The Mutation Profiles of K-RAS/N-RAS Genes in Metastatic Colorectal Cancer Patients

Yıl 2022, , 172 - 178, 03.10.2022

Seda Eren Keskin , Eda Güzdolu , Nilüfer Sertdemir Gülhan Demir Deniz Sünnetçi Akkoyunlu , Devrim Çabuk Naci Çine , Hakan Savlı

https://doi.org/10.30934/kusbed.1052876

Öz

Objective: RAS genes are members of the RAS/Mitogen activated protein kinase pathway which is induced by Epidermal Growth Factor Receptor (EGFR). Mutations in genes in this pathway trigger cancer development. In colorectal cancer, mutations in RAS genes cause resistance to EGRF- targeted therapy. In the treatment of metastatic colorectal cancer, EGFR’s monoclonal antibodies are widely used as chemotherapeutic agents. Kirsten-RAS mutations are found in 30-50% and N-RAS mutations are found in 2-3% of colorectal cancer. In this study, we aimed to analyze Kirsten-RAS /N-RAS mutations in patients with metastatic colorectal cancer.
Methods: One hundred of metastatic colorectal cancer patients resistant to EGFR- targeted therapy were scanned for the Kirsten-RAS mutations status (exon 2,3,4) and N-RAS mutation status (Exon 2,3,4) by Real-Time PCR (Polymerase Chain Reaction) method.
Results: As a result of this study, Kirsten-RAS mutation was found 48% and N-RAS mutation was 1.92%. The most common Kirsten-RAS mutations were in codon 12. The distribution of codon 12 mutations were obtained as G12V (25%), G12D (23%), G12C (14.5%).
Conclusion: In our study, the frequencies of Kirsten-RAS and N-RAS mutations were compitable with similar reports. Our results have supported that testing RAS genes mutations have a vital role in identifying patients who benefit from Epidermal Growth Factor Receptor- targeted therapy.

Anahtar Kelimeler

Metastatic Colorectal Cancer, RAS oncogene, KRAS, NRAS, EGFR-targeted therapy

Metastatik Kolorektal Kanserli Hastalarda KRAS/NRAS Gen Mutasyon Profilleri

Öz

Amaç: RAS genleri, Epidermal Büyüme Faktörü Reseptörü (EGFR) tarafından indüklenen RAS-MAPK Sinyal yolağının bir üyesidir. Bu yolaktaki genlerde meydana gelen mutasyonlar kanser gelişimini tetiklemektedir. Kolorektal kanserde (KRK), RAS genlerinde meydana gelen mutasyonlar EGFR hedefli tedaviye karşı direnç gelişimine neden olur. EGFR monoklonal antikorları, kemoterapötik ajanlar olarak metastatik kolorektal kanser tedavisinde yaygın şekilde kullanılmaktadır. KRAS mutasyonları KRK’nın 30-50%’sinde, NRAS mutasyonları ise 2-3%’ünde bulunur. Bu çalışmada, KRK’lı hastalarda KRAS/NRAS mutasyonlarını analiz etmeyi amaçladık.
Yöntem: EGFR-hedefli tedaviye direnç gösteren 100 metastatik KRK hastası, Real-Time Polimeraz Zincir Reaksiyonu yöntemi ile KRAS mutasyonu (ekzon 2, 3, 4) ve NRAS mutasyonu (ekzon 2, 3, 4) durumu için tarandı.
Bulgular: Bu çalışma sonucunda, KRAS mutasyonu oranı 48% ve NRAS mutasyonu oranı 1,92% olarak bulundu. En yaygın KRAS mutasyonları kodon 12’de saptandı. Kodon 12 mutasyonlarının dağılımı G12V (25%), G12D (23%), G12C (14,5%) olarak elde edildi.
Sonuç: Çalışmamızda saptanan KRAS ve NRAS mutasyon sıklıkları benzer raporlar ile uyumlu bulundu. Sonuçlarımız, RAS mutasyonlarının test edilmesinin EGFR-hedefli tedaviden fayda sağlayacak hastaları belirlemede hayati rolünü desteklemektedir.

