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KRAS mutations by digital PCR in circulating tumor cells isolated from the mesenteric vein are associated with residual disease and overall survival in resected colorectal cancer patients

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International Journal of Colorectal Disease Aims and scope Submit manuscript

Abstract

Purpose

In colorectal cancer (CRC), circulating tumor cells (CTCs) are released into the mesenteric veins (MV). We chose to determine whether KRAS mutations detected in CTCs from blood obtained at the time of surgery could be a marker of survival.

Methods

From 52 surgically resected CRC patients who later relapsed, samples of tumor tissue, normal tissue, and blood from the peripheral vein (PV) and MV were obtained from each patient at the time of surgery. KRAS mutations were assessed by Sanger sequencing and digital PCR (DGPCR) in tissue samples and by DGPCR in CTCs. Mutant KRAS copy number was assessed in CTCs. Results were correlated with overall survival (OS).

Results

Sanger sequencing detected KRAS mutations in ten tumor samples (19.2%), while DGPCR detected mutations in 30 (58%). Mutations were detected in CTCs in 21 MV samples (40.4%) and 18 PV samples (34.6%). Patients with G13D mutations in CTCs from the MV had shorter OS than those with G12D mutations (28.1 vs 54.6 months; p = 0.025). Patients with a high mutant KRAS copy number in CTCs had shorter OS than those with a low mutant KRAS copy number (MV: 20.5 vs 43.7 months; p = 0.002; PV: 15.1 vs 38.2 months; p = 0.027).

Conclusion

DGPCR is more efficient than Sanger sequencing for detecting KRAS mutations. KRAS G13D mutations and high mutant KRAS copy number are associated with shorter OS. The analysis of KRAS mutations in CTCs from blood obtained at the time of surgery can identify patients with a higher risk of relapse.

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Funding

This study was partially supported by Servei de Donació del Cos a la Ciencia (Body Donation Service) of the Faculty of Medicine and Health Sciences, University of Barcelona, and by grants from the Ministry of Economy, Industry and Competition, Agencia Estatal de Investigación co-financed with the European Union FEDER funds SAF2017-88606-P (AEI/FEDER, UE).

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Authors and Affiliations

  1. Molecular Oncology and Embryology Laboratory, Human Anatomy Unit, Faculty of Medicine and Health Sciences, University of Barcelona, IDIBAPS, Barcelona, Spain

    Yan Li, Mariano Monzo, Francisco Martinez-Rodenas, Raquel Hernandez, Joan J. Castellano, Jordi Canals, Bing Han, Carmen Muñoz & Alfons Navarro

  2. Department of Medical Oncology and Surgery, Hospital Municipal de Badalona, Badalona, Spain

    Isabel Moreno, Francisco Martinez-Rodenas & Raquel Hernandez

  3. Faculty of Medicine and Health Sciences-Hospital Clinic, University of Barcelona, Casanova 143, ES-08036, Barcelona, Spain

    Alfons Navarro

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  1. Yan Li
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  2. Mariano Monzo
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Correspondence to Alfons Navarro.

Ethics declarations

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional review board of the Hospital Municipal de Badalona and the Ethical Committee of the School of Medicine, University of Barcelona (Institutional Review Board—IRB00003099) and with the 1964 Helsinki Declaration and its later amendments. Informed consent was obtained from all individual participants included in the study. Y.L and J.C are APIF fellows of the Universitat de Barcelona. The other authors declare no conflict of interest.

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Li, Y., Monzo, M., Moreno, I. et al. KRAS mutations by digital PCR in circulating tumor cells isolated from the mesenteric vein are associated with residual disease and overall survival in resected colorectal cancer patients. Int J Colorectal Dis 35, 805–813 (2020). https://doi.org/10.1007/s00384-020-03538-6

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