Lack of Aberrant Methylation in an Adjacent Area of Left-Sided Colorectal Cancer
2
조회
0
다운로드
1
인용
0
참고문헌
키워드
초록
Purpose: The molecular nature and the rate-limiting step of epigenetic field defects in the evolution of left-sided colorectal cancer (LCA) remain uncertain. Materials and Methods: The methylation status of 27 candidate field defect markers, six classic CpG island methylator phenotype (CIMP) markers, and LINE-1 were determined in LCA and adjacent normal mucosas (ADJs) from 33 LCA patients and in left normalcolorectal mucosa (LNM) from 33 age- and sex-matched controls. Hotspot mutation analyses in KRAS codons 12 and 13 and BRAF V600E were performed by genomic PCR and pyrosequencing using DNA extracted from endoscopically biopsied tissues. Results: Among the 27 candidate genes tested, we confirmed 15 differentially methylated genes in cancer (15 DMGs; ER, SFRP1, MYOD1, MGMT, CD8a, SPOCK2, ABHD9, BNIP3, IGFBP3, WIF1, MAL, GDNF, ALX4, DOK5, and SLC16A12) in comparison to ADJ samples. We further compared the methylation status of 15 DMGs of ADJs to LNM and found only methylation levels of SLC16A12in ADJs of LCA patients to be significantly higher than that in LNM (17.3% vs. 11.5%, p=0.002). Based on the CIMP, no significantdifferences in methylation levels of the 15 DMGs were found between ADJs in CIMP positive LCA cases and those withoutCIMP. In mutation analyses, no mutation was found in ADJs, while significant KRAS mutations (6/33, 18%) were noted in LCA samples. Conclusion: Epigenetic field defect marked by aberrant methylation is uncommon in normal-appearing ADJs of LCA, indicating the critical rate-limiting change of methylation is likely to occur with morphological alterations in the evolution of LCA.
Abstract
Purpose: The molecular nature and the rate-limiting step of epigenetic field defects in the evolution of left-sided colorectal cancer (LCA) remain uncertain. Materials and Methods: The methylation status of 27 candidate field defect markers, six classic CpG island methylator phenotype (CIMP) markers, and LINE-1 were determined in LCA and adjacent normal mucosas (ADJs) from 33 LCA patients and in left normalcolorectal mucosa (LNM) from 33 age- and sex-matched controls. Hotspot mutation analyses in KRAS codons 12 and 13 and BRAF V600E were performed by genomic PCR and pyrosequencing using DNA extracted from endoscopically biopsied tissues. Results: Among the 27 candidate genes tested, we confirmed 15 differentially methylated genes in cancer (15 DMGs; ER, SFRP1, MYOD1, MGMT, CD8a, SPOCK2, ABHD9, BNIP3, IGFBP3, WIF1, MAL, GDNF, ALX4, DOK5, and SLC16A12) in comparison to ADJ samples. We further compared the methylation status of 15 DMGs of ADJs to LNM and found only methylation levels of SLC16A12in ADJs of LCA patients to be significantly higher than that in LNM (17.3% vs. 11.5%, p=0.002). Based on the CIMP, no significantdifferences in methylation levels of the 15 DMGs were found between ADJs in CIMP positive LCA cases and those withoutCIMP. In mutation analyses, no mutation was found in ADJs, while significant KRAS mutations (6/33, 18%) were noted in LCA samples. Conclusion: Epigenetic field defect marked by aberrant methylation is uncommon in normal-appearing ADJs of LCA, indicating the critical rate-limiting change of methylation is likely to occur with morphological alterations in the evolution of LCA.
