重组hFGF-10腺病毒对HaCat细胞影响的蛋白质组学研究

研究重组腺病毒导入人成纤维细胞生长因子10 (hFGF-10) 对HaCat细胞蛋白质组的影响, 由鉴定的差异蛋白质推测hFGF-10对HaCat细胞作用的可能机制。采用双向凝胶电泳结合串联飞行时间质谱技术, 对空载体腺病毒Ad感染细胞和重组腺病毒rAd-hFGF-10感染细胞的总蛋白图谱上的差异蛋白点进行鉴定, 并通过半定量RT-PCR和Western blotting在转录和翻译水平对差异蛋白进行确证。结果显示, 获得了蛋白质分离效果较好的双向凝胶电泳图谱, 鉴定了4种与细胞凋亡、细胞骨架调控、蛋白质降解等相关的差异蛋白质, 并在转录和翻译水平确证了差异蛋白质VDAC2在导入目的基因hFGF-10的HaCat细胞中表达上调, 提示VDAC2可能在hFGF-10的生物学功能中发挥作用。

Proteomic study of the effect of recombinant hFGF-10 adenovirus on HaCat cells

This study is to investigate the effect of recombinant hFGF-10 adenovirus on the proteome     of HaCat cells, and to speculate further the possible mechanism of the effect of hFGF-10 on HaCat cells via  differentially expressed proteins identified.  Two-dimensional gel electrophoresis (2-DE) combined with tandem time-of-flight mass spectrometry was applied to identify the differentially expressed protein spots on the 2-DE maps of the whole-cell proteins from Ad infected and rAd-hFGF-10 infected HaCat cells.  The mRNA and  protein levels of the differentially expressed proteins were confirmed with semi-quantitative RT-PCR and   Western blotting.  The results showed that the 2-DE maps with high resolution were obtained, and four selected differentially expressed proteins involved in cell apoptosis, cytoskeleton regulation and protein degradation  were identified with MALDI-TOF/TOF.  The mRNA and protein levels of one of the differentially expressed proteins, VDAC2, were up-regulated in HaCat cells infected with the recombinant hFGF-10 adenovirus.  The differentially expressed protein, VDAC2, may be related to the bioactivities of hFGF-10.