Heinsohn S, Scholz R B, Weber B, Wittenstein B, Werner M, Delling G, Kempf-Bielack B, Setlak P, Bielack S, Kabisch H
Children's University Hospital of Hamburg, Department of Hematology and Oncology, Martinistrasse 52, D-20246 Hamburg, Germany.
Anticancer Res. 2000 Nov-Dec;20(6B):4539-45.
Simian virus 40 (SV40) is a dsDNA polyomavirus that induces osteosarcomas and mesotheliomas in hamsters and transforms many types of cells in tissue culture, including human cells. Osteosarcoma is a bone malignancy with multiple molecular lesions underlining progression from normal bone to osteosarcoma. Recent investigations have identified SV40 DNA sequences in osteosarcomas, suggesting that SV40 may contribute to tumor development. However, these studies also demonstrated that geographical differences exist between SV40 and tumor association. Our study analyzed 46 frozen German tumor specimens (42 osteosarcomas and 4 sarcomas initially suspected to be osteosarcomas) for the presence of SV40 DNA sequences by using PCR. Two different primer sets amplifying a 573 bp region of SV40 Tag gene with the complete intron sequence (SV.for 2/SV.rev) and a 172 bp region with no intron sequence (SV.5/SV.6) were used. DNA sequencing analysis verified the results. No SV40 sequences could be detected using the primer set SV.for 2/SV.rev, while 2 out of 42 osteosarcoma specimens and 1 out of 4 poorly-differentiated tumor specimens contained SV40 sequences, using the primer set SV.5/SV.6. From one of these two positive osteosarcomas, multiple tumor biopsies taken at different times during the dissection, including metastasis, tested positive for SV40. These results indicated that in Germany, only rare osteosarcomas can be linked to SV40. These results support previous findings involving geographical differences in the presence of SV40. Finally, the specific detection of SV40 sequences with multiple specimens from one of the two patients and the absence of SV40 sequences in all other samples underscores the specificity and reproducibility of this investigation and ruled out PCR contamination.
猴空泡病毒40（SV40）是一种双链DNA多瘤病毒，可在仓鼠中诱发骨肉瘤和间皮瘤，并在组织培养中转化多种类型的细胞，包括人类细胞。骨肉瘤是一种骨恶性肿瘤，从正常骨发展到骨肉瘤的过程中有多个分子病变。最近的研究在骨肉瘤中发现了SV40 DNA序列，表明SV40可能与肿瘤发展有关。然而，这些研究也表明SV40与肿瘤的关联存在地理差异。我们的研究通过聚合酶链反应（PCR）分析了46份德国冷冻肿瘤标本（42例骨肉瘤和4例最初怀疑为骨肉瘤的肉瘤）中SV40 DNA序列的存在情况。使用了两组不同的引物，一组扩增包含完整内含子序列的SV40大T抗原基因的573 bp区域（SV.for 2/SV.rev），另一组扩增无内含子序列的172 bp区域（SV.5/SV.6）。DNA测序分析验证了结果。使用引物组SV.for 2/SV.rev未检测到SV40序列，而使用引物组SV.5/SV.6时，42例骨肉瘤标本中有2例以及4例低分化肿瘤标本中有1例含有SV40序列。在这两例阳性骨肉瘤中的一例中，在解剖过程中不同时间采集的多个肿瘤活检样本，包括转移灶，检测出SV40呈阳性。这些结果表明，在德国，只有极少数骨肉瘤可与SV40相关联。这些结果支持了先前关于SV40存在情况的地理差异的研究结果。最后，对两名患者之一的多个样本进行SV40序列的特异性检测，而所有其他样本中均未检测到SV40序列，这突出了本研究的特异性和可重复性，并排除了PCR污染。
