表皮生长因子受体酪氨酸激酶抑制剂ZD1839与伊利替康联合效应的实验研究  

Experimental study of effect of epidermal growth factor receptor tyrosine kinase inhibitor ZD1839 in combination with irinotecan

作　　者：徐建明[1] 李月敏[1] 王岩[1] 赵传华[1] 袁守军[2] 杨武威[1] 李志强[1] 韩宇[1] AmaliaAzzariti AngeloParadiso 

机构地区：[1]解放军第307医院全军肿瘤中心消化肿瘤内科,北京100071 [2]军事医学科学院放射医学研究所 [3]意大利国立癌症研究院巴里医院

出　　处：《中华肿瘤杂志》2006年第8期578-582,共5页Chinese Journal of Oncology

基　　金：全军"十一五"科技攻关资助项目(06g140)

摘　　要：目的探讨ZD1839与伊利替康的活性代谢产物7-乙基-10羟基-喜树碱(SN38)联合的最佳方案及其机制。方法以药物联合效应测定方法,评价ZD1839和SN38不同给药顺序对人结肠癌细胞HT-29和LoVo的抑制作用;以Western blot和免疫共沉淀方法,分析ZD1839与化疗不同联合方案对各自靶蛋白及其下游分子表达的影响;以流式细胞仪测定不同联合方案对细胞周期的影响;以组蛋白相关的DNA碎片分析,比较不同方案对细胞凋亡指数的影响。结果先SN38后ZD1839的序贯给药方案表现出明显的协同作用;反之,则表现为拮抗作用。SN38明显抑制细胞拓扑异构酶-I (Topo-I)活性;ZD1839不影响表皮生长因子受体(EGFR)的表达,但能抑制EGFR的磷酸化。与SN38单药相比,SN38联合ZD1839对Topo-I的抑制无增强;与单药ZD1839相比,联合方案对EGFR、MAPK磷酸化的抑制作用无增强,但ZD1839、SN38同时给药,和先SN38后ZD1839序贯给药对AKT的抑制有所增强。同时,联合方案对细胞周期分布的改变影响明显。ZD1839可明显维持化疗诱导的DNA损伤和细胞凋亡。结论先SN38后ZD1839序贯给药可能是治疗结肠癌的最佳联合模式。Objective To assess the optimal regimen and its mechanism of ZD1839 in combination with SN38, the active metabolite of irinotecan （CPT-11）, in the colon cancer cell lines HT-29 and LoVo. Methods Chou and Talalay method was used to analyze the combination effects of sequencing of ZD1839 and SN38. Western blotting and immunoprecipitation were used to determine the effects of ZD1839 and/or SN38 on their targeted enzymes and downstream markers. Apoptosis was assayed by analyzing histoneassociated DNA fragment. Results Sequential SN38 followed by ZD1839 produced a synergistic effect. In contrast, SN38 following ZD1839 exhibited an antagonist effect. SN38 markedly inhibited topoisomerase Ⅰ （Topo- Ⅰ ） activity. ZD1839 did not alter epidermal growth factor receptor （EGFR） expression, but resulted in a complete inhibition of EGFR phosphorylation. Sequential ZD1839 followed by SN38 did not show any enhanced inhibition effect on Topo-Ⅰ activity, phosphorylation of EGFR and one of its downstream markers MAPK. However, simultaneous SN38 plus ZD1839, and sequential SN38 followed by ZD1839 administrations showed modest inhibition effect on EGFR＇s another downstream marker AKT. The combination schedules also showed prominent influence on cell cycle distribution. ZD1839 maintained SN38induced DNA damage and apoptosis. Conclusion Sequential SN38 followed by ZD1839 may be a favorable combination schedule.

关 键 词：ZD1839 伊利替康 SN38 结肠癌细胞系HT-29 结肠癌细胞系LoVo 

分 类 号：R96[医药卫生—药理学]