摘要
探讨减毒鼠伤寒沙门氏菌作为口服基因治疗载体的可行性 .通过电转化法将真核表达载体 pCMVmIL 12、pCMVmGM CSF、EGFPN1导入减毒鼠伤寒沙门氏菌SL32 6 1中 ,经由胃管饲于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的方法检测mIL 12、mGM 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 .结果表明 :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 ) .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。
To study the possibility of oral gene therapy using live attenuated Salmonella. A live attenuated AraA(-) autotrophic mutant of Salmonella typhimurium (SL3261) was used as carrier for eukaryotic expression vectors EGFPN1, pCMVmIL-1,2, pCMVmGM-CSF and was administered orally to BALB/c and C57BL/6 mice. After 6 weeks, these mice were challenged with 4T(1) or Lewis tumor cells respectively. Flow cytometer and laser scanning confocal microscopy were used to detect the expression of GFP in murine tissues. PCR and ELISA were used to detect the integration and expression of mIL-12, mGM-CSF gene. The survival time of mice was also investigated. GFP expression and mIL-12, mGM-CSF gene integration could be detected in murine liver, spleen, intestine, kidney and tumor. The serum level of mIFN-gamma, mIL-12 increased significantly in the mIL-12 orally treated mice (P < 0.05); The serum level of mGM-CSF increased also in mGM-CSF orally treated mice (P < 0.05); which resulted in the prolongation of the survival time of those mice, compared with the control (P < 0.05). Oral gene therapy using live attenuated Salmonella has the potential to be a simple, effective and above all, safe way against tumor.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2001年第4期537-541,共5页
Progress In Biochemistry and Biophysics
