Sequences encoding PF4 (58-70) and TSP1 (429-459) were linked to yield a single gene TSF which encodes the fuse-protein of TSF. The gene was cloned into a pGEX-2T expression vector to generate a protein GST-TSF, which...Sequences encoding PF4 (58-70) and TSP1 (429-459) were linked to yield a single gene TSF which encodes the fuse-protein of TSF. The gene was cloned into a pGEX-2T expression vector to generate a protein GST-TSF, which was strongly expressed in E. coli. The purified GST-TSF was degraded with thrombin to generate the protein TSF. With the methods of MTT and wound repair assay, the effects of TSF on the proliferation and migration of EC were detected, respectively. The results showed that TSF significantly suppressed BAEC proliferation and migration in a dose-dependent manner. The fuse protein GST-TSF, and the peptides PF4 (58-70) and TSP1 (429-459) also inhibited BAEC proliferation and migration, respectively, but their inhibition rates were not as high as TSF. Using the CAM assay, it was shown that TSF, GST-TSF, PF4 (58-70) and TSP1 (429-459) inhibited angiogenesis in chick CAM potentially, the effect of TSF was the highest. In vivo, the growth of Lewis lung carcinoma was potently inhibited by TSF展开更多
A live attenuated AraA autotrophic mutant of Salmonella typhimurium (SL3261) was used as carrier for eukaryotic expression vectors EGFPN1, pCMVmIL-12, pCMVWL-12, pCMVmGM-CSF and PCMVhGM-CSF and was administered orally...A live attenuated AraA autotrophic mutant of Salmonella typhimurium (SL3261) was used as carrier for eukaryotic expression vectors EGFPN1, pCMVmIL-12, pCMVWL-12, pCMVmGM-CSF and PCMVhGM-CSF and was administered orally to BALB/c and C57BL/6 mice. After 6 weeks, these mice were challenged with 4T1 and Lewis tumor cells respectively. GFP expression and gene integration could be detected in mice’s livers, spleens, intestines, kidneys and tumors. The serum level of cytokines increasedsignificantly in treated mice, so did the ratio of CD8+/CD4+,which resulted in the tumor regression and prolongation of the survival time of those mice. These researches laid an experimental foundation for the tumor gene therapy using live attenuated salmonella.展开更多
基金This work was supported in part by the National Natural Science Foundation of China (Grant No. 39970814).
文摘Sequences encoding PF4 (58-70) and TSP1 (429-459) were linked to yield a single gene TSF which encodes the fuse-protein of TSF. The gene was cloned into a pGEX-2T expression vector to generate a protein GST-TSF, which was strongly expressed in E. coli. The purified GST-TSF was degraded with thrombin to generate the protein TSF. With the methods of MTT and wound repair assay, the effects of TSF on the proliferation and migration of EC were detected, respectively. The results showed that TSF significantly suppressed BAEC proliferation and migration in a dose-dependent manner. The fuse protein GST-TSF, and the peptides PF4 (58-70) and TSP1 (429-459) also inhibited BAEC proliferation and migration, respectively, but their inhibition rates were not as high as TSF. Using the CAM assay, it was shown that TSF, GST-TSF, PF4 (58-70) and TSP1 (429-459) inhibited angiogenesis in chick CAM potentially, the effect of TSF was the highest. In vivo, the growth of Lewis lung carcinoma was potently inhibited by TSF
文摘A live attenuated AraA autotrophic mutant of Salmonella typhimurium (SL3261) was used as carrier for eukaryotic expression vectors EGFPN1, pCMVmIL-12, pCMVWL-12, pCMVmGM-CSF and PCMVhGM-CSF and was administered orally to BALB/c and C57BL/6 mice. After 6 weeks, these mice were challenged with 4T1 and Lewis tumor cells respectively. GFP expression and gene integration could be detected in mice’s livers, spleens, intestines, kidneys and tumors. The serum level of cytokines increasedsignificantly in treated mice, so did the ratio of CD8+/CD4+,which resulted in the tumor regression and prolongation of the survival time of those mice. These researches laid an experimental foundation for the tumor gene therapy using live attenuated salmonella.
