Wnt-1重组腺病毒的构建及其对人表皮干细胞分化的影响  

Construction of recombinant adenoviral vector carrying Wnt-1 and its influence on differentiation of epidermal stem cells

作　　者：李元朝[1] 伍津津[1] 张一[2] 杨涛[1] 杨桂红[1] 王兵强[1] 

机构地区：[1]第三军医大学大坪医院野战外科研究所皮肤科,重庆400042 [2]第三军医大学大坪医院野战外科研究所第一研究室

出　　处：《中华实验外科杂志》2008年第8期1053-1054,共2页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（30570972）

摘　　要：目的构建Wnt-1重组腺病毒，并观察Wnt-1对人表皮干细胞分化相关蛋白表达的影响。方法连接穿梭质粒与骨架质粒，构建重组腺病毒；扩增后感染人表皮干细胞，检测其3种角蛋白和基质金属蛋白酶（MMP）-2的表达。结果重组质粒酶切后可得到30kb和4．5kb两条特异性条带；人表皮干细胞感染重组腺病毒后，聚合酶链反应（PCR）与绿色荧光蛋白均指示Wnt-1基因的表达，且角蛋白10表达变为阳性；角蛋白18表达增强；角蛋白19表达减弱；细胞培养基中MMP-2浓度由（-16．25±2．40）μg/L升高至（714．88±30．45）μg/L（t=58．45，P〈0．01）。结论Wnt-1具有促使人表皮干细胞向腺样上皮细胞分化的趋势。Objective To construct recombinant adenovirus vector carrying Wnt-1 and observe its influence on differentiation of epidermal stem cells. Methods pAdTrack-CMV-Wnt-1 and pAdEasy-1 were linked to construct recombinant adenovirus vector. After the epidermal stem cells were infected with recombinant adenovirus,3 kinds of keratin and MMP-2 were detected. Results Recombinant adenovirus vector could be incised to 30 kb and 4.5 kb specificity straps. After epidermal stem cells were transfected with Wnt-1 gene, keratin10 expression became positive, keratin 18 expression was enhanced and keratin 19 expression reduced;Concentrations of MMP-2 were stepped up from （ -16.25 ± 2.40 ） μg/L to （714.88 ± 30.45） μg/L （t = 58.45, P 〈 0.01 ）. Conclusion Wnt-1 could induce differentiation of epidermal stem cells into glandular epithelium-like cells.

关 键 词：WNT-1 腺病毒载体 表皮干细胞 

分 类 号：R3[医药卫生—基础医学]