食管癌中CHD5基因表观遗传学改变
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摘要
背景
食管癌是世界上最常见8大的恶性肿瘤之一,近20年来其发病率在全球范围内呈上升趋势,每年约380,000患者死于该病。但是,食管癌的发病机制至今仍不清楚。最近研究表明,基因启动子区的异常甲基化导致基因表达沉默,可能参与人类肿瘤的发病过程。
染色质解旋酶DNA结合蛋白5(chromodomain helicase DNA-binding protein5)基因是chromodomain超家族的一员,是2003年新发现的抑癌基因。CHD5基因启动子区异常甲基化导致该基因沉默在不同的人类肿瘤中已有报道,但其在食管癌发生中的表观遗传学变化及调控机制尚不清楚。
目的：
研究食管鳞癌组织和食管癌细胞系KYSE70、KYSE140、SKGT4和YES2中CHD5基因表观遗传学改变,探讨CHD5基因甲基化作为食管癌诊断标志物的可行性。
方法：
用甲基化特异性聚合酶链反应(methylation specific PCR,MSP)检测72例食管鳞癌组织及成对癌旁组织,9例正常食管粘膜组织及4株食管癌细胞系中CHD5基因的甲基化状态,并用逆转录聚合酶链式反应(reverse transcriptionPCR,RT-PCR)检测CHD5基因在上述食管癌细胞系的表达。本实验采用SPSS17.0统计软件包进行统计分析,以a=0.05为显著检验水准。
结果：
69%(50/72)食管鳞癌组织发生CHD5基因启动子区甲基化,32%(23/72)癌旁组织发生甲基化,差异具有统计学意义(x2=20.254,P<0.05)。9例食管正常组织未发生甲基化,2株食管癌细胞系KYSE70、SKGT4中由于基因启动子区甲基化导致CHD5基因表达缺失,经5-aza-dc处理96h后CHD5重新表达。
结论：
CHD5基因在食管鳞癌中频繁发生甲基化；CHD5基因mRNA在食管癌细胞系中受甲基化调控,表观遗传学改变是其基因表达及转录的重要调节机制。
Background:
Esophageal cancer ranks as the8th most common cancer worldwide. The incidence of esophageal cancer increased rapidly in the past twenty years and about380,000cases died each year. However, the mechanisms contributing to carcinogenesis of esophageal cancer are still poorly understood. Recent studies showed that aberrant promoter DNA methylation contributes to gene silencing, may participate in the carcinogenesis of human cancer.
Chromodomain helicase DNA-binding protein5(CHD5) is a member of chromodomain. It was been discovered as a novel tumor suppressor gene in2003. Promoter region methylation is related to loss of CHD5expression has been found in many human cancer. However, CHD5epigenetic changes in esophageal cancer remains unclear.
Objective:
To explore epigenetic changes of CHD5during esophageal squamous cell carcinoma and the possibility of CHD5promoter region methylation as human esophageal cancer marker.
Method:
Methylation-specific polymerase chain reaction (MSP) was used to detect the methylation status of CHD5in72cases esophageal squamous cell carcinoma and matched adjacent tissue, nine cases normal mucosa, and four esophageal cancer cell linesKYSE70, KYSE140, SKGT4and YES2. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the expression of CHD5in esophageal cancer cell lines mentioned above. In this topic we use SPSS17.0statistical software package for statistical analysis, and the significant test standard was a=0.05.
Results:
69%(50/72) cases of esophageal squamous cell carcinoma were methylated, and32%(23/72)cases were methylated in matched adjacent tissue(x2=20.254, P<0.05). Nine cases of normal esophageal mucosa were unmethylated. Loss of CHD5expression was found in2esophageal cancer cell lines KYSE70and SKGT4with promoter region methylation, and after treatment with5-aza-dc for96h, the CHD5was re-expression.
Conclusion:
CHD5is frequently methylated in esophageal squamous cell carcinoma; The mRNA of CHD5is regulated by DNA methylatuion in esophageal cancer, epigenetic change may be an important regulation mechanism of its expression and transcription.