Anahtar Kelimeler

KRAS, NRAS, EGRF-hedefli tedavi, Metastatik kolorektal kanser, RAS onkogeni

Kaynakça

• Vogelstein B, Fearon ER, Hamilton SR, Kern SE, Preisinger AC, Leppert M, et al. Genetic alterations during colorectal-tumor development. N Engl J Med. 1988; 319(9): 525-532. doi: 10.1056/NEJM198809013190901.
• Pearson G, Robinson F, Beers Gibson T, Xu BE, Karandikar M, Berman K, et al. Mitogen-activated protein (MAP) kinase pathways: regulation and physiological functions. Endocr Rev. 2001; 22(2): 153-183. doi: 10.1210/edrv.22.2.0428.
• Arrington AK, Heinrich EL, Lee W, Duldulao M, Patel S, Sanchez J, et al. Prognostic and predictive roles of KRAS mutation in colorectal cancer. Int J Mol Sci. 2012; 13(10): 12153-12168. doi:10.3390/ijms131012153.
• Ounissi D, Weslati M, Boughrıba R, Hazgui M, Bouraoui S. Clinicopathological characteristics and mutational profile of KRAS and NRAS inTunisian patients with sporadic colorectal cancer. Turk J of Med Sci. 2021; 51(1): 148-158. doi: 10.3906/sag-2003-42.
• Xie YH, Chen YX, Fang JY. Comprehensive review of targeted therapy for colorectal cancer. Sig Transduct Target Ther. 2020; 22(5): 1-30. doi: 10.1038/s41392-020-0116-z.
• Zhu, G., Pei, L., Xia, H. et al. Role of oncogenic KRAS in the prognosis, diagnosis and treatment of colorectal cancer. Mol Cancer. 2021; 143(20): 1-17. doi: 10.1186/s12943-021-01441-4.
• Colussi D, Brandi G, Bazzoli F, Ricciardiello L. Molecular pathways involved in colorectal cancer: implications for disease behavior and prevention. Int J Mol Sci. 2013; 14(8): 16365-16385. doi: 10.3390/ijms140816365.
• Chang YY, Lin JK, Lin TC, Chen WS, Jeng KJ, Yang SH, et al. Impact of KRAS mutation on outcome of patients with metastatic colorectal cancer. Hepatogastroenterology, 2014; 61(135): 1946–1953.
• Inamura K, Song M, Jung S, Nishihara R, Yamauchi M, Lochhead P, et al. Prediagnosis Plasma Adiponectin in Relation to Colorectal Cancer Risk According to KRAS Mutation Status. J Natl Cancer Inst. 2015; 108(4): djv363. doi: 10.1093/jnci/djv363.
• Ikoma T, Shimokawa M, Kotaka M, Matsumoto T, Nagai H, Boku S et al. Clinical and prognostic features of patients with detailed RAS/BRAF-mutant colorectal cancer in Japan. BMC cancer. 2021; 21(1): 518. doi: 10.1186/s12885-021-08271-z.
• Saharti S. KRAS/NRAS/BRAF Mutation Rate in Saudi Academic Hospital Patients With Colorectal Cancer. Cureus. 2022; 14(4): e24392. doi: 10.7759/cureus.24392.
• Baba O, Bidikian A, Mukherji D, Shamseddin A, Temraz S, Fakhruddin N et al. Tumor profiling of KRAS, BRAF, and NRAS gene mutations in patients with colorectal cancer: A Lebanese major center cohort study. Gene. 2022; 834: 146646. doi: 10.1016/j.gene.2022.146646.
• Hamzehzadeh L, Khadangi F, Ghayoor Karimiani E, Pasdar A, Kerachian MA. Common KRAS and NRAS gene mutations in sporadic colorectal cancer in Northeastern Iranian patients. Curr Probl Cancer. 2018; 42(6): 572-581. doi: 10.1016/j.currproblcancer.2018.05.001.
