西妥昔单抗联合低温热疗诱导CNE细胞凋亡的研究  

Apoptosis Induced by Cetuximab Combined with Hypothermia in CNE Cells

作　　者：徐寒子[1] 王仁生[1] 黄光武[2] 韦波[1] 

机构地区：[1]广西医科大学第一附属医院放疗科,广西南宁530021 [2]广西医科大学第一附属医院耳鼻喉科,广西南宁530021

出　　处：《现代生物医学进展》2009年第1期39-42,76,共5页Progress in Modern Biomedicine

基　　金：广西医疗卫生重点科研课题资助项目(重200626)

摘　　要：目的:观察西妥昔单抗联合低温热疗诱导人鼻咽癌细胞株CNE凋亡的效果,并初步探讨其可能的机制。方法:试验分对照组、单靶组、单热组及热靶组,MTT法测定西妥昔单抗对CNE细胞株48h20～30%抑制的药物浓度;以该浓度药物与低温热疗(43℃,30分钟)联合,Hoechst33258荧光染色法观察细胞凋亡形态学变化;流式细胞术检测24h、48h凋亡率;western blot检测Bax、Bcl-2、EGFR蛋白的表达。结果:以48h20～30%抑制率的药物浓度(IC20～30)为试验的工作浓度,确定西妥昔单抗对CNE细胞株的工作浓度为10ug/ml;荧光染色法观察到热靶组CNE细胞发生典型的凋亡形态学改变;流式凋亡检测显示24、48h热靶组凋亡率显著高于相应单靶组、单热组及对照组(P<0.05)。western blot检测显示各处理组较之对照组都有上调Bax、下调Bcl-2、EGFR蛋白表达的作用,以热靶组最为显著(P<0.05);热疗后EGFR蛋白表达逐渐下调。结论:西妥昔单抗联合低温热疗能显著增加CNE细胞的凋亡率,这可能与EGFR的下调与受抑,进而上调Bax、下调Bcl-2蛋白的表达促进凋亡有关。Objective： To investigate the apoptosis and its mechanism induced by cetuximab combined with hypothermia in CNE cells. Methods： The working concentration of cetuximab was defined as its inhibiting concentration （IC） （20-30%） at 48 hours action against CNE which was determined by MTT assay. Apoptosis was assayed by fluorescent Hocehst 33258 staining and flow cytometry （FCM） in four experimental groups： control group without treatment, cetuximab （T） or hyperthermia （H）（43 ℃, 30min）alone, cetuximab combined with hypothermia （TH）. The expression of Bcl- 2, Bax, EGFR proteins were determined by Western Blot. Results： A working concentration of Cetuximab at 10 μg/ml was obtained and subsequently used in all experiments. Characteristical changes of apoptosis in cells were observed in all the experient groups, especially in TH group; FCM also showed more prominent apoptosis induced by TH group than the control, T and H group. Western Blot analysis indicated that the expression of the proteins Bax has significantly increased while Bcl-2 decreased by the treatment. EGFR decreased gradually after treatment by hypothermia. Conclusion： Cetuximab combined with hypothermia synergistically increases the apoptosis in CNE cells, possibly through the Bax up-regulation and Bcl-2 down-regulation by EGFR down-regulation.

关 键 词：表皮生长因子受体 西妥昔单抗 热疗 鼻咽癌 凋亡 

分 类 号：R739.63[医药卫生—肿瘤] R730.58[医药卫生—临床医学]