摘要
目的:研究hPSMA和4-1BBL基因转染小鼠树突状细胞(DC)后的变化及其在前列腺癌方面的临床意义。方法:以含hPSMA和4-1BBL基因的重组腺病毒载体转染C57BL/6j小鼠骨髓来源的DC,将培养的DC分为阳性组(Ad/hPSMA-IRES-m4-1BBL组)、阴性对照组(Ad5LacZ组)和未转染组,分别加入重组腺病毒、空载质粒腺病毒和生理盐水后,采用流式细胞仪测定腺病毒的转染效率;用相差显微镜、扫描电镜和透视电镜观察DC的形态学变化;用流式细胞仪分析表型变化;用Western blot检测DC hPSMA蛋白的表达。结果:经体外诱导培养后获得的树突状细胞在相差显微镜、透视电镜和扫描电镜下均观察到具有典型形态的成熟DC;流式细胞仪检测腺病毒转染效率为70%,Ad/hPSMA-IRES-m4-1BBL组DC细胞表面MHC-Ⅱ、CD80、CD86较其余两组无明显差异(P>0.05),但4-1BBL有明显提高(P<0.05);Western blot检测DC蛋白表达出PSMA。结论:本实验中获得的DC具有成熟DC的典型特征,DC表面的4-1BBL表达率明显上调,表达出PSMA蛋白。本研究将为DC应用于前列腺癌的生物治疗奠定基础。
Objective：To investigate the changes of the dendritic cell （DC） which was transfected by hPSMA and 4-1BBL genes as well as their clinic significance in prostate cancer. Methods： Using the recombinant adenovirus vector containing hPSMA and 4-1BBL genes to transfect the dendritic cells which were from C57BL/6j mouse bone marrow. Allocating the dendritic cells for three groups： positive group（Ad/hPSMA-IRES-m4- 1BBL group）, negative group （AdSLacZ group） and in transfected group, we added the recombinant adenovirus, the empty vector of adenovirus and physiological saline in groups respectively. And then detected the efficiency of adenovirus transfection efficiency by FACS and observing the morphologic changes of the dendritic cells under the phase contrast microscope, the scanning transmission electron micrographs and the transmission electron micrographs; the surface molecular expression of DCs were detected by FACS; At last, detection of expression of protein of hPSMA of DCs by Western blot. Results： The induced DCs in vitro displayed the typical morphological characteristics of mature DCs under the phase contrast microscope, the scanning transmission and the transmission electron micrographs. 70% of adenovirus transfected efficiency was reported by FACS. It is no obvious difference between the group of Ad/hPSMA-IRES-m4-1BBL and the other two groups（P〉0.05）in MHC-II , CD80 and CD86 of the surface of DCs. But the 4-1BBL which was one of the surface of DCs was increased（P〈0.05）. Conclusions： The obtained DCs displayed the typical morphological characteristics of mature DC, the rate of 4-1BBL expression of surface DC was obviously up-regulated and DCs expressed the protein of PSMA. This study would build theoretic basis for DIE biological treatment of prostate cancer.
出处
《临床泌尿外科杂志》
北大核心
2010年第3期230-234,共5页
Journal of Clinical Urology
基金
国家自然科学基金(编号30672107)
