Heat shock-inducible Cre/Lox approaches to induce diverse types of tumors and hyperplasia in transgenic zebrafish
Heat shock-inducible Cre/Lox approaches to induce diverse types of tumors and hyperplasia in transgenic zebrafish
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DOI：
10.1073/pnas.0611302104
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发表时间：
2007-05-29
影响因子：
11.1
通讯作者：
Zon, Leonard I.
中科院分区：
文献类型：
--
作者：
Le, Xiuning;Langenau, David M.;Zon, Leonard I.
RAS家族成员是人类癌症中最常突变的癌基因之一。鉴于斑马鱼在化学和遗传筛选中的实用性，开发RAS诱导的癌症模型将使大规模筛选成为可能，以进一步了解恶性肿瘤的分子机制。我们开发了一种热休克诱导的Cre/Lox介导的转基因方法，其中通过热休克处理可以在转基因动物中有条件地诱导激活的人kRASG 12 D。具体而言，双转基因鱼Tg（B-actin-LoxP-EGFP-LoxP-kRASG 12 D，hsp 70-Cre）在整个斑马鱼胚胎热休克后发生了四种类型的肿瘤和增生，包括横纹肌肉瘤、骨髓增生性疾病、肠道增生和恶性外周神经鞘瘤。使用离体热休克和移植来自双转基因动物的全肾骨髓细胞，我们能够在受体鱼中特异性地产生kRASG 12 D诱导的骨髓增生性疾病。这种热休克诱导的重组方法允许产生多种类型的RAS诱导的肿瘤和增生，而不表征组织特异性启动子。此外，这些肿瘤和增生在形态学和分子水平上与人类疾病非常相似。
RAS family members are among the most frequently mutated oncogenes in human cancers. Given the utility of zebrafish in both chemical and genetic screens, developing RAS-induced cancer models will make large-scale screens possible to understand further the molecular mechanisms underlying malignancy. We developed a heat shock-inducible Cre/Lox-mediated transgenic approach in which activated human kRASG12D can be conditionally induced within transgenic animals by heat shock treatment. Specifically, double transgenic fish Tg(B-actin-LoxP-EGFP-LoxP-kRASG12D, hsp70-Cre) developed four types of tumors and hyperplasia after heat shock of whole zebrafish embryos, including rhabdomyosarcoma, myeloproliferative disorder, intestinal hyperplasia, and malignant peripheral nerve sheath tumor. Using ex vivo heat shock and transplantation of whole kidney marrow cells from double transgenic animals, we were able to generate specifically kRASG12D-induced myeloproliferative disorder in recipient fish. This heat shock-inducible recombination approach allowed for the generation of multiple types of RAS-induced tumors and hyperplasia without characterizing tissue-specific promoters. Moreover, these tumors and hyperplasia closely resemble human diseases at both the morphologic and molecular levels.