食管癌是世界上最常见8大的恶性肿瘤之一,近20年来其发病率在全球范围内呈上升趋势,每年约380,000患者死于该病。但是,食管癌的发病机制至今仍不清楚。最近研究表明,基因启动子区的异常甲基化导致基因表达沉默,可能参与人类肿瘤的发病过程。
染色质解旋酶DNA结合蛋白5(chromodomain helicase DNA-binding protein5)基因是chromodomain超家族的一员,是2003年新发现的抑癌基因。CHD5基因启动子区异常甲基化导致该基因沉默在不同的人类肿瘤中已有报道,但其在食管癌发生中的表观遗传学变化及调控机制尚不清楚。
目的：
研究食管鳞癌组织和食管癌细胞系KYSE70、KYSE140、SKGT4和YES2中CHD5基因表观遗传学改变,探讨CHD5基因甲基化作为食管癌诊断标志物的可行性。
方法：
用甲基化特异性聚合酶链反应(methylation specific PCR,MSP)检测72例食管鳞癌组织及成对癌旁组织,9例正常食管粘膜组织及4株食管癌细胞系中CHD5基因的甲基化状态,并用逆转录聚合酶链式反应(reverse transcriptionPCR,RT-PCR)检测CHD5基因在上述食管癌细胞系的表达。本实验采用SPSS17.0统计软件包进行统计分析,以a=0.05为显著检验水准。
结果：
69%(50/72)食管鳞癌组织发生CHD5基因启动子区甲基化,32%(23/72)癌旁组织发生甲基化,差异具有统计学意义(x2=20.254,P<0.05)。9例食管正常组织未发生甲基化,2株食管癌细胞系KYSE70、SKGT4中由于基因启动子区甲基化导致CHD5基因表达缺失,经5-aza-dc处理96h后CHD5重新表达。
结论：
CHD5基因在食管鳞癌中频繁发生甲基化；CHD5基因mRNA在食管癌细胞系中受甲基化调控,表观遗传学改变是其基因表达及转录的重要调节机制。
Background:
Esophageal cancer ranks as the8th most common cancer worldwide. The incidence of esophageal cancer increased rapidly in the past twenty years and about380,000cases died each year. However, the mechanisms contributing to carcinogenesis of esophageal cancer are still poorly understood. Recent studies showed that aberrant promoter DNA methylation contributes to gene silencing, may participate in the carcinogenesis of human cancer.
Chromodomain helicase DNA-binding protein5(CHD5) is a member of chromodomain. It was been discovered as a novel tumor suppressor gene in2003. Promoter region methylation is related to loss of CHD5expression has been found in many human cancer. However, CHD5epigenetic changes in esophageal cancer remains unclear.
Objective:
To explore epigenetic changes of CHD5during esophageal squamous cell carcinoma and the possibility of CHD5promoter region methylation as human esophageal cancer marker.
Method:
Methylation-specific polymerase chain reaction (MSP) was used to detect the methylation status of CHD5in72cases esophageal squamous cell carcinoma and matched adjacent tissue, nine cases normal mucosa, and four esophageal cancer cell linesKYSE70, KYSE140, SKGT4and YES2. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the expression of CHD5in esophageal cancer cell lines mentioned above. In this topic we use SPSS17.0statistical software package for statistical analysis, and the significant test standard was a=0.05.
Results:
69%(50/72) cases of esophageal squamous cell carcinoma were methylated, and32%(23/72)cases were methylated in matched adjacent tissue(x2=20.254, P<0.05). Nine cases of normal esophageal mucosa were unmethylated. Loss of CHD5expression was found in2esophageal cancer cell lines KYSE70and SKGT4with promoter region methylation, and after treatment with5-aza-dc for96h, the CHD5was re-expression.
Conclusion:
CHD5is frequently methylated in esophageal squamous cell carcinoma; The mRNA of CHD5is regulated by DNA methylatuion in esophageal cancer, epigenetic change may be an important regulation mechanism of its expression and transcription.
引文
[1]Kamangar F, Dores G M, Anderson W F. Patterns of cancer incidence, mortality, and prevalence across five continents:defining priorities to reduce cancer disparities in different geographic regions of the world[J]. J Clin Oncol,2006,24(14):2137-2150.
[2]Jemal A, Siegel R, Ward E, et al. Cancer statistics,2009[J]. CA Cancer J Clin, 2009,59(4):225-249.
[3]Guo M, Ren J, Brock M V, et al. Promoter methylation of HIN-1 in the progression to esophageal squamous cancer[J]. Epigenetics,2008,3(6):336-341.
