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Detection and characterization of viral genomes and search for tumoral antigens in two hamster cell lines derived from tumors induced by bovine papillomavirus type 1
Author(s) -
Breitburd FrançOise,
Favre Michel,
Zoorob Rima,
Fortin Dominique,
Orth Gérard
Publication year - 1981
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910270517
Subject(s) - biology , microbiology and biotechnology , dna , hamster , restriction enzyme , cell culture , bovine papillomavirus , genome , virology , cell , southern blot , biochemistry , genetics , gene
Two cell lines, designated HT2 and HT3, were isolated from two transplanted hamster sarcomas induced by bovine papillomavirus type I (BPV‐I) and studied after 50 to 60 subcultures. Cells are tumorigenic for hamsters and showed no detectable viral capsid protein synthesis by immunofluorescence technique. Cells contained multiple copies of the entire BPV‐I genome, 100 to 500 and 10 to 100 per diploid amount of HT2 and HT3 cell DNA, respectively, as determined by blot hybridization experiments. In unrestricted cell DNA, viral genomes migrated as free single viral DNA molecules and slow migrating species. HT2 cell DNA restricted by single cut enzymes for BPV‐I DNA (Eco RI and Bam HI endonucleases) yielded only one viral genome unit length DNA molecule, indicating that the slow migrating species could correspond to free oligomeric or catenated molecules. Patterns of HT3 cell DNA restricted by Eco RI or Bam HI enzymes showed the presence of additional bands, suggesting that the slow migrating species could correspond to viral DNA sequences integrated in cell DNA. Up to five polypeptides (190 K, 125 K, 59 K, 33 K, 30 K) were reproducibly precipitated from [ 35 S]‐methionine‐ or 32 P ‐ labelled HT2 or HT3 cell extracts by sera from hamsters bearing HT2 or HT3 cell‐induced tumors and by sera from pikas bearing BPV‐I induced fibrosarcomas. Polypeptides with similar molecular weights were detected by HT2 sera in a SV40 tumor hamster cell line (TSV5 CI.2) and in a rat cell line transformed in vitro by SV40 DNA form I (VII FI CI. I). The 190 K polypeptide was also detected in a normal hamster embryo cell strain (EH). Conversely, a pool of SV40 tumor hamster sera precipitated the 59 K and the 33 K polypeptides from HT2 cells. The polypeptides thus identified are most probably transformation‐related cell proteins. Whether or not their synthesis is related to the expression of some BPV‐I transforming genes remains to be established.