抗菌肽abps1真核表达载体的构建及其在肿瘤基因治疗中的初步应用-生物化学与分子生物学专业毕业论文.docxVIP

AbstractAntibacterial Abstract Antibacterial peptide(ABP)is a kind of small peptide which has a wide range of biological activities such as antimicrobial activities，antivirus and antitumor activities，To explore the antitumor activity of ABPSl which formerly designed according to the sequences of Shiva-1，firstly,a eukaryotic expression vector pVAXl一ABPSl was constructed and then the effect of it,when it was applied as a gane drug，on Rllnor cells and artificial cancer in nude mice was studied． The vector pVAXI-ABPSl was transfeeted into the tumor ceils HeLa and U25l by the lipofectamin-mediated DNA transfection procedure．6 hours post transfection，the morphological changes under light microscope and electronic microscope in HeLa cells could be observed significantly, In contrast with the control cells．the mierovilli彻the surface ofthe celIs whieh wee Iransfected with pVAXl-ABPSl were disrupted，decreased and even to disappeared．The cell membrane was injured and intracellular substances were leaked out．And finally the cell died．The same phenomena in U25 1 cells were also appeared in 12 hours aRer transfection．With the detection ofFlow Cytometric technique．the apeptosis COlVe was not found in HeLa and U25 1 cells changed in morphologically post transfeetion． A preliminary study of effect on tumor in vivo was carried out．The tunlor models in nude mice were established by subcutaneous injection ofHeLa cells in dorsum or former armpit． The injection of 10 u g pVAXl-ABPSl packed by lipofectamin into each tumor body was performed one time every day．The animals wel+e inspected and the tumor sizes were evaluated once in two days．After 7 days，they were killed．Every tumor bodies were carefully weighed while removed from nude mouse body by blund dissection These results gained in vitro and in vivo research suggested that the delivery of ABPSlgene into hunor cells or artificial cancelwould be effective in Carlcer gene therapy． Key words：ABPSl，tumor，nude mice，gene therapy II 独创性声明本人声明