[4]Holliday R. The inheritance of epigenetic defects[J]. Science,1987,238(4824):163-170.
[5]Matias K P. Study of epigenetic changes leads to treatment advances in adult leukemias.Oncol.2003;48:1-4.
[6]张晓梅Dact2在肝癌中的表观遗传学变化及功能研究[D].军医进修学院解放军总医院解放军总医院解放军军医进修学院消化系病,2010.
[7]尹东涛.SOX17在肺癌中的表观遗传改变及功能研究[D].解放军总医院军医进修学院解放军总医院军医进修学院胸外科,2011.
[8]黄庆,郭颖,府伟灵.人类表观基因组计划[J].生命的化学,2004,24(2)：101-103.
[9]Hermann A, Gowher H, Jeltsch A. Biochemistry and biology of mammalian DNA methyltransferases[J]. Cell Mol Life Sci,2004,61(19-20):2571-2587.
[10]王新刚,王笑峰,李福年,等.表观遗传学药物影响乳腺癌细胞系抑癌基因BRCA1及CHD5表达的研究[J].山东大学学报(医学版),2011,49(7)：29-34.
[11]White P S, Thompson P M, Gotoh T, et al. Definition and characterization of a region of 1p36.3 consistently deleted in neuroblastoma [J]. Oncogene,2005,24(16):2684-2694.
[12]Mulero-Navarro S, Esteller M. Chromatin remodeling factor CHD5 is silenced by promoter CpG island hypermethylation in human cancer[J]. Epigenetics,2008,3(4):210-215.
[13]韩琳琳,侯琳,宋金莲.甲硫氨酸合成酶多态性及CHD5基因甲基化与乳腺癌发病的关系[J].中国癌症杂志,2010,20(4)：251-255.
[14]Garber K. Momentum building for human epigenome project[J]. J Natl Cancer Inst, 2006,98(2):84-86.
[15]Holliday R. Epigenetics:an overview[J]. Dev Genet,1994,15(6):453-457.
[16]Smits K M, Cleven A H, Weijenberg M P, et al. Phannacoepigenomics in colorectal cancer:a step forward in predicting prognosis and treatment response[J]. Pharmacogenomics, 2008,9(12):1903-1916.
[17]Jones PA, Laird PW. Cancer epigenetics comes of age[J]. Nat Genet,1999,21(2):163-167.
[18]Paz M F, Fraga M F, Avila S, et al. A systematic profile of DNA methylation in human cancer cell lines[J]. Cancer Res,2003,63(5):1114-1121.
[19]Gama-Sosa M A, Slagel V A, Trewyn R W, et al. The 5-methylcytosine content of DNA from human tumors[J]. Nucleic Acids Res,1983,11(19):6883-6894.
[20]Gonzalez-Zulueta M, Bender C M, Yang A S, et al. Methylation of the 5'CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing[J]. Cancer Res,1995,55(20):4531-4535.
[21]Herman J G, Merlo A, Mao L, et al. Inactivation of the CDKN2/p16/MTS1 gene is frequently associated with aberrant DNA methylation in all common human cancers[J]. Cancer Res, 1995,55(20):4525-4530.
[22]Sudarsanam P, Winston F. The Swi/Snf family nucleosome-remodeling complexes and transcriptional control[J]. Trends Genet,2000,16(8):345-351.
[23]Hall J A, Georgel P T. CHD proteins:a diverse family with strong ties[J]. Biochem Cell Biol, 2007,85(4):463-476.
[24]Bagchi A, Mills A A. The quest for the 1p36 tumor suppressor[J]. Cancer Res, 2008,68(8):2551-2556.
[25]Isidor B, Le Cunff M, Boceno M, et al. Complex constitutional subtelomeric 1p36.3 deletion/duplication in a mentally retarded child with neonatal neuroblastoma[J]. Eur J Med Genet,2008,51(6):679-684.
[26]Okawa E R, Gotoh T, Manne J, et al. Expression and sequence analysis of candidates for the 1p36.31 tumor suppressor gene deleted in neuroblastomas[J]. Oncogene, 2008,27(6):803-810.
[27]Maser R S, Choudhury B, Campbell P J, et al. Chromosomally unstable mouse tumours have genomic alterations similar to diverse human cancers[J]. Nature,2007,447(7147):966-971.
