论文部分内容阅读
目的 探讨以减毒沙门菌作为口服基因治疗载体的可行性。方法 通过电转化法将真核表达载体pCMVhIL 12、pCMVhGM CSF、EGFPN1导入减毒鼠伤寒沙门菌SL32 6 1中 ,经由胃管喂给BALB c和C5 7BL 6小鼠。 6周后分别用 4T1乳腺癌细胞和Lewis肺癌细胞进行攻击。通过流式细胞仪、共聚焦显微镜检测绿色荧光蛋白在小鼠各组织中的表达 ,通过PCR和ELISA法检测IL 12、GM CSF基因的整合和表达情况。并考察肿瘤的受抑情况和小鼠的生存期。结果 在小鼠的肝、脾、小肠、肾脏和肿瘤中可检测到绿色荧光蛋白的表达和相应细胞因子基因的整合。血清中相应的细胞因子水平较对照组明显升高 (P <0 .0 5 ) ,生存期远远超过对照组小鼠 (P <0 .0 5 )。结论 减毒沙门菌可作为口服基因治疗载体 ,可能为肿瘤的治疗提供一条简便、安全、有效的途径
Objective To explore the feasibility of using attenuated Salmonella as oral gene therapy vector. Methods The eukaryotic expression vectors pCMVhIL 12, pCMVhGM CSF and EGFPN1 were transfected into attenuated Salmonella typhimurium SL32 6 1 by electroporation and fed to BALB c and C5 7BL 6 mice via gastric tube. After 6 weeks, they were challenged with 4T1 breast cancer cells and Lewis lung cancer cells, respectively. The expression of green fluorescent protein (GFP) was detected by flow cytometry and confocal microscopy in each mouse. The integration and expression of IL-12 and GM-CSF genes were detected by PCR and ELISA. And investigated tumor suppression and mouse survival. Results The expression of green fluorescent protein and the integration of the corresponding cytokine genes were detectable in the liver, spleen, small intestine, kidney and tumor of mice. Serum corresponding cytokines levels were significantly higher than the control group (P <0. 05), the survival time is much longer than the control group mice (P <0. 05). Conclusion Attenuated Salmonella can be used as oral gene therapy vector, which may provide a simple, safe and effective way for the treatment of cancer