Abstract
While it has been nearly 30 years since its discovery, the ras family of genes has not yet lost its impact on basic and clinical oncology. These genes remain central to the field of molecular oncology as tools for investigating carcinogenesis and oncogenic signaling, as powerful biomarkers for the identification of those who have or are at high risk of developing cancer, and as oncogene targets for the design and development of new chemotherapeutic drugs. Mutational activation of the K-RAS proto-oncogene is an early event in the development and progression of the colorectal, pancreatic, and lung cancers that are the major causes of cancer death in the world. The presence of point mutational “hot spots” at sites necessary for the activation of this proto-oncogene has led to the development of a number of highly sensitive PCR-based methods that are feasible for the early detection of K-RAS oncogene mutations in the clinical setting. In light of these facts, mutation at the K-RAS oncogene has the potential to serve as a useful biomarker in the early diagnosis and risk assessment of cancers with oncogenic ras signaling. This chapter describes a highly sensitive method for detecting mutant K-RAS, enriched PCR, and its application to early detection of alterations in this oncogene in preneoplastic and early neoplastic lesions of the colon and rectum.
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References
Hanahan D, Weinberg RA (2000) The hallmarks of cancer. Cell 100:57–70
Vogelstein B, Kinzler KW (2004) Cancer genes and pathways they control. Nat Med 10: 789–799
Ponder BAJ (2001) Cancer genetics. Nature 411:336–341
Malumbres M, Barbacid M (2003) RAS oncogenes; the first 30 years. Nat Rev Cancer 3: 7–13
Cox AD, Der CJ (2002) Ras family signaling. Cancer Biol Ther 1:599–606
Dawnward J (2003) Targeting Ras signalling pathways in cancer therapy. Nat Rev Cancer 3:11–22
Schubbert S, Shannon K, Bollag G (2007) Hyperactive Ras in developmental disorders and cancer. Nat Rev Cancer 7:295–308
Minamoto T, Mai M, Ronai Z (2000) K-ras mutation: early detection in molecular diagnosis and risk assessment of colorectal, pancreas, and lung cancers—a review. Cancer Detect Prev 4:1–12
van Mansfeld ADM, Bos JL (1992) PCR-based approaches for detection of mutated ras genes. PCR Methods Appl 1:211–216
Ronai Z, Yakubovskaya M (1995) PCR in clinical diagnosis. J Clin Lab Anal 9:269–283
Kahn SM, Jiang W, Culbertson TA et al (1991) Rapid and sensitive nonradioactive detection of mutant K-ras genes via ‘enriched’ PCR amplification. Oncogene 6:1079–1083
Jiang W, Kahn S, Guillem J, Lu S, Weinstein IB (1989) Rapid detection of ras oncogenes in human tissues: applications to colon, esophageal, and gastric cancer. Oncogene 4:923–928
Minamoto T, Ronai Z, Yamashita N et al (1994) Detection of Ki-ras mutation in non-neoplastic mucosa of Japanese patients with colorectal cancers. Int J Oncol 4:397–401
Chung DC (2000) The genetic basis of colorectal cancer: insights into critical pathways of tumorigenesis. Gastroenterology 119:864–865
Markowitz SD, Bertagnolli MM (2009) Molecular basis of colorectal cancer. N Engl J Med 361:2449–2460
Minamoto T, Ronai Z (2001) Gene mutation as a target for early detection in cancer diagnosis. Crit Rev Oncol Hematol 40:195–213
Ronai Z, Luo FC, Gradia S, Hart WJ, Butler R (1994) Detection of K-ras mutation in normal and malignant colonic tissues by an enriched PCR method. Int J Oncol 4:391–396
Minamoto T, Yamashita N, Ochiai A et al (1995) Mutant K-ras in apparently normal mucosa of colorectal cancer patients. Its potential as a biomarker of colorectal cancer patients. Cancer 75:1520–1526