[28]Bagchi A, Papazoglu C, Wu Y, et al. CHD5 is a tumor suppressor at human lp36[J]. Cell, 2007,128(3):459-475.
[29]Zhang C, Li H, Wang Y, et al. Epigenetic inactivation of the tumor suppressor gene RIZ1 in hepatocellular carcinoma involves both DNA methylation and histone modifications [J]. J Hepatol,2010,53(5):889-895.
[30]Smith L T, Lin M, Brena R M, et al. Epigenetic regulation of the tumor suppressor gene TCF21 on 6q23-q24 in lung and head and neck cancer[J]. Proc Natl Acad Sci USA, 2006,103(4):982-987.
[31]Lin J, Beerm D G. Molecular biology of upper gastrointestinal malignancies[J]. Semin Oncol, 2004,31(4):476-486.
[32]Guo M, House M G, Akiyama Y, et al. Hypermethylation of the GATA gene family in esophageal cancer[J]. Int J Cancer,2006,119(9):2078-2083.
[33]Tzao C, Hsu H S, Sun G H, et al. Promoter methylation of the hMLH 1 gene and protein expression of human mutL homolog 1 and human mutS homolog 2 in resected esophageal squamous cell carcinoma[J]. J Thorac Cardiovasc Surg,2005,130(5):1371.
[34]Okawa E R, Gotoh T, Manne J, et al. Expression and sequence analysis of candidates for the 1p36.31 tumor suppressor gene deleted in neuroblastomas[J]. Oncogene, 2008,27(6):803-810.
[1]Jemal A, Siegel R, Ward E, et al. Cancer statistics,2009[J]. CA Cancer J Clin, 2009,59(4):225-249.
[2]Kanavos P. The rising burden of cancer in the developing world[J]. Ann Oncol,2006,17 SuPp18:i15-i23.
[3]Kamangar F, Dores G M, Anderson W F. Patterns of cancer incidence, mortality, and prevalenceacross five continents:defining priorities to reduce cancer disparities in different geographic regions of the world [J]. J Clin Oncol,2006,24 (14):2137-2150.
[4]Pohl H, Welch H G. The role of overdiagnosis and reclassification in the marked increase of esophageal adenocarcinoma incidence[J]. J Natl Cancer Inst,2005,97(2):142-146.
[5]Holliday R. Epigenetics:an overview[J]. Dev Genet,1994,15(6):453-457.
[6]Gama-Sosa M A, Midgett R M, Slagel V A, et al. Tissue-specific differences in DNA methylation in various mammals[J]. Biochim Biophys Acta,1983,740(2):212-219.
[7]Gonzalez-Zulueta M, Bender C M, Yang A S, et al. Methylation of the 5'CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing[J]. Cancer Res,1995,55(20):4531-4535.
[8]Herman J G, Latif F, Weng Y, et al. Silencing of the VHL tumor-suppressor gene by DNA methylation in renal carcinoma[J]. Proc Natl Acad Sci USA,1994,91 (21):9700-9704.
[9]Reik W, Lewis A. Co-evolution of X-chromosome inactivation and imprinting in mammals[J]. Nat Rev Genet,2005,6(5):403-410.
[10]Ramsahoye B H, Burnett A K, Taylor C. Restriction endonuclease isoschizomers Ital, BsoFI and Fsp4HI are characterised by differences in their sensitivities to CpG methylation[J]. Nucleic Acids Res,1997,25(16):3196-3198.
[11]Saxonov S, Berg P, Brutlag D L. A genome-wide analysis of CpG dinucleotides in the human genome distinguishes two distinct classes of promoters[J]. Proc Natl Acad Sci U SA,2006,103(5):1412-1417.
[12]Kulis M, Esteller M. DNA methylation and cancer[J]. Adv Genet,2010,70:27-56.
[13]Kato Y, Kaneda M, Hata K, et al. Role of the Dnmt3 family in de novo methylation of imprinted and repetitive sequences during male germ cell development in the mouse[J]. Hum Mol Genet,2007,16(19):2272-2280.
[14]Lin H, Yamada Y, Nguyen S, et al. Suppression of intestinal neoplasia by deletion of Dnmt3b[J]. Mol Cell Biol,2006,26(8):2976-2983.
