Skoda R C, Tsai S F, Orkin S H, Leder P
Howard Hughes Medical Institute, Department of Genetics, Harvard Medical School, Boston, Massachusetts, USA.
J Exp Med. 1995 May 1;181(5):1603-13. doi: 10.1084/jem.181.5.1603.
To study oncogenesis in the erythroid lineage, we have generated transgenic mice carrying the human c-MYC proto-oncogene under the control of mouse GATA-1 regulatory sequences. Six transgenic lines expressed the transgene and displayed a clear oncogenic phenotype. Of these, five developed an early onset, rapidly progressive erythroleukemia that resulted in death of the founder animals 30-50 d after birth. Transgenic progeny of the sixth founder, while also expressing the transgene, remained asymptomatic for more than 8 mo, whereupon members of this line began to develop late onset erythroleukemia. The primary leukemic cells were transplantable into nude mice and syngeneic hosts. Cell lines were established from five of the six leukemic animals and these lines, designated erythroleukemia/c-MYC (EMY), displayed proerythroblast morphology and expressed markers characteristic of the erythroid lineage, including the erythropoietin receptor and beta-globin. Moreover, they also manifested a limited potential to differentiate in response to erythropoietin. Studies in the surviving transgenic line indicated that, contrary to our expectations, the transgene was not expressed in the mast cell lineage. That, coupled with the exclusive occurrence of erythroleukemia in all the transgenic lines, suggests that the GATA-1 promoter construct we have used includes regulatory sequences necessary for in vivo erythroid expression only. Additional sequences would appear to be required for expression in mast cells. Further, our results show that c-MYC can efficiently transform erythroid precursors if expressed at a vulnerable stage of their development.
为了研究红系细胞系中的肿瘤发生,我们构建了在小鼠GATA-1调控序列控制下携带人c-MYC原癌基因的转基因小鼠。六个转基因品系表达了转基因并表现出明显的致癌表型。其中,五个品系发生了早发性、快速进展的红白血病,导致奠基动物在出生后30-50天死亡。第六个奠基者的转基因后代虽然也表达转基因,但在超过8个月的时间里没有症状,此后该品系的成员开始发生迟发性红白血病。原发性白血病细胞可移植到裸鼠和同基因宿主中。从六只白血病动物中的五只建立了细胞系,这些细胞系被命名为红白血病/c-MYC(EMY),表现出早幼红细胞形态,并表达红系细胞系特有的标志物,包括促红细胞生成素受体和β-珠蛋白。此外,它们对促红细胞生成素的反应也表现出有限的分化潜能。对存活的转基因品系的研究表明,与我们的预期相反,转基因在肥大细胞系中不表达。这一点,再加上所有转基因品系中都只发生红白血病,表明我们使用的GATA-1启动子构建体仅包含体内红系表达所需的调控序列。在肥大细胞中表达似乎还需要其他序列。此外,我们的结果表明,如果c-MYC在红系前体细胞发育的易损阶段表达,它可以有效地转化这些细胞。