A novel multiplex RT-PCR system detects human endogenous retrovirus-K in breast cancer

• DOI: 10.1007/s00705-004-0378-8
• Publication Date: 2004-09-20T11:40:21Z
• Authors: H. Davari Ejthadi; J. H. Martin; J. Junying; D. A. Roden; M. Lahiri; P. Warren; P. G. Murray; P. N. Nelson
• Keywords: Biomedical science; Reverse Transcriptase Polymerase Chain Reaction; Human Endogenous Retroviruses; RT-PCR system; Endogenous Retroviruses; 610; Breast Neoplasms; 620; 3. Good health; Life science; 03 medical and health sciences; 0302 clinical medicine; Tumor Cells, Cultured; Medicine; Humans; Female; RNA, Messenger

Human endogenous retrovirus HERV-K like-sequences have been implicated in certain cancers. We developed a novel multiplex RT-PCR system for HERV-K that yielded a 533 bp product together with a smaller sized product (319 bp) of the house keeping gene, histidyl tRNA synthetase (HtRNAS). The latter spanned an intron that also served to validate target cDNA. PCR amplicons of HERV-K and HtRNAS were visualised using a gel documentation system and the pixel intensity used to derive semi-quantitative levels of viral expression. Our data showed that HERV-K10 was significantly elevated in MCF-7 cells treated with estrogen. Interestingly, HERV-K expression was higher in MCF-7 cells selected with adriamycin. RT-PCR combined with Southern blotting also detected HERV-K from breast cancer tissue using laser capture microscopy. This study highlights the presence of HERV-K in the breast cancer cell lines MCF-7 and MCF-7 ADR and confirms HERV-K10 transcripts in the cell line T47D. We believe this study to be a novel approach in determining levels of HERV-K expression and for detecting this virus in cancer cell lines and tissues.