• Smitha CS, Suresh BMC, Linu JA, Lakshmaiah KC, Govind BK, Lokanatha D, et al. Patterns and the occurrence of KRAS mutations in metastatic colorectal cancers-a study from Indian Regional Cancer Centre. Indian J Surg Oncol. 2017; 8(4): 511-513. doi: 10.1007/s13193-017-0704-8.
• Koçak S. Metastatik kolorektal tümörlerde KRAS ve BRAF mutasyonları ile bevasizumab ve setuksimab tedavisine yanıt arasındaki ilişki. [Uzmanlık Tezi]. İstanbul, Türkiye. İstanbul Üniversitesi Cerrahpaşa Tıp Fakültesi, İç Hastalıkları Anabilim Dalı; 2013.
• American Cancer Society. Colorectal Cancer Facts & Figures 2020-2022, Atlanta: American Cancer Society; 2020. https://www.cancer.org/content/dam/cancer-org/research/cancer-facts-and-statistics/colorectal-cancer-facts-and-figures/colorectal-cancer-facts-and-figures-2020-2022.pdf. Accessed June 28, 2022.
• Wang C, Pan D. Mutation patterns and prognostic analysis of BRAF/KRAS/PIK3CA in colorectal cancer. J Clin Lab Anal. 2022; 36(6); e24444. doi: 10.1002/jcla.24444.
• Alghamdi M, Alabdullatif N, Al-Rashoud A, Alotaibi J, Alhussaini N, Elsirawani S et al. KRAS Mutations in Colorectal Cancer: Relationship With Clinicopathological Characteristics and Impact on Clinical Outcomes in Saudi Arabia. Cureus. 2022; 14(3): e23656. doi: 10.7759/cureus.23656.
• Pylayeva-Gupta Y, Grabocka E, Bar-Sagi D. RAS oncogenes: weaving a tumorigenic web. Nat Rev Cancer. 2011; 11: 761–774. doi:10.1038/nrc3106.
• Dobre M, Dinu DE, Panaitescu E, Bîrlă RD, Iosif CI, Boeriu M, et al. KRAS gene mutations - prognostic factor in colorectal cancer? Rom J Morphol Embryol. 2015; 56: 671-678.
• Neumann J, Zeindl-Eberhart E, Kirchner T, Jung A. Frequency and type of KRAS mutations in routine diagnostic analysis of metastatic colorectal cancer. Pathol Res Pract. 2009; 205(12): 858-862. doi: 10.1016/j.prp.2009.07.010.
• Kawazoe A, Shitara K, Fukuoka S, Kuboki Y, Bando H, Okamoto W, et al. A retrospective observational study of clinicopathological features of KRAS, NRAS, BRAF and PIK3CA mutations in Japanese patients with metastatic colorectal cancer. BMC Cancer. 2015; 15: 258. doi: 10.1186/s12885-015-1276-z.
• Uçar A. 2001-2010 yılları arasında kolorektal karsinom tanılı olguların retrospektif incelenmesi ve KRAS mutasyonunun histopatolojik parametrelerle ilişkisi. [Uzmanlık Tezi]. Antalya, Türkiye. Akdeniz Üniversitesi Tıp Fakültesi Tıbbi Patoloji Anabilim Dalı; 2013.
• Vu Thi MT, Le VT, Huynh QH, Nguyen MD. KRAS gene mutation in patients with primary colorectal cancer. Acta Medica.2019; 50(1), 20-25. doi: 10.32552/ 2019.ActaMedica.337.
• Bruera G, Pepe F, Malapelle U, Pisapia P, Mas AD, Di Giacomo D, et al. KRAS, NRAS and BRAF mutations detected by next generation sequencing, and differential clinical outcome in metastatic colorectal cancer (MCRC) patients treated with first line FIr-B/FOx adding bevacizumab (BEV) to triplet chemotherapy. Oncotarget. 2018; 9(41): 26279-26290. doi: 10.18632/oncotarget.25180.
