Preparation Of McAb Au_14-1-liposome-p53 Gene Carrier For Transfection Cervix Cancer And Expression In U₁₄ Cell Line

P53 gene ,one of tumor suppressor genes, which have intimate connection with the occurance and progression of many tumors in human, mainly induce the tumor cells apoptosis and cell period arrest. Using gene transfection techniques to transfer the wild type p53 cDNA into tumor cells can induce tumor cells apoptosis and inhibition of progression. Another important progression of gene transfection techniques is using specific monoclonal antibodies as carriers of transfection to take gene therapies of tumors through the targeted effect of McAb. This research assume to adopt the cationic liposomes as carriers, to transfect the U14 cell line of cervix cancer in vitro, to compare and observe the effect of McAb-CL-mediated transfection, and to provide the experimental according for exploring McAb-CL-mediated transfection in vivo and the gene therapies of cervix cancer.The experiment used the modified alkaline lysis method to extract and purify the wt-p53 plasmid DNA from amplified E. coli; used the caprylic acid/ammonium sulfate method to purify Au14-1 to conjugate with poly-lysine with positive charges and to prepare the conjugated complex; composed the PL(LipofectinR),plasmids DNA and Au14-1-PL conjugates to produce ternary complex by static electronic effects. The experiment was divided into seven groups: blank control, DNA, DNA+LP, Au14-1+DNA, Au14-1+DNA+LP, M5+DNA and M5+DNA+LP. M5 belongs to McAb (anti-parasite) for control. Add RPMI1640 completed culture medium containing 10% FCS to culture for 48hs continously twelve hours after transfectin instantly the cervix cancer U14 cells cultured in suspension, and then compare and observe the p53 expression through indirect cell enzyme immunol chemistral method.The experiment showed that the three groups of DNA-LP, Au14-1-DNA-LP and Au14-1-DNA all can be observed the specific stains of p53 nuclear proteins. The groups of bland control, DNA, M5-DNA-LP and M5-DNA, however, can not be seen the specific positive reactions. Furthermore, the diffrences between three positive groups cannot be observed through diluted experiment of antibody titres.The results of this experiment suggests that the target carrrier of gene transfection constructed by McAb Au14-1 can specifically mediate the transfer and expression of p53 gene, and the effection of expression excells the effecor of transfection by cationic liposome.