【摘要】 目的探讨尿嘧啶磷酸核糖转移酶基因(UPRT)对胞嘧啶脱氨酶/5-氟胞嘧啶(CD/5-FC)自杀基因系统抗瘤作用的增强效应。方法构建重组质粒pGEX-UPRT,以电穿孔法转化婴儿双歧杆菌,筛选出阳性克隆菌,加入5-FC厌氧条件下培养24h后取其上清,加到B16-F10细胞培养液中,采用MTT法检测细胞生存率,并观察细胞形态学改变。结果阳性克隆菌可正确表达UPRT基因。与对照组比较,CD组、UPRT组及CD/UPRT组细胞存活率均明显下降;与CD组及UPRT组比较,CD/UPRT组细胞存活率明显降低。且CD/UPRT组肿瘤细胞显示出明显的损伤性改变,而CD组、UPRT组变化不如前者。结论婴儿双歧杆菌介导的UPRT基因可明显增强CD/5-FC系统对鼠黑色素瘤细胞B16-F10的杀伤作用。更多还原

【Abstract】 Objective To evaluate the UPRT-enhanced antitumor effect of CD/5-FC suicide gene system.Methods Recombined plasmid pGEX-UPRT was constructed and transferred into bifidobacterium infants by electroporation.Positive recombined bacteria was selected and identified,and then was incubated with 5-FC in anaerobic condition.The supernatant of bacteria was collected and added to the melanoma B16-F10 cells.The cell growth status was evaluated by MTT assay,and the morphologic changes of cells were observed.Results Recombinant bifidobacterium infants could express UPRT correctly.The survival rate of cells was significantly lower in CD group、UPRT group and CD/UPRT group than in control group;and the survival rate of cells was significantly lower in CD/UPRT group than in CD group and UPRT group.The cells of CD/UPRT group were remarkably damaged morphologically,and the cell growth was significantly inhibited as compared with that of other groups.Conclusion Recombinant bifidobacterium infants with UPRT gene can significantly enhance antitumor effect of CD/5-FC suicide gene system on mice melanoma B16-F10 cells.更多还原