SV40 sequences in human osteosarcoma of German origin

Simian virus 40 (SV40) is a dsDNA polyomavirus that induces osteosarcomas a nd mesotheliomas in hamsters and transforms many types of cells in tissue c ulture, including human cells. Osteosarcoma is a bone malignancy with multi ple molecular lesions underlining progression from normal bone to osteosarc oma. Recent investigations have identified SV40 DNA sequences in osteosarco mas, suggesting that SV40 may contribute to tumor development. However, the se studies also demonstrated that geographical differences exist between SV 40 and tumor association. Our study analyzed 46 frozen German tumor specime ns (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 se quences could be detected using the primer set SV.for. 2/SV.rev, while 2 ou t of 42 osteosarcoma specimens and 1 out Of 4 poorly-differentiated tumor s pecimens contained SV40 sequences, using the primer set SV.5/SV.6 From one of these two positive osteosarcomas, multiple tumor biopsies taken at diffe rent 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 geogr aphical differences in the presence of SV40. Finally, the specific detectio n of SV40 sequences with multiple specimens from one of the two patients an d the absence of SV40 sequences in all other samples underscores the specif icity and reproducibility of this investigation and ruled our PCR contamina tion.