MMTV-like sequences in human breast cancer: a fluorescent PCR/laser microdissection approach.
The hypothesis that a retrovirus homologous to the mouse mammary tumour virus (MMTV) is involved in human breast cancer aetiology has fascinated scientists from many years, but it has never been convincingly demonstrated. Renewed interest in this hypothesis developed when an MMTV env gene-like sequence was found in 38% of human breast cancer tissues. Whereas some subsequent studies confirmed these findings, others did not. The main reasons for this discrepancy, among others, are the different sensitivities and technical details of current molecular approaches to the detection of these sequences. This study is an attempt to find sensitive and reproducible conditions capable of detecting MMTV env-like sequence in human samples. To this end, we first developed a fluorescence nested-PCR (FN-PCR) method that was able to detect very low copies of the viral genome, and then screened a panel of 45 frozen breast cancer samples obtained by laser microdissection. The MMTV env gene-like sequence was found in 15 (33%) of the human breast cancers analysed, whereas the same sequence was detectable neither in normal tissues nor in other types of tumour. Sequence analysis revealed 96% homology with the MMTV genome, but no other significant similarities with the human genome. The combined use of frozen material, microdissected cell populations and FN-PCR provides a novel, sensitive, robust, non-radioactive and fast methodology for the molecular detection of human-MTV. This approach might be successfully used in large molecular studies that aim to investigate the hypothesis of a retroviral aetiology of human breast cancer.
Zammarchi
F
F
Division of Surgical, Molecular and Ultrastructural Pathology, University of Pisa and Pisa University Hospital, Italy.
Pistello
M
M
Piersigilli
A
A
Murr
R
R
Di Cristofano
C
C
Naccarato
A G
AG
Bevilacqua
G
G
D001483:Base Sequence; D001943:Breast Neoplasms / Q000821:virology*; D003001:Cloning, Molecular; D004273:DNA, Neoplasm / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D015752:Genes, env; D006801:Humans; D007834:Lasers; D008324:Mammary Tumor Virus, Mouse / Q000235:genetics*; D042282:Microdissection / Q000379:methods; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods; D012192:Retroviridae Infections / Q000150:complications*; D016415:Sequence Alignment; D017385:Sequence Homology
D004273:DNA, Neoplasm; D004279:DNA, Viral