[15]Deplus R, Brenner C, Burgers W A, et al. Dnmt3L is a transcriptional repressor that recruits histone deacetylase[J]. Nucleic Acids Res,2002,30(17):3831-3838.
[16]Baba Y, Huttenhower C, Nosho K, et al. Epigenomic diversity of colorectal cancer indicated by LINE-1 methylation in a database of 869 tumors[J]. Mol Cancer, 2010,9:125.
[17]白志勇,徐国宾,武淑兰.同位素示踪微量法测定白血病细胞DNA-甲基转移酶活性[J].北京医科大学学报,2000,32(1)：76-78.
[18]郑全辉,张宗玉,童坦君.DNA甲基化在细胞衰老中的作用[J].生命的化学,2003,23(4)：263-265.
[19]Bird A P. The relationship of DNA methylation to cancer[J]. Cancer Surv, 1996,28:87-101.
[20]Galm O, Herman J G, Baylin S B. The fundamental role of epigenetics in hematopoietic malignancies[J]. Blood Rev,2006,20(1):1-13.
[21]Seifert H H, Schmiemann V, Mueller M, et al. In situ detection of global DNA hypomethylation in exfoliative urine cytology of patients with suspected bladder cancer[J]. Exp Mol Pathol,2007,82(3):292-297.
[22]郭晓青,李兆中.DNA甲基化与恶性肿瘤[J].国际消化病杂志,2007,27(1)：57-60.
[23]Wu K, Weng Z, Tao Q, et al. Stage-specific expression of breast cancer-specific gene gamma-synuclein[J]. Cancer Epidemiol Biomarkers Prev,2003,12(9):920-925.
[24]Milicic A, Harrison L A, Goodlad R A, et al. Ectopic expression of P-cadherin correlates with promoter hypomethylation early in colorectal carcinogenesis and enhanced intestinal crypt fission in vivo[J]. Cancer Res,2008,68(19):7760-7768.
[25]Liu H, Liu W, Wu Y, et al. Loss of epigenetic control of synuclein-gamma gene as a molecular indicator of metastasis in a wide range of human cancers[J]. Cancer Res, 2005,65(17):7635-7643.
[26]Roman-Gomez J, Jimenez-Velasco A, Agirre X, et al. Promoter hypomethylation of the LINE-1 retrotransposable elements activates sense/antisense transcription and marks the progression of chronic myeloid leukemia[J]. Oncogene,2005,24(48):7213-7223.
[27]Buchi F, Spinelli E, Masala E, et al. Proteomic analysis identifies differentially expressed proteins in AML1/ETO acute myeloid leukemia cells treated with DNMT inhibitors azacitidine and decitabine[J]. Leuk Res,2012,36(5):607-618.
[28]Saito Y, Kanai Y, Sakamoto M, et al. Overexpression of a splice variant of DNA methyltransferase 3b, DNMT3b4, associated with DNA hypomethylation on pericentromeric satellite regions during human hepatocarcinogenesis[J]. Proc Natl Acad Sci USA,2002,99(15):10060-10065.
[29]Gonzalez-Gomez P, Bello M J, Alonso M E, et al. CpG island methylation status and mutation analysis of the RB1 gene essential promoter region and protein-binding pocket domain in nervous system tumours[J]. Br J Cancer,2003,88(1):109-114.
[30]侯志平,何培元,周伟娜.甲基化与肿瘤的相关性研究[J].承德医学院学报,2010,27(2)：194-196.
[31]Weller M, Stupp R, Reifenberger G, et al. MGMT promoter methylation in malignant gliomas:ready for personalized medicine?[J]. Nat Rev Neurol,2010,6(1):39-51.
[32]Novik K L, Nimmrich I, Genc B, et al. Epigenomics:genome-wide study of methylation phenomena[J]. Curr Issues Mol Biol,2002,4(4):111-128.
[33]Koutsimpelas D, Pongsapich W, Heinrich U, et al. Promoter methylation of MGMT, MLH1 and RASSF1A tumor suppressor genes in head and neck squamous cell carcinoma:pharmacological genome demethylation reduces proliferation of head and neck squamous carcinoma cells[J]. Oncol Rep,2012,27(4):1135-1141.
[34]杨海捷.散发性乳腺癌及乳腺增生组织BRCA1基因启动子区甲基化研究[D].第三军医大学病理学与病理生理学,2004.