Ronai Z, Minamoto T, Butler R et al (1995) Sampling method as a key factor in identifying K-ras oncogene mutations in preneoplastic colorectal lesions. Cancer Detect Prev 19: 512–517
Minamoto T, Esumi H, Ochiai A et al (1997) Combined analysis of microsatellite instability and K-ras mutation increases detection incidence of normal samples from colorectal cancer patients. Clin Cancer Res 3:1413–1417
Tobi M, Luo F-C, Ronai Z (1994) Detection of K-ras mutation in colonic effluent samples from patients without evidence of colorectal carcinoma. J Natl Cancer Inst 86:1007–1010
Zhang B, Ougolkov A, Yamashita K, Takahashi Y, Mai M, Minamoto T (2003) β-catenin and ras oncogenes detect most human colorectal cancers. Clin Cancer Res 3:3073–3079
Alrawi SJ, Schiff M, Carroll RE et al (2006) Aberrant crypt foci. Anticancer Res 26:107–119
Gupta AK, Pretlow TP, Schoen RE (2007) Aberrant crypt foci: what we know and what we need to know. Clin Gastroenterol Hepatol 5:526–533
Yamashita N, Minamoto T, Ochiai A, Onda M, Esumi H (1995) Frequent and characteristic K-ras activation and absence of p53 protein accumulation in aberrant crypt foci of the colon. Gastroenterology 108:434–440
Takayama T, Katsuki S, Takahashi Y et al (1998) Aberrant crypt foci of the colon as precursors of adenoma and cancer. N Engl J Med 339:1277–1284
Gupta AK, Schoerr RE (2009) Aberrant crypt foci: are they intermediate endpoints of colon carcinogenesis in humans? Curr Opin Gastroenterol 25:59–65
Khare S, Chaudhary K, Bissonnette M, Carroll R (2009) Aberrant crypt foci in colon cancer epidemiology. Methods Mol Biol 472:373–386
Otori K, Sugiyama K, Hasebe T, Fukushima S, Esumi H (1995) Emergence of adenomatous aberrant crypt foci (ACF) from hyperplastic ACF with concomitant increase in cell proliferation. Cancer Res 55:4743–4746
Konstantakos AK, Siu I-M, Pretlow TG, Stellato TA, Pretlow TP (1996) Human aberrant crypt foci with carcinoma in situ from a patient with sporadic colon cancer. Gastroenterology 111:772–777
Orlando FA, Tan D, Baltodano JD et al (2008) Aberrant crypt foci as precursors in colorectal cancer progression. J Surg Oncol 98:207–213
Pretlow TP, Brasitus TA, Fulton NC, Cheyer C, Kaplan EL (1993) K-ras mutation in putative preneoplastic lesions in human colon. J Natl Cancer Inst 85:2004–2007
Takayama T, Ohi M, Hayashi T et al (2001) Analysis of K-ras, APC, and β-catenin in aberrant crypt foci in sporadic adenoma, cancer, and familial adenomatous polyposis. Gastroenterology 121:599–611
Suehiro Y, Hinoda Y (2008) Genetic and epigenetic changes in aberrant crypt foci and serrated polyps. Cancer Sci 99:1071–1076
Moore DD, Strauss WM (1995) Preparation of genomic DNA from mammalian tissue. In: Ausubel F, Brent R, Kingston RE et al (eds) Short protocols in molecular biology, 3rd edn. Wiley, Hoboken, NJ, pp 2–8
Wolff R, Gemmill R (1997) DNA from mammalian sources. In: Birren B, Green ED, Klapholz S, Myers RM, Roskams J (eds) Genome analysis: a laboratory manual, vol 1, Cold Spring Harbor Laboratory Press. Plainview, NY, pp 4–16
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Minamoto, T. (2014). Detection and Characterization of Oncogene Mutations in Preneoplastic and Early Neoplastic Lesions. In: Keohavong, P., Grant, S. (eds) Molecular Toxicology Protocols. Methods in Molecular Biology, vol 1105. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-739-6_29
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DOI: https://doi.org/10.1007/978-1-62703-739-6_29
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