中文摘要：

目的：分析B16-F10黑色素移植瘤及其肺转移瘤的差异表达蛋白，以筛选黑色素瘤转移相关的分子标志。方法：应用荧光差异凝胶电泳（two—dimensional differential gel electrophoresis，2D—DIGE）结合基质辅助激光解析电离飞行时间质谱技术matrix assisted laser desorption ionisation time—of-flight mass spectrometry，MALDI—TOF—MS）分离鉴定B16-F10黑色素移植瘤及其肺转移瘤的差异表达蛋白，部分差异蛋白经Real—time PCR进行mRNA表达水平验证。结果：Decyder6．O软件分析结果显示2D—DIGE图谱分辨率高、重复性好，30个蛋白点在实验组和对照组间存在表达差异（IRatiol≥2，P〈0．01），经质谱分析和数据库查询鉴定出9个蛋白在实验组表达上调，包括肌红蛋白（myoglobin，MB）、波形蛋白（vimentin，VIM）、磷酸甘油激酶1（phosphoglycerate kinase1，PGK1）、磷酸丙糖异构酶（Triosephosphate isomerase，TPI或TIM）、重链结合蛋白（heavy—chain binding protein，BiP）、α-烯醇化酶（α—enolase或enolasel）、β-肌动蛋白（β-actin）、γ-肌动蛋白（γ-actin）、层连蛋白结合蛋白（laminin—binding protein），这些蛋白主要参与了细胞骨架构成、糖酵解等生物学过程。Real—time PCR结果显示糖酵解酶PGK1及TPI mRNA表达水平在实验组显著高于对照组（P=0．001，0．003），变化趋势与蛋白质组学相一致。结论：小鼠黑色素瘤转移过程与多种蛋白的异常表达有关，糖酵解酶PGK1、TPI可能参与了黑色素瘤的转移过程。

英文摘要：

Objective： To investigate differentially expressed protein profiles in B16-F10 grafted melanoma and its metastasis in the lung in order to identify molecular markers of melanoma metastasis. Methods： Differentially expressed proteins in B16-F10 grafted melanoma and its metastatic lesion in the lung were isolated and identified by fluorescence two-dimensional differential gel electrophoresis （2D-DIGE） coupled with matrix assisted laser desorption ionisation time-of-flight mass spectrometry （MALDI-TOF-MS）. Some of identified proteins were further confirmed by Real-time PCR analysis. Results： High resolutional images of differential gel electrophoresis were obtained and 9 of 30 differentially expressed proteins （IRatiol ≥2, P〈0.01） were identified by MALDI-TOF-MS. The expression of Myoglobin （MB）, vimentin （VIM）, phosphoglycerate kinase 1 （PGK1）, Triosephosphate isomerase （TPI or TIM）, heavy-chain binding protein （BiP）, α-enolase, β-actin, γ-actin, and laminin-binding protein were up-regulated in the experimental group compared with the control group. These proteins were involved in the cytoskeletal formation, glycolysis and so on. Real-time PCR analysis showed up-regulation of mRNA expression of PGK1 and TPI in the experimental group （P=0.001 and 0.003）, which was in consistent with the results of proteomic analysis. Conclusion： A variety of abnormally expressed proteins contribute to the metastasis of mice melanoma. Glycolytic enzymes PGK1 and TPI may be involved in this process.