[35]Kim J S, Han J, Shim Y M, et al. Aberrant methylation of H-cadherin (CDH13) promoter is associated with tumor progression in primary nonsmall cell lung carcinoma[J]. Cancer, 2005,104(9):1825-1833.
[36]Akiyama Y, Watkins N, Suzuki H, et al. GATA-4 and GATA-5 transcription factor genes and potential downstream antitumor target genes are epigenetically silenced in colorectal and gastric cancer[J]. Mol Cell Biol,2003,23(23):8429-8439.
[37]Suga Y, Miyajima K, Oikawa T, et al. Quantitative p16 and ESR1 methylation in the peripheral blood of patients with non-small cell lung cancer[J]. Oncol Rep, 2008,20(5):1137-1142.
[38]Widschwendter M, Jiang G, Woods C, et al. DNA hypomethylation and ovarian cancer biology[J]. Cancer Res,2004,64(13):4472-4480.
[39]Pappas J J, Toulouse A, Hebert J, et al. Allelic methylation bias of the RARB2 tumor suppressor gene promoter in cancer[J]. Genes Chromosomes Cancer, 2008,47(11):978-993.
[40]Mizuiri H, Yoshida K, Toge T, et al. DNA methylation of genes linked to retinoid signaling in squamous cell carcinoma of the esophagus:DNA methylation of CRBP1 and TIG1 is associated with tumor stage[J]. Cancer Sci,2005,96(9):571-577.
[41]Lin J, Beerm D G. Molecular biology of upper gastrointestinal malignancies[J]. Semin Oncol,2004,31(4):476-486.
[42]舍雅莉.胃癌中RASSF2A和p16基因甲基化及其表达异常的实验研究[D].兰州大学病理与病理生理学,2006.
[43]夏永华.银屑病p16基因启动子CpG岛甲基化状态的研究[D].郑州大学皮肤病学和性病学,2005.
[44]郭晓青.碘染色法结合p16、FHIT基因CpG岛甲基化在食管癌前病变及早期癌中的诊断价值[D].河北医科大学外科学,2005.
[45]姚群峰,康新江,郝巧玲,等.巢式甲基化特异性PCR检测食管癌病人血清中p16基因启动子区过甲基化[J].肿瘤防治研究,2005,32(8)：463-466.
[46]Klump B, Hsieh C J, Holzmann K, et al. Hypermethylation of the CDKN2/p16 promoter during neoplastic progression in Barrett's esophagus[J]. Gastroenterology, 1998,115(6):1381-1386.
[47]Tokugawa T, Sugihara H, Tani T, et al. Modes of silencing of p16 in development of esophageal squamous cell carcinoma[J]. Cancer Res,2002,62(17):4938-4944.
[48]Zhao Y F, Shen S P, Jiang J Y, et al. [Methylation and expression of gene p 16INK4a and RB in breast carcinoma][J]. Zhonghua Bing Li Xue Za Zhi,2010,39(6):377-381.
[49]李华川,陆士新,冯骆.食管癌Rb基因启动子甲基化状态和甲基苄基亚硝胺对其影响的研究[J].中华肿瘤杂志,1998(6)：412.
[50]Peltomaki P. Deficient DNA mismatch repair:a common etiologic factor for colon cancer[J]. Hum Mol Genet,2001,10(7):735-740.
[51]Wu D L, Sui F Y, Jiang X M, et al. Methylation in esophageal carcinogenesis[J]. World J Gastroenterol,2006,12(43):6933-6940.
[52]Wang F, Xie X J, Piao Y S, et al. [Methylation of p16 and hMLH1 genes in esophageal squamous cell carcinoma and reflux esophagitis][J]. Zhonghua Bing Li Xue Za Zhi, 2011,40(8):537-541.
[53]Vasavi M, Ponnala S, Gujjari K, et al. DNA methylation in esophageal diseases including cancer:special reference to hMLH1 gene promoter status[J]. Tumori, 2006,92(2):155-162.
[54]张功员,马春晓,刘秋亮,等.食管癌组织中hMSH2基因启动子区甲基化检测[J].中华肿瘤杂志,2005,27(9)：541-543.
[55]田子强,刘俊峰,李勇,等.食管癌细胞系中MT-3基因CpG岛甲基化的意义[J].实用癌症杂志,2005,20(2)：123-125,137.
