与转染Endostatin基因的角朊细胞共培养时内皮细胞增殖特性变化

Effects of Co-cultured with Endostatin Gene Transfected Keratinocytes on Proliferative Characteristics of the Human Dermal Microvascular Endothelial Cells

构建含Endostatin基因的腺病毒载体,将Endostatin基因导入培养的角朊细胞,并采用套皿法共培养角朊细胞与内皮细胞,测定培养液中Endostatin含量、内皮细胞增殖周期各时相比例、内皮细胞凋亡及细胞抑制率。结果表明转染Endostatin基因的角朊细胞可有效表达并分泌Endostatin,连续培养3天后,培养液中Endostatin含量可达226ng/ml;与转基因角朊细胞共培养的内皮细胞凋亡百分数与抑制率分别为(32.7±7.1)%、(60.5±8.3)%,均显著高于对照组[(7.3±2.0)%,(13.8±1.6)%],且G0/G1期比例明显高于对照组,而S期、G2/M期比例及增殖指数显著低于对照组。因此,转染Endostatin基因角朊细胞与内皮细胞共培养时,角朊细胞可通过分泌Endostatin促进内皮细胞凋亡,并抑制其增殖。

Constructed a recombinant endostatin adenovirus expression vectors and transformed these vectors into keratinocytes, co-cultured the kerotinocytes with human dermal microvascular endothelial cells, then determined the content of endostatin in the supernate, the phase percent of the proliferative cycle, apoptosis and the cell inhibition ratio of the endothelial cells. The endostatin expression by keratinocytes reached 226 ng/ml after 3 days of co-culture. The apoptosis percentage and the cell inhibition ratio of the endothelial cells co-cultured with gene-tranfected keratinocytes were significantly higher than those in control group. In addition, the proportion of G0/G1 period was higher than that in control group, while the the proportion of S and G2/M period were on the other way round. Therefore, co-cultured gene-transfected keratinocytes could promote apoptosis and inhibite the proliferation of the endothelial cells through excretion of endostatin.