참고문헌 (39)
- Slaughter DP, "Field cancerization in oral stratified squamous epithelium; clinical implications of multicentric origin", Cancer, 6, pp.963~968 (1953)
- Braakhuis BJ, "A genetic explanation of Slaughter’s concept of field cancerization:evidence and clinical implications", Cancer Res, 63, pp.1727~1730 (2003)
- Grady WM, "Epigenetic events in the colorectum and in colon cancer", Biochem Soc Trans, 33(Pt 4), pp.684~688 (2005)
- Luo Y, "Field cancerization in the colon: a role for aberrant DNA methylation?", Gastroenterol Rep (Oxf), 2, pp.16~20 (2014)
- Joo Mi Yi, "Epigenetic Alterations in Inflammatory Bowel Disease and Cancer", Intestinal research, 13(2), pp.112~121 (2015), 대한장연구학회
- Baba Y, "Epigenetic field cancerization in gastrointestinal cancers", Cancer Lett, 375, pp.360~366 (2016)
- Baylin SB, "DNA hypermethylation in tumorigenesis:epigenetics joins genetics", Trends Genet, 16, pp.168~174 (2000)
- Feinberg AP, "The epigenetic progenitor origin of human cancer", Nat Rev Genet, 7, pp.21~33 (2006)
- Rashid A, "CpG island methylation in colorectal adenomas", Am J Pathol, 159, pp.1129~1135 (2001)
- Li H, "SLC5A8, a sodium transporter, is a tumor suppressor gene silenced by methylation in human colon aberrant crypt foci and cancers", Proc Natl Acad Sci U S A, 100, pp.8412~8417 (2003)
- Kondo Y, "Epigenetic changes in colorectal cancer", Cancer Metastasis Rev, 23, pp.29~39 (2004)
- Shen L, "MGMT promoter methylation and field defect in sporadic colorectal cancer", J Natl Cancer Inst, 97, pp.1330~1338 (2005)
- Esteller M, "Epigenetic lesions causing genetic lesions in human cancer: promoter hypermethylation of DNA repair genes", Eur J Cancer, 36, pp.2294~2300 (2000)
- Issa JP, "CpG island methylator phenotype in cancer", Nat Rev Cancer, 4, pp.988~993 (2004)
- Issa JP, "Accelerated age-related CpG island methylation in ulcerative colitis", Cancer Res, 61, pp.3573~3577 (2001)
- Clark SJ, "High sensitivity mapping of methylated cytosines", Nucleic Acids Res, 22, pp.2990~2997 (1994)
- Colella S, "Sensitive and quantitative universal pyrosequencing methylation analysis of CpG sites", Biotechniques, 35, pp.146~150 (2003)
- Estécio MR, "LINE-1 hypomethylation in cancer is highly variable and inversely correlated with microsatellite instability", PLoS One, 2, pp.e399~ (2007)
- Ogino S, "Sensitive sequencing method for KRAS mutation detection by pyrosequencing", J Mol Diagn, 7, pp.413~421 (2005)
- Shen L, "Integrated genetic and epigenetic analysis identifies three different subclasses of colon cancer", Proc Natl Acad Sci U S A, 104, pp.18654~18659 (2007)
- Estécio MR, "High-throughput methylation profiling by MCA coupled to CpG island microarray", Genome Res, 17, pp.1529~1536 (2007)
- Chung W, "Identification of novel tumor markers in prostate, colon and breast cancer by unbiased methylation profiling", PLoS One, 3, pp.e2079~ (2008)
- Yamashita S, "Chemical genomic screening for methylation-silenced genes in gastric cancer cell lines using 5-aza-2’-deoxycytidine treatment and oligonucleotide microarray", Cancer Sci, 97, pp.64~71 (2006)
- Hasegawa S, "Genome-wide analysis of gene expression in intestinal-type gastric cancers using a complementary DNA microarray representing 23,040 genes", Cancer Res, 62, pp.7012~7017 (2002)
- Deng D, "Hypermethylation of metallothionein-3 CpG island in gastric carcinoma", Carcinogenesis, 24, pp.25~29 (2003)
- Mikata R, "Analysis of genes upregulated by the demethylating agent 5-aza-2’-deoxycytidine in gastric cancer cell lines", Int J Cancer, 119, pp.1616~1622 (2006)
- Chung SK, "Frequent alteration of XAF1 in human colorectal cancers: implication for tumor cell resistance to apoptotic stresses", Gastroenterology, 132, pp.2459~2477 (2007)
- Ebert MP, "Aristaless-like homeobox-4 gene methylation is a potential marker for colorectal adenocarcinomas", Gastroenterology, 131, pp.1418~1430 (2006)
- Watanabe Y, "PRDM5 identified as a target of epigenetic silencing in colorectal and gastric cancer", Clin Cancer Res, 13, pp.4786~4794 (2007)
- Issa JP, "Methylation of the oestrogen receptor CpG island links ageing and neoplasia in human colon", Nat Genet, 7, pp.536~540 (1994)
- Goel A, "The CpG island methylator phenotype and chromosomal instability are inversely correlated in sporadic colorectal cancer", Gastroenterology, 132, pp.127~138 (2007)
- Clement Richard Boland, "Promoter Methylation in the Genesis of Gastrointestinal Cancer", Yonsei Medical Journal, 50(3), pp.309~321 (2009), 연세대학교의과대학
- Zou H, "Highly methylated genes in colorectal neoplasia: implications for screening", Cancer Epidemiol Biomarkers Prev, 16, pp.2686~2696 (2007)
- Chan AO, "CpG island methylation in aberrant crypt foci of the colorectum", Am J Pathol, 160, pp.1823~1830 (2002)
- Jothy S, "Field effect of human colon carcinoma on normal mucosa: relevance of carcinoembryonic antigen expression", Tumour Biol, 17, pp.58~64 (1996)
- Nguyen C, "Susceptibility of nonpromoter CpG islands to de novo methylation in normal and neoplastic cells", J Natl Cancer Inst, 93, pp.1465~1472 (2001)
- Ushijima T, "DNA methylation as a marker for the past and future", J Gastroenterol, 41, pp.401~407 (2006)
- Ehrlich M, "DNA methylation in cancer: too much, but also too little", Oncogene, 21, pp.5400~5413 (2002)
- Eads CA, "MethyLight: a high-throughput assay to measure DNA methylation", Nucleic Acids Res, 28, pp.E32~ (2000)
