Molecular quantification and mapping of lymph-node micrometastases in cervical cancer

Philippe O. Van Trappen, Valerie G. Gyselman, David G. Lowe, Andy Ryan, David H. Oram, Peter Bosze, Anthony R. Weekes, John H. Shepherd, Sina Dorudi, Stephen A. Bustin, Ian J. Jacobs

    Research output: Contribution to journalArticlepeer-review

    Abstract

    Background: A proportion of patients with cancer and lymph nodes negative on histology will develop recurrence. Reverse-transcriptase PCR (RT-PCR) is a highly sensitive method for detection of lymph-node micrometastases, but accurate quantitative assessment has been difficult. Methods: We studied primary tumours and 156 lymph nodes from 32 patients with cervical cancer (stage IA2, IB1, and IB2) and 32 lymph nodes from nine patients with benign disease. A fully quantitative, real-time RT-PCR assay was used to document absolute copy numbers of the epithelial marker cytokeratin 19. Primers and probe were designed not to amplify either of the two cytokeratin 19 pseudogenes. Findings: All primary tumours and histologically involved lymph nodes (six) had more than 106 copies of cytokeratin 19 mRNA per μg total RNA. Expression of cytokeratin 19 (up to 1.1×105 copies per μg RNA) was detected in 66 (44%) of 150 histologically uninvolved lymph nodes, and in nodes from 16 of 32 patients with cervical cancer. 15 of these 16 patients with evidence of micrometastases had the highest cytokeratin 19 transcription level in a first lymph-node drainage station (three obturator, six internal, and six external iliac node). Transcription of cytokeratin 19 was found at a low level in just one of 32 lymph nodes obtained from nine patients with benign disease. Median copy number of cytokeratin 19 transcription was significantly higher (
    Original languageEnglish
    Pages (from-to)15-20
    Number of pages5
    JournalThe Lancet
    Volume357
    Issue number9249
    DOIs
    Publication statusPublished - 6 Jan 2001

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