Analysis of human cancers, normal tissues, and verruce plantares for DNA sequences of human papillomavirus types 1 and 2.
Analysis of human cancers, normal tissues, and verruce plantares for DNA sequences of human papillomavirus types 1 and 2.
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对人类癌症、正常组织和跖疣进行 1 型和 2 型人乳头瘤病毒 DNA 序列分析。
DOI：
10.1016/0042-6822(81)90019-2
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发表时间：
1981
期刊：
影响因子：
3.7
通讯作者：
Croissant,O
中科院分区：
文献类型：
--
作者：
Green,M;Orth,G;Wold,WS;Sanders,PR;Mackey,JK;Favre,M;Croissant,O
相对而言，人们对人类乳头瘤病毒（HPV）知之甚少，因为它们不能在组织培养中生长。我们在体外标记了两种HPV病毒的dna， HPV-1是从足底疣中分离出来的，HPV-2是从普通的手疣中分离出来的，并使用这些dna来检查HPV-1和HPV-2之间的同源性，检查乳头瘤病毒病变中HPV基因组的状态，并检测人类癌症中HPV的dna。dna的比活性为5.0× 10.7 ~ 1.1× 108cpm /μg。HPV-1和HPV-2 dna的c0 - t - 12分别为5和7× 10 - 4，与基因组分子量约为5.2× 10 - 6一致。HPV-1和HPV-2 dna的交叉杂交显示只有5-7%的同源性，证实它们是不同的病毒。在检查的10个足底疣中有9个检测到HPV-1 DNA。在分析的9个疣中，没有明确的证据表明8个疣的dna中存在整合的病毒序列。使用这些HPV-1和HPV-2探针，我们对人类癌症dna的HPV序列进行了首次广泛和明确的分子杂交分析。用镍翻译的HPV-1和HPV-2 DNA探针进行饱和杂交，分析人肿瘤DNA的HPV序列。添加HPV-1或HPV-2 DNA的重建实验表明，探针可以检测到每二倍体细胞DNA等效0.1拷贝的病毒基因组。未发现HPV-1序列在人类癌症的dna从156年(14黑色素瘤,3 Ca皮肤,5 Ca咽,1 Ca食道、4 Ca胃、小肠5 Ca, 22 Ca结肠、直肠14 Ca, 25鳞状细胞Ca肺、3肺腺癌,4肺燕麦细胞Ca, 21 Ca肾、膀胱7 Ca, 3 Ca卵巢,3 Ca子宫颈,4 Ca前列腺,10非霍奇金淋巴瘤,2网状细胞肉瘤(脾)),或27日正常人体组织(1皮肤,10扁桃体8结肠癌8肾)。145例人类肿瘤（13例黑色素瘤、4例皮肤癌、2例咽喉癌、3例口腔癌、7例食道癌、4例胃癌、3例小肠癌、29例结肠癌、15例直肠癌、25例肾脏癌、15例膀胱癌、2例卵巢癌、6例宫颈癌、4例前列腺癌、2例精原细胞瘤、11例非霍奇金淋巴瘤）或1例正常人卵巢的dna中未检测到HPV-2序列。这些数据有力地证明，所检测的癌症标本都不是由HPV-1或HPV-2诱导的。然而，需要进一步的工作来充分评估HPV是否可能是人类癌症的病原体，因为本研究中检测的癌症类型仅占美国癌症发病率的50%左右，并且因为我们的探针不会检测到其他已知的HPV类型（HPV-3， HPV-4和HPV-5）的序列。
Comparatively little is known about human papillomaviruses (HPV) because they cannot be grown in tissue culture. We have in vitro labeled DNAs from two HPVs, HPV-1 which was isolated from plantar warts, and HPV-2 which was isolated from common hand warts, and used these DNAs to examine the homology between HPV-1 and HPV-2, to examine the state of the HPV genome in papillomavirus lesions, and to assay human cancer DNAs for HPV. The specific activities of the DNAs were 5.0× 10 7 to 1.1× 10 8 cpm/μg. The C 0 t 1 2 of the HPV-1 and HPV-2 DNAs were 5 and 7× 10− 4, respectively, consistent with a genome molecular weight of about 5.2× 10 6. Cross-hybridization of HPV-1 and HPV-2 DNAs revealed only 5–7% homology, confirming that these are distinct viruses. HPV-1 DNA was detected by Southern blot analysis in 9 of 10 plantar warts examined. No clear evidence was found for integrated viral sequences in DNAs from eight of the nine warts analyzed. Using these HPV-1 and HPV-2 probes, we have performed the first extensive and definitive molecular hybridization analysis of human cancer DNAs for HPV sequences. Human tumor DNAs were analyzed for HPV sequences by saturation hybridization using nick-translated HPV-1 and HPV-2 DNA probes. Reconstruction experiments with added HPV-1 or HPV-2 DNAs indicated that the probes could detect 0.1 copy of the viral genome per diploid equivalent of cellular DNA. No HPV-1 sequences were detected in DNAs from 156 human cancers (14 melanoma, 3 Ca skin, 5 Ca pharynx, 1 Ca esophagus, 4 Ca stomach, 5 Ca small intestine, 22 Ca colon, 14 Ca rectum, 25 squamous cell Ca lung, 3 adenocarcinoma lung, 4 oat cell Ca lung, 21 Ca kidney, 7 Ca bladder, 3 Ca ovary, 3 Ca cervix, 4 Ca prostate, 10 non-Hodgkin lymphoma, 2 reticulum cell sarcoma [spleen]), or 27 normal human tissues (1 skin, 10 tonsil, 8 colon, 8 kidney). No HPV-2 sequences were detected in DNAs from 145 human cancers (13 melanoma, 4 Ca skin, 2 Ca pharynx, 3 Ca mouth, 7 Ca esophagus, 4 Ca stomach, 3 Ca small intestine, 29 Ca colon, 15 Ca rectum, 25 Ca kidney, 15 Ca bladder, 2 Ca ovary, 6 Ca cervix, 4 Ca prostate, 2 Ca seminoma testes, 11 non-Hodgkin lymphoma) or 1 normal human ovary. These data are strong evidence that none of the cancer specimens assayed were induced by HPV-1 or HPV-2. However, additional work is required to fully evaluate whether HPVs are possible agents of human cancers, because the cancer types assayed in this study represent only about 50% of the cancer incidence in the United States, and because our probes would not detect sequences of other recognized HPV types (HPV-3, HPV-4, and HPV-5).