皮肤角朊细胞与表皮干细胞双向电泳图谱的建立及差异分析

目的 建立皮肤角朊细胞与表皮干细胞的双向电泳图谱,并分析二者表达蛋白质的差异,为进一步研究体外调控表皮十细胞的增殖、分化提供线索.方法酶消化法获取单个表皮细胞悬液.Ⅳ型胶原快速贴壁法分选表皮干细胞.利用舣向电泳技术(2-DE)建立两种细胞的蛋白质表达图谱,并用Imagemaster 2D elite 5.0软件分析两种细胞的差异表达蛋一点.结果两种细胞的2-DE蛋白表达谱具有良好的重复性和可比性.皮肤角朊细胞与表皮干细胞的电泳图谱平均蛋白质点数分别为(982 ±18)个和(930 ±15)个,匹配点数为(850±13)个和(798±11)个,匹配率是86.56%和85.81%;两种细胞蛋白质间匹配点数是(886 ±8)个,匹配率是76.98%.找到差异蛋白点11个,其中只在表皮下细胞中表达或高表达的有8个;只在皮肤角朊细胞中表达或高表达的有3个.结论利用双向电泳法得到了分辨率较高儿重复性好的皮肤角朊细胞与表皮干细胞蛋白质组图谱,且二者存在有一定的差异.

Construction and difference analysis of two-dimensional gel electrophoresis profiles about keratinocytes and epidermal stem cells

Objective To establish the two-dimensional gel electrophoresis (2-DE) map of keratinocytes and epidermal stem cells, to analyze the differentially expressed proteins between two type cells and provide clues for regulating the proliferation and differentiation of epidermal stem cells in vitro. Methods The single epidermal cell suspension was obtained by enzyme digestion and the epidermal stem cells were harvested based on collagen type Ⅳ rapidly adhesiveness. The protein expression maps of the two types of cells were obtained using 2-DE, and the differential expressed protein spots were analyzed by linage master 2D elite 5.0 software. Results There were good reproducibility and comparability between two cell lines from the 2-DE protein expression maps. Average protein spots were 982 ±18 in keratinocytes and 930 ± 15 in epidermal stem cells. The matched spots were 850 ± 13 and 798 ±11 in these two types of cells, and the average matching rates were 86.56% and 85.81%. There were 886 ± 8 matching protein spots between two types of cells, and the average matching rate was 76.98%. Eleven differential protein spots were identified between two types of cells. Eight protein spots were expressed only or higher in the epidermal stem cells and three protein spots were expressed only or higher in the keratinocytes. Conclusion The proteome maps of keratinocytes and epidermal stem cells with high resolution and reproducibility can be obtained by the technique of 2-DE and some differences exist in protein expression between these two types of cells.