OSM对黑色素瘤的抑制作用与基因-放射治疗效果

目的：通过细菌内同源重组，制备含绿荧光（GFP）基因的CD腺病毒，初步观察其体外杀瘤作用。方法：先构建含CD基因的腺病毒梭载体pAdTrack-CMV-CD,与骨架载体pAdEasy-1在细菌内重组为pAd-CD，经293细胞包装为增殖缺陷性腺病毒，体外感染人结肠癌细胞株LoVo，并给予前药5－FC，观察其体外杀瘤效果。结果：pAdTrack-CMV-CD和同源重组的pAd-CD经酶切鉴定正确，pAd-CD转染293细胞，包装扩散腺病毒后，感染LoVo细胞，予5－FC治疗，4d后在荧光温微镜下观察到转染LoVo和未转染细胞胞均被杀死，并且 这种旁观者效应不依赖于细胞间接解。结论：细菌内质粒同源重组法可以高效制备含绿荧光（GFP)基因的CD腺病毒，体外腺病毒介导CD／5－FC可以杀伤肿瘤细胞，其旁观者效应不依赖于细胞间接触。

Oncostatin M ShowsInhibitory Role and Effect of Gene-Radiotherapy on Melanoma Growth in vivo

Objective: To generate an adenovirus, Ad-CD, expressing green fluorescence protein and bacterial cytosine deaminase, and use it for tumor suicide gene therapy. Methods: CD gene was cloned into pAdTrack-CMV, pAdTrack-CMV-CD and pAdEasy-1 were homologous recominated in bacteria. 293 cells were transfected by the newly-recombinated plasmid pAd-CD and generated adenoviruses. LoVo cells were infected with the adenovirus, and the tumor-killing effect after 5-FC was administered.Results: pAdTrack-CMV-CD and the newly-recombinated plasmid pAd-CD were tested by restriction endonuclease digestions. Followed by GFP, Ad-CD were produced in 293 and LoVo cells infected by Ad-CD were killed after 5-FC was administered.Conclusion: Adenovirus plasmid homologous recombinant in bacteria is a simple method for adenovirus generation. In vitro adenoviral carried CD gene is efficient for tumor cell suicide gene therapy and has a bystand-effect, which is not depended on cell direct contact