CD-TK双自杀基因系统对腺样囊性癌ACC-2细胞放疗增敏的作用  

Radiosensitization by double suicide gene system in ACC-2 cell

作　　者：张娜[1] 张东升[1] 韩俊庆[2] 张世周[1] 刘桂军[1] 魏奉才[3] 张捷[4] 牟文丽[4] 

机构地区：[1]山东大学附属省立医院口腔颌面外科,山东济南250021 [2]山东大学附属省立医院肿瘤中心,山东济南250021 [3]山东大学齐鲁医院口腔颌面外科,山东济南250012 [4]山东大学附属省立医院科研中心.山东济南250021

出　　处：《上海口腔医学》2009年第2期183-188,共6页Shanghai Journal of Stomatology

基　　金：山东省自然科学基金(Z2003C01)~~

摘　　要：目的:探讨前体药物及CD-TK双自杀基因系统对人腺样囊性癌(ACC-2)细胞的放疗增敏作用。方法:重组真核表达质粒pIRES-CD及pIRES-TK经电穿孔法共转染ACC-2细胞,以400μg/mL的G418筛选10d,获得稳定表达CD及TK基因的ACC-2细胞,提取该细胞的总RNA,RT-PCR检测CD、TK基因的表达;将阳性克隆的ACC-2细胞分别在有氧及乏氧条件下给予不同剂量(0、2、4、6、8、10Gy)X线照射及前体药物干预,通过细胞克隆形成实验,观察双自杀基因及前体药物在有氧及乏氧条件下对ACC-2细胞的放疗增敏作用。采用SPSS11.5软件包对数据进行多因素方差分析。结果:RT-PCR检测到ACC-2/CD-TK中CD、TK基因的表达;随X线照射剂量的增加,各组细胞放疗后克隆形成率均明显降低;有氧条件下,X线照射ACC-2/CD-TK+前体药物组细胞存活分数均较相同剂量X线照射ACC-2及ACC-2/CD-TK细胞组低(P<0.05);乏氧条件下,X线照射ACC-2/CD-TK+前体药物组细胞存活分数均较相同剂量X线照射ACC-2及ACC-2/CD-TK细胞组低(P<0.05);相同照射剂量下,各相应细胞组在有氧条件下细胞存活分数较乏氧条件下低。结论:CD-TK双自杀基因及其前体药物的应用,可以提高ACC-2细胞放疗敏感性及X线放射治疗对ACC-2细胞的杀伤作用。PURPOSE： To investigate the radiosensitization by prodrug and CD-TK double suicide gene therapy system in adenoid cystic carcinoma cells （ACC-2）. METHODS： The eukaryotic expression plasmids pIRES-CD and pIRES-TK were introduced into ACC-2 cells by electroporation. Then ACC-2 cells stably expressing CD and TK gene were obtained by 10-day positive selection with 400 μg/mL G418 . The total RNA was extracted and the expression of the CD and TK gene in transfected ACC-2 cells was identified by RT-PCR. The positive transfected ACC-2 cells were treated with radiotherapy of different dose （0,2,4,6,8,10Gy） and prodrug system in aerobic and anoxic condition. Then cell clone formation assay was used to study the radiosensitization by CD-TK double suicide gene therapy and prodrug system in ACC-2.The data was analyzed by multiple factor ANOVA using SPSS11.5 software package. RESULTS： RT-PCR analysis demonstrated that CD and TK genes were effectively expressed in ACC-2 cells. With the increased of X-ray dose, the colony forming rate dropped significantly after radiotherapy. In aerobic condition, the survival fraction of group ACC-2/CD-TK＋prodrug were significantly lower than that of group ACC-2 and group ACC-2/CD-TK with the same dose （P〈0.05）. In anoxie condition, the survival fraction of group ACC-2/CD-TK＋pro-drug was significantly lower than that of experimental group ACC-2 and group ACC-2/CD-TK with the same dose （P〈0.05）. The colony forming rate in aerobic condition was significantly lower than that in anoxic condition of the same cell group and dose. CONCLUSION： The radiosensitivity and the killing effect of X ray to ACC-2 cells can be increased by CD-TK double suicide gene therapy and the prodrug system.Supported by Natural Science Foundation of Shandong Province（Grant No.Z2003C01）.

关 键 词：自杀基因治疗 前体药物 放射疗法 克隆形成率 腺样囊性癌 

分 类 号：R739.8[医药卫生—肿瘤]