白血病相关的激活突变 SHP-2对小鼠体内血管新生能力的影响

摘要

Objective To study the effect of the activated mutant SHP-2 on murine angiogenesis and its role in tumorigenesis. Methods Heterozygous SHP-2 D61G mutant knock in and wild type C57BL/6 mice were used. The Matrigel containing FGF, VEGF was injected in the local subcutaneous of mouse to induced the angiogenesis. The method of Colorimetric Technique was used to detect the content of Hemoglobin (HB) in the matrigel to represent vascular density. The mouse peripheral blood HB concentration was assayed by the Blood Count Instrument for balance of HB level in matrigel. Then the matrigel was fixed and section was made and done Immunohistochemistry (IHC) for CD31 positive endothelial cell analysis. Finally the Transwell system was used to detect the migrating ability of mouse embryonic fibroblasts (MEFs). Result The HB level was higher in matrigel from SHP-2 D61G mutant mice than that from WT control mice. CD31 positive cells were increased dramatically in matrigel from SHP-2 mutant mouse than that from WT control group(P<0.01). The migration of SHP-2 D61G mutant MEFs were much more than WT MEFs (P<0.01). Conclusion Leukemia related mutant SHP-2 may promote mouse angiogenesis and the promotion effect may be related with the enhanced migrated ability of SHP-2 mutant cell.%目的 研究激活突变SHP-2对小鼠体内血管新生的影响,以进一步研究其在肿瘤发展过程中的作用.方法 利用SHP-2 D61G激活突变敲入(knock in)模型及WT(wild type)小鼠,局部皮下注射合成纤维细胞生长因子(FGF)、血管内皮生长因子(VEGF)的基质胶)(MatrigelTM Basement Membrane Matirx),诱导血管新生,用比色法检测基质胶内血红蛋白(HB)的含量代表血管密度,用血细胞计数仪检测相应小鼠外周血HB浓度用以标准化基质胶内HB含量;并用免疫病理学方法检测基质胶(Matrigel)内血管内皮细胞密度(CD31阳性);用Transwell方法检测小鼠胚胎成纤维细胞(mouse embryonic fibroblasts,MEFs)迁移能力.结果 SHP-2 D61G杂合突变的小鼠(SHP-2 D61G/+)体内注射的基质胶栓内HB含量明显高于WT组(P＜0.01),CD31阳性细胞也明显增多,SHP-2 D61G突变小鼠MEFs迁移能力明显增强(P＜0.01).结论 SHP-2 D61G激活突变可能促进小鼠体内血管新生,这种促进作用可能与细胞迁移能力增强有关.