含有人端粒酶反转录酶的溶瘤腺病毒Ad11联合异环磷酰胺对人骨肉瘤细胞的杀伤作用及其机制  

Killing effect of oncolytic adenovirus Ad11 containing human telomerase reverse transcriptase combined with ifosfamide on human osteosarcoma cells and possible mechanism

作　　者：张岩[1] 张彬[1] 卢新昌[1] 张翼[1] 刘宏建[1] 李甲振[1] Zhang Yan;Zhang Bin;Lu Xinchang;Zhang Yi;Liu Hongjian;Li Jiazhen(Department of Orthopedic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052, China)

机构地区：[1]郑州大学第一附属医院骨科,450052

出　　处：《中华实验外科杂志》2018年第12期2292-2294,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨溶瘤腺病毒Ad11联合异环磷酰胺对人骨肉瘤细胞的杀伤作用及机制。方法体外培养人骨肉瘤细胞株MG63,采用细胞毒性检测法(MTS法)检测Ad11的50%最大效应浓度(EC50)和异环磷酰胺的25%抑制浓度(IC25)和50%抑制浓度(IC50)。使用MTS法分别检测对照组、Ad11组[转染复数(MOI)=5]、异环磷酰胺组1(IC25=15μmol/ml)、异环磷酰胺组2(IC50=25μmol/ml)、联合组1(Ad11+IC25)、联合组2(Ad11+IC50)的细胞活性。Westernblot技术检测对照组和实验组的自噬相关蛋白Beclin-1和LC3A/B的表达。结果MTS实验显示Ad11处理骨肉瘤细胞,其EC50值为5.26MOI,联合异环磷酰胺后对骨肉瘤细胞MG63具有协同杀伤作用。Westernblot结果显示Ad11联合异环磷酰胺处理细胞时,异环磷酰胺引起的细胞自噬作用受到抑制。结论Ad11对骨肉瘤细胞株MG63具有杀伤作用,联合异环磷酰胺时杀伤作用增强,该协同作用可能是由于Ad11减弱了MG63细胞的保护性自噬作用。Objective To study the killing effect of oncolytic adenovirus Ad11 combined with ifosfamide on human osteosarcoma cells and the possible mechansim, and provide cytological basis for oncolytic adenovirus Ad116 in the treatment of osteosarcoma. Methods The human osteosarcoma cell line MG63 was cultured in vitro, and the 50% maximal effect concentration (EC50) of Ad11 and the 25% inhibitory concentration (IC25) and 50% inhibitory concentration (IC50) of adrephosphamide were detected by cytotoxicity assay (MTS method). The MTS method was used to detect the cell viability of the control group, Ad11 group [multiplicity of infection (MOI)=5], ifosfamide group 1 (IC25=15 μmol/ml), ifosfamide group 2 (IC50=25 μmol/ml), combined group 1 (Ad11+ IC25) and combined group 2 (Ad11+ IC50). Western blotting was used to detect the expression of autophagy-related proteins Beclin-1 and LC3A/B in the control and experimental groups. Results The MTS assay showed that Ad11 treatment of osteosarcoma cells had an EC50 value of 5.26 MOI, and combined with ifosfamide had synergistic killing effect on osteosarcoma cell MG63. Western blotting results showed that autophagy induced by ifosfamide was inhibited when Ad11 was treated with ifosfamide. Conclusion Ad11 has a killing effect on osteosarcoma cell line MG63. The synergistic effect is enhanced when Ad11 is combined with ifosfamide. The synergistic effect may be due to the weakened protective autophagy of MG63 cells.

关 键 词：溶瘤腺病毒 Ad11 骨肉瘤 自噬 异环磷酰胺 

分 类 号：R738.1[医药卫生—肿瘤]