Research Papers: Pathology:
Sensitive molecular detection of small nodal metastasis in uterine cervical cancer using HPV16-E6/CK19/MUC1 cancer biomarkers
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Vanessa Samouëlian1,2,3, Nawel Mechtouf1, Eric Leblanc4, Guillaume B. Cardin1, Valérie Lhotellier5, Denis Querleu6, Françoise Révillion5,* and Francis Rodier1,7,*
1CRCHUM et Institut du cancer de Montréal, Montreal, QC, Canada
2Université de Montréal, Département d’Obstétrique Gynécologie, Montreal, QC, Canada
3CHUM, Service de Gynécologie oncologique, Montreal, QC, Canada
4Department of Surgery - Centre Oscar Lambret, Lille Cedex, France
5Laboratory of Human Molecular Oncology - Centre Oscar Lambret, Lille Cedex, France
6Institut Bergonie, Bordeaux, France
7Université de Montréal, Département de Radiologie, Radio-Oncologie et Médicine Nucléaire, Montreal, QC, Canada
*These authors contributed equally to this work
Françoise Révillion, email: [email protected]
Vanessa Samouëlian, email: [email protected]
Keywords: diagnostic of lymph node metastasis; HPV viral oncogenes; intraoperative pcr; pretherapeutic evaluation; RT-PCR; Pathology
Received: May 01, 2016 Accepted: March 15, 2018 Published: April 24, 2018
Metastatic nodal involvement is a critical prognostic factor in uterine cervical cancer (UCC). To improve current methods of detecting UCC metastases in lymph nodes (LNs), we used quantitative PCR (qPCR) to assess mRNA expression of potential metastatic biomarkers. We found that expression of HPV16-E6, cytokeratin19 (CK19), and mucin1 (MUC1) is consistently upregulated in tumors and metastatic tissues, supporting a role for these genes in UCC progression. These putative biomarkers were able to predict the presence of histologically positive metastatic LNs with respective sensitivities and specificities of 82% and 99% (CK19), 76% and 95% (HPV16-E6), and 76% and 78% (MUC1). While the biomarkers failed to detect 1.7% to 2.2% of the histologically positive LNs when used individually, combining CK19 and HPV16-E6 enhanced sensitivity and specificity to 100% and 94%, respectively. To explore the sensitivity of qPCR-based detection of varying proportions of invading HPV16-positive UCC cells, we designed a LN metastasis model that achieved a fresh cell detection limit of 0.008% (1:12500 HPV16-positive to HPV16-negative cells), and a paraffin-embedded, formalin-fixed (PEFF) detection limit of 0.02% (1:5000 HPV16-positive to HPV16-negative cells), both of which are within the theoretical detection limit for micrometastasis. Thus, HPV E6/E7 oncogenes may be useful targets for the ultrasensitive detection of nodal involvements like micrometastases in fresh or archived tissue samples. Moreover, our results suggest that the biomarker combination of CK19/HPV-E6 could support a real-time intraoperative strategy for the detection of small, but potentially lethal, metastatic nodal involvements in fresh UCC tissues.
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