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Vegfr2-luc转基因子代小鼠的鉴定

ISSN号：1009-3419
期刊名称：《中国肺癌杂志》
时间：0
分类：R734.2[医药卫生—肿瘤;医药卫生—临床医学]
作者机构：[1]天津医科大学总医院、天津市肺癌研究所、天津市肺癌转移与肿瘤微环境实验室,天津300052, [2]天津医科大学总医院胸部肿瘤中心,天津300052
相关基金：国家自然科学基金重点项目(No.30430300);“973”重点项目(No.2010CB529405);天津市科技支撑计划重大项目(No.07SYSYSF05000)和重点项目(No.06YFSZSF05300)资助~~

作者：王伟强[1], 刘红雨[1], 宋志涛[1], 王玉丽[1], 王竟[1], 李颖[1], 李永文[1], 王岷[1], 陈军[1,2], 周清华[1,2]

关键词：动物模型, 血管新生, 荧光素酶报告基因, Animal model, Angiogenesis, Luciferase

中文摘要：

背景与目的 Vegfr2-luc转基因小鼠体内血管内皮生长因子受体2（vascular endothelial growth factor recep- tor 2,VEGFR2）的表达可以驱动荧光素酶报告基因（luciferase,luc）的表达,是活体动物水平实时监测血管生成情况的有利工具。本研究旨在对子代Vegfr2-luc转基因小鼠进行鉴定,以确定能否用于血管生成研究。方法 PCR检测新生小鼠基因组内luc基因;利用活体成像技术观察新生Vegfr2-luc转基因小鼠生长发育过程中以及皮肤伤口修复过程中 luc基因表达水平的变化情况;荧光素酶报告基因检测试剂盒检测成年（8周龄）转基因小鼠各脏器荧光素酶的活性和Real-time PCR检测各器官VEGFR2 mRNA的表达水平。结果 PCR结果显示50%（56/112）的新生小鼠携带luc基因。活体成像结果显示随着Vegfr2-luc转基因小鼠发育成熟,luc表达量逐渐降低（P〈0.001）;在皮肤伤口修复过程中, 伤口处luc表达水平先增强后降低（P〈0.001）。雌性成年转基因小鼠各脏器VEGFR2 mRNA的表达水平与荧光素酶活性呈正相关（r=0.948,P〈0.001）。将睾丸组织除外,雄性成年转基因小鼠各脏器VEGFR2mRNA的表达水平与荧光素酶活性同样呈正相关（r=0.836,P〈0.001）。结论 Vegfr2-luc转基因子代小鼠体内luc表达水平的变化可以反映VEGFR2 的表达情况。

英文摘要：

Background and objective A transgenic mouse,Vegfr2-luc,in which a luciferase reporter（luc） is under control of the murine VEGFR2 promoter,can be used to track angiogenesis in vivo. The aim of this study is to identify the offspring of Vegfr2-luc transgenic mouse. Methods Luc was detected with PCR in genomic DNA of the new-born mouse. Luc expression in the offspring of Vegfr2-luc transgenic mouse was monitored with IVIS in vivo imaging system during post-natal development. Wound-healing models of Vegfr2-luc transgenic mouse offspring were established and the expression of luc was monitored during the wound-healing process. Luc activity and VEGFR2 mRNA expression in different organs were detected with luc Assay System and Real-time PCR respectively. Results PCR showed that 50%（56/112） of the offspring of Vegfr2-luc transgenic mouse carry luc. IVIS in vivo imaging results demonstrated that luc expression in Vegfr2-luc transgenic mouse dropped dramatically with age increase（P0.001） and luc expression in the wound first increased and then decreased during the wound-healing process（P0.001） . Luc activity in female Vegfr2-luc transgenic mouse organs was positively correlated with VEGFR2 mRNA expression（r=0.948,P0.001） . Except testis,luc activity in male Vegfr2-luc transgenic mouse organs was also positively correlated with VEGFR2 mRNA expression（r=0.836,P0.001） . Conclusion The offspring of Vegfr2-luc transgenic mouse is applicable to tracking angiogenesis in vivo.

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