Green M, Wold W S, Mackey J K, Rigden P
Proc Natl Acad Sci U S A. 1979 Dec;76(12):6606-10. doi: 10.1073/pnas.76.12.6606.
Group C human adenoviruses (Ads) of serotypes 1, 2, 5, and 6 infect most children and commonly cause latent infections of lymphoid tissues. Ads transform cells into a malignant-like phenotype; the oncogenic genetic information is in the left 8% of the viral genome, in the HindIII-G DNA fragment. We have investigated the molecular basis for group C Ad latent infections in human tonsils as well as whether these viruses are linked to human cancer. Tonsil or cancer DNAs and RNAs were assayed for Ad sequences by liquid-phase saturation-hybridization with in vitro-labeled Ad5 HindIII-G fragment. About 25% of the 52 tonsils analyzed contained DNA or RNA sequences specific to HindIII-G, indicating that Ad transforming sequences are expressed as RNA in tonsils. Southern blotting analysis of four tonsil DNAs revealed multiple copies of the complete Ad genome in a free state and provided evidence for an unusual form of the Ad genome, possibly Ad DNA integrated into cellular DNA. In assays of human cancers, no Ad sequences were detected in DNAs from 26 squamous cell carcinomas (Cas), 3 adenocarcinomas, 4 oat cell Cas, 5 stomach Cas, 5 small intestine Cas, 15 colon Cas, 6 rectum Cas, 5 Hodgkin and 6 non-Hodgkin lymphomas, and 2 breast Cas. Reconstruction experiments indicated that the HindIII-G probe could detect 1 copy per cell of 0.2-0.3% of the viral genome. No HindIII-G-specific sequences were detected in RNAs from 21 squamous cell Cas, 3 oat cell Cas, 2 stomach Cas, or 18 colon Cas. In six other experiments using the complete Ad2 genome as probe, no Ad sequences were found in DNAs from 6 lung Cas, 12 normal lung tissues, 33 gastrointestinal Cas, 19 normal gastrointestinal tissues, 6 Hodgkin lymphomas, 3 breast Cas, or 4 kidney Cas, at a sensitivity of about 1 copy per tumor cell of 5-10% of the Ad2 genome. All Ad-induced cancer cells should contain at least 1 copy of 1-6% of the viral genome, the minimal size of the transforming region, and probably should contain multiple copies of more of the genome. Therefore, our data are definitive evidence against group C Ads being the cause of the cancers tested, which represent about 50% of the cancer incidence in the United States. Of additional interest, we did not detect Ad2 sequences in RNAs from 7 human placentas, 12 normal lungs, or 19 normal gastrointestinal tissues (nor in 44 cancer or 23 tonsil RNAs). Thus, we did not confirm a recent report of the presence of Ad2 RNA in RNAs from human placentas; the possibility that a small population of cells in placenta expresses group C "related" sequences is not ruled out.
1、2、5和6型C组人类腺病毒（Ads）感染大多数儿童，并通常导致淋巴组织的潜伏感染。腺病毒可将细胞转化为恶性样表型；致癌基因信息位于病毒基因组左侧8%的HindIII - G DNA片段中。我们研究了人类扁桃体中C组腺病毒潜伏感染的分子基础，以及这些病毒是否与人类癌症有关。通过与体外标记的Ad5 HindIII - G片段进行液相饱和杂交，检测扁桃体或癌症的DNA和RNA中的腺病毒序列。在分析的52个扁桃体中，约25%含有HindIII - G特异的DNA或RNA序列，这表明腺病毒转化序列在扁桃体中以RNA形式表达。对4个扁桃体DNA的Southern印迹分析揭示了游离状态下完整腺病毒基因组的多个拷贝，并为腺病毒基因组的一种异常形式提供了证据，可能是腺病毒DNA整合到细胞DNA中。在人类癌症检测中，在26例鳞状细胞癌（Cas）、3例腺癌、4例燕麦细胞癌、5例胃癌、5例小肠癌、15例结肠癌、6例直肠癌、5例霍奇金淋巴瘤和6例非霍奇金淋巴瘤以及2例乳腺癌的DNA中未检测到腺病毒序列。重建实验表明，HindIII - G探针可检测到每个细胞中病毒基因组0.2 - 0.3%的1个拷贝。在21例鳞状细胞癌、3例燕麦细胞癌、2例胃癌或18例结肠癌的RNA中未检测到HindIII - G特异序列。在另外6个使用完整Ad2基因组作为探针的实验中，在6例肺癌、12例正常肺组织、33例胃肠道癌、19例正常胃肠道组织、6例霍奇金淋巴瘤、3例乳腺癌或4例肾癌的DNA中未发现腺病毒序列，检测灵敏度约为每个肿瘤细胞中Ad2基因组5 - 10%的1个拷贝。所有腺病毒诱导的癌细胞应至少含有病毒基因组1 - 6%的1个拷贝，这是转化区域的最小大小，并且可能应含有更多基因组的多个拷贝。因此，我们的数据明确证明C组腺病毒不是所检测癌症的病因，这些癌症约占美国癌症发病率的50%。另外值得注意的是，我们在7个人类胎盘、12例正常肺组织或19例正常胃肠道组织的RNA中未检测到Ad2序列（在44例癌症或23例扁桃体RNA中也未检测到）。因此，我们没有证实最近关于人类胎盘RNA中存在Ad2 RNA的报道；不排除胎盘中一小部分细胞表达C组“相关”序列的可能性。
