腺病毒介导的KGF基因修饰促进表皮干细胞增殖

目的　构建KGF腺病毒表达载体并转染表皮干细胞，观察其对表皮干细胞增殖的影响。 方法　 RT-PCR自人成纤维细胞中扩增KGF基因片段，pAD-easy I腺病毒系统构建KGF腺病毒表达载体，以MOI=50感染表皮干细胞；MTT法检测表皮干细胞增殖，流式分析细胞周期变化。 结果　　RT-PCR成功扩增KGF基因片段，成功构建的KGF腺病毒质粒经Pac I内切酶酶切获得4.5 kbp的特异条带，MOI=50的病毒转染效率超过90%；KGF基因修饰后的表皮干细胞增殖效率与对照组比较有显著差异（P<0.05），并且G2/M期细胞数量明显增多。 结论　成功构建腺病毒KGF表达载体，腺病毒载体介导的KGF基因修饰促进了表皮干细胞的增殖。

Effect of recombinant adenovirus vector carrying human KGF gene on the proliferation of human epidermal

Objective To construct a recombinant adenovirus vector carrying human keratinocyte growth factor(KGF) gene,and investigate its proliferative effect on the transferred primary human epidermal stem cells.Methods RT-PCR was used to amplify the sepuence of human keratinocyte growth factor.Recombinant adenoviral vector containing the sequence was prepared using pAD-easy-I system.The primary human epidermal stem cells were transfected with the recombinant virus vector.Under the fluorescence microscope,the transfe...