Bouchard L, Gelinas C, Asselin C, Bastin M
Virology. 1984 May;135(1):53-64. doi: 10.1016/0042-6822(84)90116-8.
A procedure has been developed whereby the oncogenicity of the DNA from polyoma (Py) virus and Simian virus 40 (SV40) can be tested directly by injecting recombinant DNA into newborn rodents. Injection of 0.2-2.0 micrograms of linear DNA induced the development of subcutaneous liposarcomas and fibrosarcomas at the site of inoculation. Coinjection of high-molecular-weight rat DNA as carrier had little or no effect on tumor formation but plasmids pBR322, pAT153 , and pML2 behaved as strong inhibitors. Tumor induction by injecting DNA into newborn rodents provides an in vivo equivalent to a transformation assay but appears to be a more stringent and rigorous criterion of oncogenic transformation. The oncogenic potential of Py virus in newborn hamsters could be expressed by a recombinant encoding only the middle T protein, although with average tumor latencies 5-10 times longer than those observed with wild-type Py DNA. Py middle T required the cooperation from small T to induce tumors in newborn rats. SV40 DNA was tumorigenic only in newborn hamsters. delta 2005 DNA which is unable to produce the SV40 small T antigen was much less active and required a latent period about twice that of wild-type SV40 DNA. However, its tumorigenic potential was restored by addition of the Py small T antigen gene. This indicates that Py and SV40 small T antigens are interchangeable and that they probably play an identical role in malignant transformation. Finally, evidence was provided that intermolecular recombination or recombination between DNA fragments can occur in vivo.
已开发出一种方法,通过将重组DNA注射到新生啮齿动物体内,可直接检测多瘤(Py)病毒和猴病毒40(SV40)的DNA的致癌性。注射0.2 - 2.0微克线性DNA可在接种部位诱发皮下脂肪肉瘤和纤维肉瘤。共注射高分子量大鼠DNA作为载体对肿瘤形成几乎没有影响,但质粒pBR322、pAT153和pML2表现为强抑制剂。通过向新生啮齿动物注射DNA诱导肿瘤,提供了一种体内相当于转化试验的方法,但似乎是致癌转化的更严格标准。Py病毒在新生仓鼠中的致癌潜力可由仅编码中间T蛋白的重组体来表达,尽管平均肿瘤潜伏期比野生型Py DNA观察到的长5 - 10倍。Py中间T需要小T的协同作用才能在新生大鼠中诱导肿瘤。SV40 DNA仅在新生仓鼠中具有致瘤性。无法产生SV40小T抗原的δ2005 DNA活性低得多,潜伏期约为野生型SV40 DNA的两倍。然而,通过添加Py小T抗原基因可恢复其致癌潜力。这表明Py和SV40小T抗原是可互换的,并且它们可能在恶性转化中发挥相同的作用。最后,有证据表明分子间重组或DNA片段之间的重组可在体内发生。