尼洛替尼联合5-溴汉防己甲素逆转K562/A02细胞耐药的研究  

Study on reversal effect of nilotinib in combination with 5-BrTet on multidrug resistance of K562/A02 cell line

作　　者：陈宝安[1] 单学赟[1] 程坚[1] 王飞[1] 丁家华[1] 高冲[1] 赵刚[1] 王雪梅[2] 许文林[3] 高峰[1] 夏国华[1] Michael Schmitt 

机构地区：[1]东南大学临床医学院,中大医院血液科,南京210009 [2]东南大学生物科学与医学工程学院生物电子学国家重点实验室 [3]江苏大学附属人民医院血液科 [4]Internal Medicine, University of Rostock, Germany

出　　处：《中华血液学杂志》2010年第6期385-388,共4页Chinese Journal of Hematology

基　　金：国家自然科学基金（30740062）;高等学校博士学科点专项科研基金（20070286042）

摘　　要：目的 探讨尼洛替尼、5-溴汉防己甲素（BrTet）及其联合应用对K562/A02细胞多药耐药的逆转作用及其机制.方法 尼洛替尼、BrTet单独或联合作用于K562/A02细胞,应用MTY法检测细胞增殖抑制率、流式细胞术检测细胞凋亡率、RT-PCR检测mdr1 mRNA的表达及Western blot分析P糖蛋白（P-gp）的表达的情况.结果 5 nmol/L尼洛替尼和0.5μmol/L BrTet单独使用48 h后,柔红霉素（DNR）对K562/A02细胞的IC50分别为4.52 mg/L和5.41 mg/L,联合使用后,DNR对K562/A02细胞的IC50降为2.98 mg/L.单用DNR、尼洛替尼和BrTet均不能增加K562/A02细胞凋亡率（P〉0.05）,DNR联合尼洛替尼和BrTet后细胞凋亡率明显增高.单用5 nmol/L尼洛替尼和0.5μmol/L BrTet 48 h后,K562/A02细胞mdr1 mRNA灰度值为0.48±0.04、0.64±0.01,两者合用K562/A02细胞mdr1 mRNA灰度值下降为0.35±0.04.单用5 nmol/L尼洛替尼作用48 h,P-gp的表达水平为0.61±0.05;单用0.5μmoL/L BrTet作用48 h,P-gp的表达水平为0.52 ±0.02;两者合用K562/A02细胞P-gp的表达水平降为0.44±0.03.结论 单独应用尼洛替尼和BrTet均可部分逆转K562/A02细胞的耐药,机制可能与降低mdr1 mRNA和P-gp的表达及增加K562/A02细胞凋亡有关,并且两药联用具有明显的协同作用.Objective To investigate the reversible effect of nilotinib,BrTet（5-bromotetrandrine）and their combination on multidrug resistance cell line K562/A02 and its mechanism.Methods Cell proliferation inhibition wag assessed by MTT method and cell apoptosis by flow cytometry（FCM）.The expression of mdr1 mRNA was determined by RT-PCR,and the expression of P-gp was assessed by Western blot.Results After 48 h 5 nmol/L nilotinib or 0.5μmol/L BrTet treatment,IC50 of daunorubicin（DNR）to K562/A02 was 4.52 mg/L or 5.41 mg/L respectively;While on combinative treatment,its IC50 decreased to 2.98mg/L.Nilotinib or BrTer alone was not able to increase the DNR induced apoptosis rate of K562/A02 cell （P〉0.05）,while on combination treatment the apoptosis rate increased remarkably.After 48 h 5 nmol/Lnilotinib or 0.5 μmol/L BrTet treatment alone,gray-scale value of mdr1 mRNA was 0.48±0.04 or 0.64±0.01,respectively;while on combinative treatment the value decreased to 0.35±0.04.The P-gp expression level in K562/A02 cells was 0.61±0.05.or 0.52±0.02 when treated with 5 nmol/L nilotinib or 0.5 μmol/L BrTet alone for 48 h.but on combination treatment,the level decreased to 0.44±0.03.Conclusion Nilotinib or BrTet alone can partially reverse drug resistance of K562/A02 cells.The mechanism may be associated with the decrease of mdr1 mRNA and P-gp expression and increase of the apoptosis rate.And there is a synergistic action with these two agants in combination.

关 键 词：尼洛替尼 5-溴汉防己甲素 基因 mdrl 糖蛋白类 

分 类 号：R965[医药卫生—药理学]