Src激酶抑制剂ZD6474对白血病耐药细胞系K562／A02细胞增殖的影响及其作用机制探讨  

The role of Src kinase inhibitor ZD6474 on multi-drug resistant K562/A02 cells

作　　者：嘉红云[1] 李娟[1] 王忠英[1] 周强[1] 吴晓蔓[1] 

机构地区：[1]广州医学院第二附属医院检验科,510260

出　　处：《中华血液学杂志》2012年第3期220-224,共5页Chinese Journal of Hematology

基　　金：广州市医药卫生科技项目（201102A213018）

摘　　要：目的研究Sre激酶抑制剂ZD6474对K562及其耐药株K562／A02细胞增殖的影响及其作用机制。方法用不同浓度ZD6474处理K562、K562／A02细胞，观察细胞的增殖情况；用流式细胞术分析细胞周期的变化；用Westernblot法分析ZD6474处理前后K562／A02细胞Src激酶及其磷酸化水平。用K562和K562／A02细胞建立裸鼠皮下移植瘤模型，待肿瘤直径达到0．5～1．0cm，灌胃方法每天给予25、50及100mg／kgZD6474，生理盐水灌胃作为对照，观察ZD6474对裸鼠移植瘤的影响。并对移植瘤组织进行HE染色和免疫组化染色检测Src激酶表达（末次给药后24h）的改变。结果ZD6474呈剂量依赖的方式抑制K562和K562／A02细胞增殖，作用48h的Ic。值分别为（1．61±0．07）μmol／L和（3．22±0．21）μmol／L。μmoll／LZD6474分别作用K562和K562／A02细胞24h，可以诱导‰／G，期细胞分别由（40．2±9．4）％、（25．0±7．0）％提高至（76．3±6．1）％、（54．2±6．6）％。6μmol／LZD6474可呈时间依赖方式抑制磷酸化（P）-Src和Src激酶的表达。用灌胃方法给予ZD6474，连续给药18d，50mg／kgZD6474对K562和K562／A02移植瘤的抑制率分别为43．7％和56．3％。结论ZD6474可通过抑制Sre激酶磷酸化水平诱导K562／A02细胞凋亡；体内给药可抑制裸鼠移植瘤的生长。Objective To investigate the role of Src kinase inhibitor ZD6474 on the growth of muhidrug-resistant K562/A02 ceils and its regulatory mechanisms. Methods The possible mechanisms of drugresistance were tested by Western blot. Proliferation assays and cell cycle distribution were analyzed by WST metric analysis. Western blot were used to investigate the mechanisms of antiproliferative activity induced by tyrosine kianse inhibitor ZD6474. The in vivo anti-tumor activity was evaluated in K562, K562/A02 xenografted nude mice by administration of ZD6474 （25 - 100 mg·kg-1 . d-1, PO）. Results Compared with parental K562 cells, marked high levels of p-Src and Sre expression were detected in K562/A02 ceils. WST results showed that the ICsovalues of ZD6474 on K562 and K562/A02 after 48 hours incubation were （1.61 ±0.07 ） μmol/L and （3.22 ±0.21 ） μmol/L, respectively. ZD6474 caused an accumulation of ceils in the G0/ G1 fraction and apoptosis by inhibiting the expressions of p-Src and Src kinase. Administration of ZD6474 produced a dose-dependent inhibition of tumor growth. 50 mg/kg ZD6474 produced the growth inhibition rates of 43.7% and 56.3%, respectively in K562 and K562/A02. Conclusion Our results indicated that inhibiting Src kinase could induce K62/A02 cells apoptosis in vitro and in vivo.

关 键 词：Src激酶抑制剂 抗药性 多药 K562细胞 

分 类 号：R733.7[医药卫生—肿瘤]