• Kothari N, Schell MJ, Teer JK, Yeatman T, Shibata D, Kim R. Comparison of KRAS mutation analysis of colorectal cancer samples by standard testing and next-generation sequencing. J Clin Pathol. 2014; 67(9): 764-767. doi:10.1136/jclinpath-2014-202405.
• Tol J, Dijkstra JR, Vink-Börger ME, Nagtegaal ID, Punt CJ, Van Krieken JH, et al. High sensitivity of both sequencing and real-time PCR analysis of KRAS mutations in colorectal cancer tissue. J Cell Mol Med. 2010; 14(8): 2122-2231. doi: 10.1111/j.1582-4934.2009.00788.x.
• Bi F, Wang Q, Dong Q, Wang Y, Zhang L, Zhang J. Circulating tumor DNA in colorectal cancer: opportunities and challenges. Am J Transl Res. 2020; 12(3): 1044-1055.
• 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 systematic review and meta-analysis. Acta Oncol. 2014; 53(7): 852-864. doi: 10.3109/0284186X.2014.895036.
• Meng M, Zhong K, Jiang T, Liu Z, Kwan HY, Su T. The current understanding on the impact of KRAS on colorectal cancer. Biomedicine & pharmacotherapy. 2021; 140: 111717. doi: 10.1016/j.biopha.2021.111717.
• Ostrem JM, Shokat KM. Direct small-molecule inhibitors of KRAS: from structural insights to mechanism-based design. Nat Rev Drug Discov. 2016; 15(11): 771–785. doi: 10.1038/nrd.2016.139.
• Skoulidis F, Li BT, Dy GK, Price TJ, Falchook GS, Wolf J et al. Sotorasib for Lung Cancers with KRAS p.G12C Mutation. NEJM. 2021; 384(25): 2371–2381. doi: 10.1056/NEJMoa2103695.
• Awad MM, Liu S, Rybkin II, Arbour KC, Dilly J, Zhu VW et al. Acquired Resistance to KRASG12C Inhibition in Cancer. NEJM. 2021; 384(25): 2382–2393. doi: 10.1056/NEJMoa2105281.
• Kwan AK, Piazza GA, Keeton AB, Leite CA. The path to the clinic: a comprehensive review on direct KRASG12C inhibitors. J Exp Clin Cancer Res. 2022; 41(1): 27. doi: 10.1186/s13046-021-02225-w.
• Bardelli A, Siena S. Molecular mechanisms of resistance to cetuximab and panitumumab in colorectal cancer. J Clin Oncol. 2010; 28(7): 1254-61. doi: 10.1200/JCO.2009.24.6116.
• Lochhead P, Kuchiba A, Imamura Y, Liao X, Yamauchi M, Nishihara R et al. Microsatellite instability and BRAF mutation testing in colorectal cancer prognostication. J Natl Cancer Inst. 2013; 105(15): 1151-1156. doi: 10.1093/jnci/djt173.
• Sideris M, Adams K, Moorhead J, Diaz-Cano S, Bjarnason I, Papagrigoriadis S. BRAF V600E mutation in colorectal cancer is associated with right-sided tumours and iron deficiency anaemia. Anticancer Res. 2015; 35(4): 2345-2350.
• Nosho K, Kawasaki T, Ohnishi M, Suemoto Y, Kirkner GJ, Zepf D, et al. PIK3CA mutation in colorectal cancer: relationship with genetic and epigenetic alterations. Neoplasia. 2008; 10(6): 534-541. doi: 10.1593/neo.08336.
• Mei ZB, Duan CY, Li CB, Cui L, Ogino S. Prognostic role of tumor PIK3CA mutation in colorectal cancer: a systematic review and meta-analysis. Ann Oncol. 2016; 27(10): 1836-1848. doi: 10.1093/annonc/mdw264.