Thapsigargin逆转K562/A02细胞多药耐药性的实验研究  

Reversal of Multidrug Resistance by Thapsigargin in K562/A02 Cells

作　　者：冯献启[1] 刘芳[1] 邹萍[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液科,武汉430022

出　　处：《华中科技大学学报（医学版）》2005年第2期175-178,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金资助项目 (No 30370595)

摘　　要：目的　探讨内质网Ca2+ ATP酶抑制剂thapsigargin对白血病多药耐药细胞株K562/A02 化疗敏感性的影响。方法　用MTT比色法检测K562/A02细胞耐药性、thapsigargin的增殖抑制活性及其对K562/A02细胞化疗敏感性的影响; 丫啶橙(AO) /溴化乙锭(EB) 染色荧光显微镜观察thapsigargin处理后K562/A02 细胞形态改变; 流式细胞仪(FCM) 检测P 糖蛋白(P- gp) 表达; 荧光显微镜检测P-gp功能。结果　①thapsigargin对K562 和K562/A02细胞的增殖抑制活性呈剂量和时间依赖性, K562/A02 细胞较K562 细胞对thapsigargin更敏感, thapsigargin诱导K562/A02细胞呈现典型的凋亡细胞形态学改变。②K562/A02细胞对阿霉素(ADM)、柔红霉素(DNR)、长春新碱(VCR)、依托泊苷(VP- 16)、高三尖杉酯碱(HHT) 和米托蒽醌(MXT) 均出现不同程度的耐药性; thapsigar-gin抑制K562/A02细胞P- gp功能而对其表达无影响, thapsigargin 能增加K562/A02 细胞对ADM、DNR、VCR、VP- 16、HHT和MXT的化疗敏感性。结论　Thapsigargin能诱导白血病多药耐药细胞株K562/A02 细胞凋亡并能部分逆转K562/A02的多药耐药性; thapsigargin的多药耐药逆转功能可能与其凋亡诱导作用和抑制P gp功能有关。Objective To study the effect of thapsigargin on chemotherapeutic sensitivity in multidrug-resistant leukemia cell line K562/A02 induced by ADM.Methods Multidrug resistance of K562/A02 cells,the cell-proliferating inhibitory activity of thapsigargin and the effect of thapsigargin on chemotherapeutic sensitivity were assessed by using MTT assay. Morphological change of K562/A02 cells treated with thapsigargin was examined with AO/EB fluorescent staining under fluorescent microscopy. The P-gp expression of K562/A02 cells was detected by flow cytometry and P-gp function was evaluated with adriamycin (ADM),daunorubicin (DNR) and Rh123 accumulation in K562/A02 cells by fluorescent microscopy.Results Thapdigargin inhibited the proliferation of K562/A02 cells in a dose- and time-dependent manner. K562/A02 cells were more sensitive to thapsigargin than their parent K562 cells. Thapsigargin induced typical apoptosis of K562/A02 cells showed by typical apoptotic morphological changes. K562/A02 cells were resistant to ADM,DNR,vincristine (VCR),etoposide (VP-16),homoharringtonine (HHT) and mitoxantrone (MXT) to varying degrees. Thapsigargin significantly inhibited P-gp function of K562/A02 cells,but not the P-gp expression,and enhanced the chemotherapeutic sensitivity of K562/A02 cells to above conventional chemotherapeutic agents.Conclusion Thapsigargin induces growth inhibition and apoptosis of multidrug-resistant leukemia cell line K562/A02 and partially reverse the resistance of the cells to conventional chemotherapeutic agents. The reversal action of thapsigargin can be associated with apoptosis-induced effect and inhibition of P-gp function.

关 键 词：THAPSIGARGIN 白血病 K562 A02细胞 多药抗药性 P-糖蛋白 

分 类 号：R733.7[医药卫生—肿瘤]