Mackey J K, Rigden P M, Green M
Proc Natl Acad Sci U S A. 1976 Dec;73(12):4657-61. doi: 10.1073/pnas.73.12.4657.
Adenovirus 12 (Ad12) (Huie) (highly oncogenic group A) readily induces tumors in newborn rodents. Since Ad12 is isolated from human fecal samples, we investigated whether it plays a role in the etiology of human gastrointestinal cancer. If Ad12 is a causal agent of human cancer, then human tumors should contain Ad12 transforming genes, as indicated by studies of cells transformed in vitro and in vivo by oncogenic viruses. Ad12 DNA and the Ad12 transforming restriction fragment (EcoRI-C fragment, left 16% of the viral genome) were labeled in vitro to 10(7) to 4 X 10(8) cpm/mug by the nick translation reaction of DNA polymerase of Escherichia coli. The fidelity and sensitivity of these probes were established by (i) analysis of DNA from Ad12-transformed cells and from hamsters with tumors induced by Ad12, (ii) reconstruction experiments with added Ad12 DNA and EcoRI restriction fragments, and (iii) comparison of annealing characteristics with Ad12 probes labeled in vivo. With Ad12 [3H]DNA as probe, no viral DNA sequences were detected in 18 normal gastrointestinal tissues and 34 gastrointestinal tumors, including cancers of the colon, rectum, small intestine, and stomach, under conditions that would detect 0.1 copy of the Ad12 genome per tumor cell. Similar analyses of Ad12-transformed hamster cells and Ad12 primary hamster tumors indicated 6-18 copies per cell of over 90% of the viral genome. With the Ad12 EcoRI-C transforming fragment as probe, no hybridization was detected with 32 human gastrointestinal tumors and five normal tissues; this result excludes 1-2% of the Ad12 genome per tumor cell. Our date are strong evidence that Ad12 is not a major cause of human gastrointestinal cancer. The Ad12 transforming EcoRI-C fragment hybridized (50-68% efficiency) with other Ad12 isolates and with Ad18 and 31 (members of oncogenic group A), but not at all with 28 other human Ad serotypes (manuscript in preparation). Thus other group A members probably are also not involved in human gastrointestinal cancer. No viral DNA sequences were detected in 12 normal lungs and 22 lung tumors, suggesting that respiratory cancer does not involve an Ad12 etiology.
腺病毒12型（Ad12）（Huie株）（高致癌性A组）很容易在新生啮齿动物中诱发肿瘤。由于Ad12是从人类粪便样本中分离出来的，我们研究了它是否在人类胃肠道癌症的病因学中起作用。如果Ad12是人类癌症的致病因子，那么人类肿瘤应该含有Ad12转化基因，这已在致癌病毒体外和体内转化细胞的研究中得到证实。通过大肠杆菌DNA聚合酶的缺口平移反应，将Ad12 DNA和Ad12转化限制片段（EcoRI - C片段，病毒基因组左侧16%）在体外标记至10⁷至4×10⁸ cpm/μg。这些探针的保真度和灵敏度通过以下方式确定：（i）分析来自Ad12转化细胞和由Ad12诱发肿瘤的仓鼠的DNA；（ii）用添加的Ad12 DNA和EcoRI限制片段进行重建实验；（iii）将退火特性与体内标记的Ad12探针进行比较。以Ad12 [³H]DNA为探针，在18个正常胃肠道组织和34个胃肠道肿瘤（包括结肠癌、直肠癌、小肠癌和胃癌）中未检测到病毒DNA序列，检测条件为每个肿瘤细胞可检测到0.1个Ad12基因组拷贝。对Ad12转化的仓鼠细胞和Ad12原发性仓鼠肿瘤进行的类似分析表明，每个细胞中超过90%的病毒基因组有6 - 18个拷贝。以Ad12 EcoRI - C转化片段为探针，在32个人类胃肠道肿瘤和5个正常组织中未检测到杂交信号；该结果排除了每个肿瘤细胞中1 - 2%的Ad12基因组。我们的数据有力地证明Ad12不是人类胃肠道癌症的主要病因。Ad12转化的EcoRI - C片段与其他Ad12分离株以及Ad18和31（致癌性A组成员）杂交（效率为50 - 68%），但与其他28种人类腺病毒血清型完全不杂交（正在准备的手稿）。因此，其他A组成员可能也与人类胃肠道癌症无关。在12个正常肺组织和22个肺肿瘤中未检测到病毒DNA序列，这表明呼吸道癌症不涉及Ad12病因。
