论文部分内容阅读
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。
Objective: To investigate the radiosensitization effect of prodrug and CD-TK suicide gene system on human adenoid cystic carcinoma (ACC-2) cells. METHODS: ACC-2 cells were co-transfected with recombinant eukaryotic expression plasmids pIRES-CD and pIRES-TK by electroporation. ACC-2 cells were stained with G418 at 400 μg / mL for 10 days to obtain ACC-2 cells stably expressing CD and TK genes. The total RNA was extracted and the expression of CD and TK genes was detected by RT-PCR. The positive clones of ACC-2 cells were treated with different doses (0, 2, 4, 6, 8 and 10 Gy) Line irradiation and prodrug intervention. Through cell clone formation experiment, we observed the radiosensitization effect of double suicide genes and prodrugs on ACC-2 cells under aerobic and hypoxic conditions. Data were analyzed by multivariate analysis of variance using SPSS 11.5 software package. Results: The expression of CD and TK genes in ACC-2 / CD-TK was detected by RT-PCR. The colony formation rate of all the groups was significantly decreased with the increase of X-ray dose. Under aerobic conditions, The cell viability of ACC-2 / CD-TK + prodrug group was lower than that of ACC-2 and ACC-2 / CD-TK cells treated with the same dose of X-ray irradiation -2 / CD-TK + prodrug group were significantly lower than that of ACC-2 and ACC-2 / CD-TK cells treated with the same dose of X-ray irradiation Cell survival scores were lower under aerobic conditions than under hypoxic conditions. Conclusion: The application of CD-TK double suicide genes and their prodrugs can enhance the radiosensitivity of ACC-2 cells and the killing effect of radiotherapy on ACC-2 cells.