和半胱天冬酶-8活性有关。
