six1基因对黑素瘤发生、发展及转移的影响初探word论文.docxVIP

目录英文缩略词表...............................................................................................1中文摘要.......................................................................................................2Abstract3引言...........................................................................................................5材料和方法...................................................................................................7结果.........................................................................................................18讨论.........................................................................................................24参考文献.....................................................................................................27综述.........................................................................................................31参考文献.....................................................................................................38附录：硕士研究生阶段参加会议情况及发表论文.................................45致谢.........................................................................................................46英文缩略词表缩写英文全称中文全称MM CMMSix1 VEGF-C VEGFR-3shRNA IHC cDNA RT-PCR WB TGF-β MAPKMalignant melanoma cutaneous malignant melanoma sine oculis homeoboxhomologVascular endothelialgrowth factor-C Vascular endothelialgrowth factor receptor-3short hairpin RNA Immunohistochemistry complementaryDNA real timeRNA Western blotTransforming growth factor-beta Mitogen-actived proteinkinase恶性黑素瘤皮肤恶性黑素瘤同源异型基因或蛋白six1 血管内皮生长因子C 血管内皮生长因子受体3短发夹RNA 免疫组织化学互补DNA半定量聚合酶链反应免疫印迹转化生长因子β促分裂原活化蛋白激酶six1 基因对黑素瘤发生、发展及转移的影响初探硕士研究生：杨珍导师：陈思远教授华中科技大学同济医学院附属协和医院皮肤科中文摘要[目的]研究小鼠黑素瘤B16F10 细胞中six1、VEGF-C mRNA及蛋白质的表达情况，探讨six1 基因在小鼠黑素瘤发生、发展及转移中所发挥的作用。[方法]针对six1 基因序列设计并合成短发夹RNA（six1-shRNA）质粒载体，转染入小鼠B16F10细胞，筛选出稳定干扰细胞株。用Realtime-PCR和Western Blot 分别检测细胞six1和VEGF-C 的表达情况；并将细胞株注入C57BL/6小鼠皮下成瘤，观察小鼠肿瘤生长、转移情况，并用HE染色及免疫组织化学法检测瘤组织six1、VEGF-C 表达及VEGFR-3标记的淋巴管情况。[结果]①six1-shRNA质粒B16F10细胞转染率20-30%。②稳定转染si