[Development of a novel protein carrier inducing immune response and binding DNA in gene therapy].
OBJECTIVE
To develop a novel protein carrier which can not only regulate the immune system but also deliver DNA into the tumor cell as an effective non-viral gene delivery system.


METHODS
By using gene engineering techniques, we constructed a fusion protein containing the -COOH end of human hepatitis B virus core antigen (HBcAg), small home-to-cancer peptide ligand RGD and Glutathione S-transferase (GST), which was expressed in E. coli and purified by size exclusion chromatography and affinity chromatography. We labeled it with FITC to observe whether it could bind prostate cancer PC-3 cell lines, and meanwhile used it as a non-viral gene delivery carrier with the plasmid pEGFP-N1 that could express GFP in PC-3 cells. Furthermore, we observed the regulatory function of this fusion protein to the mouse immune system.


RESULTS
The results of SDS-PAGE showed that the new protein carrier was obtained, which It could enter PC-3 cells with DNA in vitro and induce the mouse immune system to produce IgG1 and IgG2alpha simultaneously.


CONCLUSIONS
The new protein carrier can be used as a target protein, especially in positive cells and the immune system. It promises to be a good novel carrier for the gene therapy of cancer.
Shang
Xue-jun
XJ
Department of Andrology, School of Clinical Medicine, Nanjing University Medical College, Nanjing General Hospital of Nanjing Military Region, Nanjing, Jiangsu 210002, China. shangxj@androl.cn
Ge
Jing-ping
JP
Huang
Wei-dong
WD
Yao
Gen-hong
GH
Li
Hong-jun
HJ
Huang
Yu-feng
YF
D000818:Animals; D004247:DNA / Q000235:genetics* / Q000276:immunology*; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics* / Q000276:immunology*; D005980:Glutathione Reductase / Q000235:genetics; D006512:Hepatitis B Core Antigens / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D009842:Oligopeptides; D010957:Plasmids; D011994:Recombinant Proteins
D006512:Hepatitis B Core Antigens; D009842:Oligopeptides; D011994:Recombinant Proteins; C047981:arginyl-glycyl-aspartic acid; D004247:DNA; D005980:Glutathione Reductase