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

• Published in: The Lancet (British edition) Vol. 357; no. 9249; pp. 15 - 20
• Main Authors: Van Trappen, Philippe O, Gyselman, Valerie G, Lowe, David G, Ryan, Andy, Oram, David H, Bosze, Peter, Weekes, Anthony R, Shepherd, John H, Dorudi, Sina, Bustin, Stephen A, Jacobs, Ian J
• Format: Journal Article
• Language: English
• Published: London Elsevier Ltd 06.01.2001; Lancet; Elsevier Limited
• Subjects: Adult; Aged; Base Sequence; Biological and medical sciences; Cervical cancer; Colorectal carcinoma; Female; Follow-Up Studies; Genital system. Mammary gland; Histology; Humans; Investigative techniques, diagnostic techniques (general aspects); Keratins - genetics; Lymph nodes; Lymph Nodes - metabolism; Lymph Nodes - pathology; Lymphatic Metastasis; Lymphatic system; Mapping; Medical sciences; Middle Aged; Molecular biology; Molecular Sequence Data; Neoplasm Recurrence, Local; Neoplasm Staging; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques; Prognosis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; Transcription, Genetic; Tumors; Uterine Cervical Neoplasms - genetics; Uterine Cervical Neoplasms - pathology; Human; Prognosis; RNA-directed DNA polymerase; Lymph node; Enzyme; Transferases; Malignant lymphadenopathy; Early stage; Malignant hemopathy; Tumoral marker; Cytokeratin 19; Uterine cervix; Metastasis; Malignant tumor; Gene expression; Female genital diseases; Polymerase chain reaction; Nucleotidyltransferases; Micrometastasis; Genetics; Diagnosis; Molecular biology; Uterine cervix diseases
• ISSN: 0140-6736; 1474-547X
• DOI: 10.1016/S0140-6736(00)03566-2

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. 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. All primary tumours and histologically involved lymph nodes (six) had more than 10 6 copies of cytokeratin 19 mRNA per μg total RNA. Expression of cytokeratin 19 (up to 1·1×10 5 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 (>10 3 copies) in association with adverse prognostic features. The results suggest that about 50% of early-stage cervical cancers shed tumour cells to the pelvic lymph nodes. The amount of cytokeratin 19 expression was related to clinicopathological features. Further studies are required to document the clinical implications of molecular micrometastases.