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Int. J. Mol. Sci. 2016, 17(4), 484; doi:10.3390/ijms17040484

KRAS G12V Mutation Detection by Droplet Digital PCR in Circulating Cell-Free DNA of Colorectal Cancer Patients

1
Health Research Institute-Fundación Jiménez Díaz University Hospital (IIS-FJD), Madrid 28040, Spain
2
Experimental Research Unit, General University Hospital of Albacete, Albacete 02006, Spain
3
Department of Surgery, School of Medicine, Autónoma University of Madrid, Madrid 28029, Spain
4
Department of General Surgery, General Hospital of Villalba, Madrid 28400, Spain
5
Department of Surgery, Fundación Jiménez Díaz University Hospital, Madrid 28040, Spain
*
Author to whom correspondence should be addressed.
Academic Editor: Dario Marchetti
Received: 29 February 2016 / Revised: 21 March 2016 / Accepted: 24 March 2016 / Published: 1 April 2016
(This article belongs to the Special Issue Circulating Tumor Cells)
View Full-Text   |   Download PDF [215 KB, uploaded 1 April 2016]   |  

Abstract

KRAS mutations are responsible for resistance to anti-epidermal growth factor receptor (EGFR) therapy in colorectal cancer patients. These mutations sometimes appear once treatment has started. Detection of KRAS mutations in circulating cell-free DNA in plasma (“liquid biopsy”) by droplet digital PCR (ddPCR) has emerged as a very sensitive and promising alternative to serial biopsies for disease monitoring. In this study, KRAS G12V mutation was analyzed by ddPCR in plasma DNA from 10 colorectal cancer patients and compared to six healthy donors. The percentage of KRAS G12V mutation relative to wild-type sequences in tumor-derived DNA was also determined. KRAS G12V mutation circulating in plasma was detected in 9 of 10 colorectal cancer patients whose tumors were also mutated. Colorectal cancer patients had 35.62 copies of mutated KRAS/mL plasma, whereas in healthy controls only residual copies were found (0.62 copies/mL, p = 0.0066). Interestingly, patients with metastatic disease showed a significantly higher number of mutant copies than M0 patients (126.25 versus 9.37 copies/mL, p = 0.0286). Wild-type KRAS was also significantly elevated in colorectal cancer patients compared to healthy controls (7718.8 versus 481.25 copies/mL, p = 0.0002). In conclusion, KRAS G12V mutation is detectable in plasma of colorectal cancer patients by ddPCR and could be used as a non-invasive biomarker. View Full-Text
Keywords: KRAS; colorectal cancer; plasma; droplet digital PCR; circulating cell-free DNA KRAS; colorectal cancer; plasma; droplet digital PCR; circulating cell-free DNA
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Olmedillas López, S.; García-Olmo, D.C.; García-Arranz, M.; Guadalajara, H.; Pastor, C.; García-Olmo, D. KRAS G12V Mutation Detection by Droplet Digital PCR in Circulating Cell-Free DNA of Colorectal Cancer Patients. Int. J. Mol. Sci. 2016, 17, 484.

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