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通过电转化法将真核表达载体EGFPN1和pLCDSN导入减毒鼠伤寒沙门氏菌SL32 6 1中 ,经由胃管饲于C5 7BL/ 6和BALB/c小鼠。6周后接种Lewis和 4T1肿瘤细胞 ,待肿瘤增至直径为 10mm左右 ,辅以腹腔注射 5 氟胞嘧啶治疗。通过流式细胞仪、共聚焦显微镜和PCR等方法 ,在小鼠的肝脏、脾脏、小肠、肾脏、肿瘤等组织器官中均可检测到胞嘧啶脱氨酶基因的整合 ,绿色荧光蛋白在小鼠的脾脏和肿瘤中表达最强 ,其他组织表达甚弱。利用胞嘧啶脱氨酶 / 5 氟胞嘧啶系统进行治疗的小鼠肿瘤生长较其他组显著受抑 (P <0 .0 1) ,小鼠的生存期明显延长 (P <0 .0 1) ,未观察到明显的毒副作用
Eukaryotic expression vectors EGFPN1 and pLCDSN were introduced into attenuated Salmonella typhimurium SL32 61 by electroporation and administered to C57BL/6 and BALB/c mice via gastric tube. After 6 weeks, Lewis and 4T1 tumor cells were inoculated and the tumors were increased to a diameter of about 10 mm, supplemented with an intraperitoneal injection of 5-fluorocytosine. The cytosine deaminase gene integration was detected in mouse liver, spleen, small intestine, kidney, tumor and other tissues by flow cytometry, confocal microscopy, and PCR. Green fluorescent protein was detected in mice. The expression of spleen and tumor was the strongest, and the expression of other tissues was weak. The tumor growth of mice treated with cytosine deaminase / 5 fluorocytosine system was significantly suppressed compared with other groups (P < 0.01), and the survival time of mice was significantly longer (P <0.01). No obvious toxic side effects observed