摘要
目的探讨以减毒鼠伤寒沙门氏菌为载体的口服细胞因子基因治疗防治小鼠肿瘤的可行性。方法通过电转化法将真核表达载体pCMVmIL-12、EGFPN1导入减毒鼠伤寒沙门氏菌SL3261中,经由胃管饲于BALB/c和C57BL/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方法检测mIL-12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升高(P<0.05),生存期超过对照组小鼠(P<0.05)。结论减毒沙门氏菌可作为口服基因治疗载体,为肿瘤的预防和治疗提供一条简便、安全、有效的途径。
Objective To evaluate the feasibility of oral cytokine gene therapy against tumor using live attenuated Salmonella as a vector. Methods A live attenuated AraA- autotrophic mutant of Salmonella typhimurium （SL3261） was used as carrier for eukaryotic expression vectors EGFPN1 and pCMVmIL-12 administered orally in BALB/c and C57BL/6 mice. After 6 weeks, the mice were challenged with 4T1 or Lewis tumor cells, respectively, and flow cytometry and confocal microscopy were used to detect the expression of green fluorescence protein （GFP） in the tissues. PCR and ELISA were performed to detect the integration and expression of mIL-12 gene, and the survival time of the mice was also recorded. Results GFP expression and mIL-12 gene integration could be detected in the liver, spleen, intestinal, kidney and tumor tissues of the mice. The serum level ofmIFN-% mIL-12 increased significantly in mice with oral raiL-12 administration （P〈0.05）, which resulted in the survival prolongation of the mice as compared with the control mice （P〈0.05）. Conclusion Oral gene therapy using live attenuated Salmonella can be potentially a simple, effective and above all, safe means for tumor treatment.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2006年第12期1738-1741,共4页
Journal of Southern Medical University
基金
广东省自然科学基金博士科研启动基金(04300412)~~
