α干扰素增强骨肉瘤U2OS细胞对足叶乙甙的敏感性  

IFNα enhances chemosensitivity of human osteosarcoma U2OS cells to VP16

作　　者：原向伟[1] 廖威明[2] 朱孝峰[3] 黄秀芳[4] 康焱[2] 雷磊[2] 黄保丁[2] 

机构地区：[1]中山大学附属江门医院骨科,529030 [2]中山大学附属第一医院骨科,广州510080 [3]华南肿瘤学国家重点实验室,广州510060 [4]中山大学附属江门医院病理科,529030

出　　处：《中华关节外科杂志（电子版）》2007年第5期294-296,共3页Chinese Journal of Joint Surgery(Electronic Edition)

基　　金：国家自然科学基金资助项目(30070766)

摘　　要：目的探讨α干扰素对人骨肉瘤U2OS细胞足叶乙甙敏感性的作用及其机制,为提高骨肉瘤化疗的敏感性探索新的治疗方法。方法应用MTT法测定IFNα和VP16单用以及联用对U2OS细胞的细胞毒作用;Hoechst33258荧光染色检测药物对U2OS细胞凋亡的影响;Westernblot法检测Caspase-3、8、9的表达和活化。结果IFNα在50U/ml、500U/ml、5000U/ml等剂量处理48h、72h、96h对U2OS细胞无毒性作用,却明显增强了VP16(2.5μg/ml)诱导的细胞毒作用;与单药组相比,IFNα(5000U/ml)与VP16(2.5μg/ml)联用72h后U2OS细胞出现更为明显的凋亡特征性形态变化;并且联合用药组的凋亡关键酶Caspase-3、8、9均发生明显断裂活化。结论IFNα能够增强VP16诱导的骨肉瘤U2OS细胞毒作用和凋亡,且与Caspase级联活化有关,二者联合应用可能成为提高骨肉瘤化疗敏感性的有效途径。Objective To study effect of IFNα on VP16 chemosensitivity in human osteosarcoma U2OS cells with its molecular mechanisms to provide evidences for increasing chemosensitivity of osteosar- coma. Methods Cell viability was evaluated using MTT assay. Apoptosis was studied using Hoechst 33258 staining. Activation of Caspase-3,8,9 was detected by Western blot. Results IFNct （50 U/ml, 500 U/ml, 5000 U/ml） treatment for48 h, 72 h and 96 h did not induce cytotoxicity but greatly enhanced VP16-induced cytotoxicity in U2OS cells. Compared with other groups, the combination of IFNct （5000 U/ ml） and VP16 （2. 5 μg/ml） induced more obvious apoptotic morphological changes. Caspase-3,8,9 were cleaved to yield active fragment following IFNα/VP16 combination in U2OS cells, compared with other groups. Conclusion IFNα enhances VP16-induced cytotoxicity and apoptosis in human osteosarcoma U2OS cells by activation of caspases cascade. This work implies that rational combination with IFNα and chemotherapy might be a useful strategy for treatment of osteosarconma.

关 键 词：Α干扰素 足叶乙甙 骨肉瘤 凋亡 半胱氨酸天门冬氨酸蛋白酶 

分 类 号：R738[医药卫生—肿瘤]