汉防己甲素逆转白血病细胞株K562/A02耐药的机制

doi:吉林省科技厅科研基金资助课题（20050412-3

汉防己甲素逆转白血病细胞株K562/A02耐药的机制

赵殿凤¹，刘子玲¹，马宁¹，张永峰¹，屈蓓蓓¹，王志虹²，李志毅²

1.吉林大学第一医院肿瘤治疗中心，吉林长春 130021；2.成都康弘药业集团，四川成都 610036

收稿日期:2008-03-31 修回日期:1900-01-01 出版日期:2009-01-28 发布日期:2009-01-28
通讯作者: 刘子玲

Mechanism of tetrandrine in reversion of multidrug resistance of leukemic cell line K562/A02

ZHAO Dian-feng¹,LIU Zi-ling¹,MA Ning¹,ZHANG Yong-feng¹,QU Bei-bei¹,WANG Zhi-hong²,LI Zhi-yi²

1.Center of Tumor Treatment,First Hospital,Jilin Univercity,Changchun 130021,China;2.Nakasone Pharmaceutical Group,Chengdu 610036,China

Received:2008-03-31 Revised:1900-01-01 Online:2009-01-28 Published:2009-01-28
Contact: LIU Zi-ling

摘要/Abstract

摘要： 目的:研究汉防己甲素（TTD）对白血病细胞株K562/A02多药耐药(MDR)逆转的机理。方法：以白血病细胞系K562及其耐药细胞系K562/A02为TTD作用的靶细胞。细胞水平检测实验分5组（K562组、K562/A02组、K562+ADM组、K562/A02+ADM组和K562/A02+TTD+ADM组），采用MTT法检测TTD对K562和K562/A02细胞的非细胞毒性剂量，流式细胞术检测细胞内阿霉素(ADM)的浓度，基因、酶学、蛋白水平检测实验分3组（K562组、K562/A02组和K562/A02+TTD组），采用RT-PCR法检测mdr1 mRNA的表达，免疫细胞化学方法检测谷胱甘肽S转移酶π（GST-π）和拓扑异构酶Ⅱ（Topo Ⅱ）的表达水平，Western-blotting法检测P-糖蛋白（P-gp）和bcl-2表达。结果：1.562 5 mg•L^-1的TTD处理K562/A02细胞后，细胞内ADM的浓度较单用ADM组明显提高（P<0.01）；与空白对照组比较，K562/A02细胞内mdr1 mRNA/P-gp的表达量减少（P<0.01）；GST-π和TopoⅡ表达无明显变化；凋亡抑制基因bcl-2的表达量减少（P<0.01）。结论:TTD主要通过增加细胞内ADM浓度，下调mdr1/P-gp和bcl-2表达逆转耐药。

关键词: 白血病, 汉防己甲素

Abstract: Abstract:Objective To investigate the reversal mechanism of tetrandrine (TTD) on multidrug resistance(MDR) in leukemia cell line K562/A02.Methods Human leukemia cell line K562 and multidrug-resistant cell line K562/A02 were used as the target cells.There were five groups in the examination of cellular level(K562 group,K562/A02 group,K562+ADM group,K562/A02+ADM group and K562/A02+TTD+ADM group）.The non-cytotoxicity doses to cell lines K562 and K562/A02 of TTD were got by MTT assay.Using flow cytometry (FCM) assay to examine the intracellular ADM concentration.There were three groups in the examination of genic,zymologic and protein levels(K562 group,K562/A02 group and K562/A02+TTD group）.The mRNA expression of MDR was measured by fluorescent quantitative reverse transcriptase polymerase chain reaction(RT-PCR).The expression levels of glutathione-S-transferase (GST-π) and topoisomerase Ⅱ(Topo Ⅱ) were determined by immunohistochemical technique.The expressions of P-glyco-protein(P-gp) and bcl-2 were observed by Western-blotting.Results The intracellular concentration of ADM in K562/A02 cells after treated with 1.562　5 mg•L^-1 TTD was higher than that in ADM groups(P<0.01).Compared with blank group,the expression of mdr1 mRNA/ P-gp in K562/A02 cells reduced(P<0.01),the expressins of GST-π and Topo Ⅱ had no change,and the expression of bcl-2 reduced（P<0.01）.Conclusion TTD can reverse the MDR of K562/A02 cell line by inceasing intracellular concentration of ADM and reducing the expressions of mdr1 mRNA/ P-gp and bcl-2 in K562/ A02 cells.

Key words: leukemia, tetrandrine (TTD)

中图分类号:

R733.7

赵殿凤，刘子玲，马宁，张永峰，屈蓓蓓，王志虹，李志毅. 汉防己甲素逆转白血病细胞株K562/A02耐药的机制[J]. J4, 2009, 35(1): 100-103.

ZHAO Dian-feng,LIU Zi-ling,MA Ning,ZHANG Yong-feng,QU Bei-bei,WANG Zhi-hong,LI Zhi-yi. Mechanism of tetrandrine in reversion of multidrug resistance of leukemic cell line K562/A02[J]. J4, 2009, 35(1): 100-103.

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