Advances in Clinical and Experimental Medicine
2019, vol. 28, nr 1, January, p. 67–73
Publication type: original article
Analysis of KRAS, NRAS, BRAF, and PIK3CA mutations could predict metastases in colorectal cancer: A preliminary study
1 Department of Pneumonology, Oncology and Allergology, Medical University of Lublin, Poland
2 Polish Mother's Memorial Hospital Research Institute, Łódź, Poland
3 Genetic and Immunology Institute GENIM Ltd., Lublin, Poland
4 Department of Clinical Patomorphology, Medical University of Lublin, Poland
5 Oncology and Chemotherapy Unit, Medical University of Lublin, Poland
6 Chemotherapy Department, Regional Centre of Oncology, Nicolaus Copernicus Provincial Specialist Hospital, Łódź, Poland
7 Pathology Laboratory Synevo, Łódź, Poland
8 MolGenDia Ltd., Bydgoszcz, Poland
Background. Colorectal cancer (CRC) is usually diagnosed in the metastatic stage, when chemotherapy and molecularly-targeted therapies, instead of surgery, play the most important therapeutic role. Application of anti-epidermal growth factor receptor (EGFR) therapy requires the analysis of RAS mutation status and only RAS wild-type (wt) patients are qualified for the therapy.
Objectives. The objective of this study was to analyze driver mutations in KRAS, NRAS, BRAF, and PIK3CA genes in CRC patients.
Material and Methods. We assessed the KRAS, NRAS, BRAF, and PIK3CA genes in 102 inoperable, locally advanced and advanced CRC patients. Real-time polymerase chain reaction (RT-PCR) and high resolution melt PCR (HRM-PCR) techniques with DNA intercalating dye were applied in the study.
Results. Forty-six patients demonstrated the presence of examined mutations (45.1%). No significant differences in driver mutation occurrence between men and women, as well as between younger (<65 years) and older (≥65 years) patients were found. The mutations were present significantly more frequently in metastatic than in primary tumors (p = 0.039) due to the high incidence of KRAS gene mutations in metastatic tissue. BRAF and PIK3CA mutations were found only in primary tumors. The incidence of PIK3CA mutations was significantly higher (11.77%) in early than in advanced stages of the disease (1.96%; p = 0.05); NRAS mutations were found only in metastatic cancer (7.85%; p = 0.041). Only a single mutation of the PIK3CA and no mutations of NRAS were found in rectal cancer.
Conclusion. Our results have shown low occurrence of driver mutations in Polish CRC patients, involving also mutations in rarely tested genes. The extent of the research panel of additional mutations could contribute to creating a better method of qualifying patients for molecularly targeted therapies and obtaining a better outcome for these therapeutic strategies.
colorectal cancer, PIK3CA, BRAF, molecularly targeted therapy, RAS mutation
- Akkad J, Bochum S, Martens UM. Personalized treatment for colorectal cancer: Novel developments and putative therapeutic strategies. Langenbecks Arch Surg. 2015;400(2):129–143.
- Coppedè F, Lopomo A, Spisni R, Migliore L. Genetic and epigenetic biomarkers for diagnosis, prognosis and treatment of colorectal cancer. World J Gastroenterol. 2014;20(4):943–956.
- Linnekamp JF, Wang X, Medema JP, Vermeulen L. Colorectal cancer heterogeneity and targeted therapy: A case for molecular disease subtypes. Cancer Res. 2015;75(2):245–249.
- Nagtegaal ID, van Krieken JH. Colorectal cancer: Is the new era of colorectal cancer classification finally here? Nat Rev Gastroenterol Hepatol. 2013;10(7):391–393.
- Kim TM, Lee SH, Chung YJ. Clinical applications of next-generation sequencing in colorectal cancers. World J Gastroenterol. 2013;19(40):6784–6793.
- De Sousa E, Melo F, Wang X, et al. Poor-prognosis colon cancer is defined by a molecularly distinct subtype and develops from serrated precursor lesions. Nat Med. 2013;19(5):614–618.
- Vaughn CP, Zobell SD, Furtado LV, Baker CL, Samowittz WS. Frequency of KRAS, NRAS and BRAF mutations in colorectal cancer. Genes Chromosomes Cancer. 2011;50(5):307–312.
- Cathomas G. PIK3CA in colorectal cancer. Front Oncol. 2014;4:35. doi: 10.3389/fonc.2014.00035
- Sartore-Bianchi A, Martini M, Veronese FMS, et al. PIK3CA mutations in colorectal cancer are associated with clinical resistance to EGFR-targeted monoclonal antibodies. Cancer Res. 2009;69(5):1851–1857.
- De Roock W, Claes B, Bernasconi D, et al. Effects of KRAS, BRAF, NRAS and PIK3CA mutations on the efficacy of cetuximab plus chemotherapy in chemotherapy-refractory metastatic colorectal cancer: A retrospective consortium analysis. Lancet Oncol. 2010;11(8):753–762.
- Loupakis F, Ruzzo A, Cremolini C, et al. KRAS codon 61, 146 and BRAF mutations predict resistance to cetuximab plus irinotecan in KRAS codon 12 and 13 wild-type metastatic colorectal cancer. Br J Cancer. 2009;101(4):715–721.
- Van Cutsem E, Lenz HJ, Köhne CH, et al. Fluorouracil, leucovorin, and irinotecan plus cetuximab treatment and RAS mutations in colorectal cancer. J Clin Oncol. 2015;33(7):692–700.
- Bokemeyer C, Kohne CH, Ciardiello F, et al. Treatment outcome according to tumour RAS mutation status in OPUS study patients with metastatic colorectal cancer (mCRC) randomized to FOLFOX4 with/without cetuximab. J Clin Oncol. 2014;32(15 Suppl):abstr 3505.
- Oliner K, Douillard J, Siena S, et al. Analysis of KRAS/NRAS and BRAF mutations in the phase III PRIME study of panitumumab and FOLFOX vs FOLFOX as first line treatment for metastatic colorectal cancer. J Clin Oncol. 2013;31(15 Suppl):abstr 3511.
- Schwartzberg LS, Rivera F, Karthaus M, et al. PEAK: A randomized, multi-centre phase II study of panitumumab plus modified fluorouracil, leucovorin, and oxaliplatin (mFOLFOX6) or bevacizumab plus mFOLFOX6 in patients with previously untreated, unresectable, wild-type KRAS exon 2 metastatic colorectal cancer. J Clin Oncol. 2014;32(21):2240–2247.
- Slattery ML, Curtin K, Wolff RK, et al. A comparison of colon and rectal somatic DNA alterations. Dis Colon Rectum. 2009;52(7):1304–1311.
- Lipsyc M, Yaeger R. Impact of somatic mutations on patterns of metastasis in colorectal cancer. J Gastrointest Oncol. 2015;6(6):645–649.
- Nakayama I, Shinozaki E, Matsushima T, et al. Retrospective study of RAS/PIK3CA/BRAF tumor mutations as predictors of response to first-line chemotherapy with bevacizumab in metastatic colorectal cancer patients. BMC. 2017;17(1):38–46.
- Therkildsen C, Bergmann TK, Henrichsen-Schnack T, Ladelund S, Nilbert M. The predictive value of KRAS, NRAS, BRAF, PIK3CA and PTEN for anti-EGFR treatment in metastatic colorectal cancer: A systemic review and meta-analysis. Acta Oncol. 2014;53(7):852–864.