[56]Tsao I, Atramentova L A. [Genetic and epigenetic changes of FHIT gene in patients with esophageal cancer][J]. Tsitol Genet,2009,43(6):40-44.
[57]Schildhaus H U, Krockel I, Lippert H, et al. Promoter hypermethylation of p16INK4a, E-cadherin, O6-MGMT, DAPK and FHIT in adenocarcinomas of the esophagus, esophagogastric junction and proximal stomach[J]. Int J Oncol,2005,26(6):1493-1500.
[58]Clement G, Braunschweig R, Pasquier N, et al. Methylation of APC, TIMP3, and TERT: a new predictive marker to distinguish Barrett's oesophagus patients at risk for malignant transformation[J]. J Pathol,2006,208(1):100-107.
[59]WANG C C, MAO W M, LING Z Q. [Expression of DNA methylation of APC in peripheral blood and tumor tissue in patients with esophageal squamous cell carcinoma][J]. Zhonghua Wei Chang Wai Ke Za Zhi,2011,14(9):719-722.
[2]Jemal A, Siegel R, Ward E, et al. Cancer statistics,2009[J]. CA Cancer J Clin, 2009,59(4):225-249.
[3]Guo M, Ren J, Brock M V, et al. Promoter methylation of HIN-1 in the progression to esophageal squamous cancer[J]. Epigenetics,2008,3(6):336-341.
[4]Holliday R. The inheritance of epigenetic defects[J]. Science,1987,238(4824):163-170.
[5]Matias K P. Study of epigenetic changes leads to treatment advances in adult leukemias.Oncol.2003;48:1-4.
[6]张晓梅Dact2在肝癌中的表观遗传学变化及功能研究[D].军医进修学院解放军总医院解放军总医院解放军军医进修学院消化系病,2010.
[7]尹东涛.SOX17在肺癌中的表观遗传改变及功能研究[D].解放军总医院军医进修学院解放军总医院军医进修学院胸外科,2011.
[8]黄庆,郭颖,府伟灵.人类表观基因组计划[J].生命的化学,2004,24(2)：101-103.
[9]Hermann A, Gowher H, Jeltsch A. Biochemistry and biology of mammalian DNA methyltransferases[J]. Cell Mol Life Sci,2004,61(19-20):2571-2587.
[10]王新刚,王笑峰,李福年,等.表观遗传学药物影响乳腺癌细胞系抑癌基因BRCA1及CHD5表达的研究[J].山东大学学报(医学版),2011,49(7)：29-34.
[11]White P S, Thompson P M, Gotoh T, et al. Definition and characterization of a region of 1p36.3 consistently deleted in neuroblastoma [J]. Oncogene,2005,24(16):2684-2694.
[12]Mulero-Navarro S, Esteller M. Chromatin remodeling factor CHD5 is silenced by promoter CpG island hypermethylation in human cancer[J]. Epigenetics,2008,3(4):210-215.
[13]韩琳琳,侯琳,宋金莲.甲硫氨酸合成酶多态性及CHD5基因甲基化与乳腺癌发病的关系[J].中国癌症杂志,2010,20(4)：251-255.
[14]Garber K. Momentum building for human epigenome project[J]. J Natl Cancer Inst, 2006,98(2):84-86.
[15]Holliday R. Epigenetics:an overview[J]. Dev Genet,1994,15(6):453-457.
[16]Smits K M, Cleven A H, Weijenberg M P, et al. Phannacoepigenomics in colorectal cancer:a step forward in predicting prognosis and treatment response[J]. Pharmacogenomics, 2008,9(12):1903-1916.
[17]Jones PA, Laird PW. Cancer epigenetics comes of age[J]. Nat Genet,1999,21(2):163-167.
[18]Paz M F, Fraga M F, Avila S, et al. A systematic profile of DNA methylation in human cancer cell lines[J]. Cancer Res,2003,63(5):1114-1121.
[19]Gama-Sosa M A, Slagel V A, Trewyn R W, et al. The 5-methylcytosine content of DNA from human tumors[J]. Nucleic Acids Res,1983,11(19):6883-6894.
[20]Gonzalez-Zulueta M, Bender C M, Yang A S, et al. Methylation of the 5'CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing[J]. Cancer Res,1995,55(20):4531-4535.
[21]Herman J G, Merlo A, Mao L, et al. Inactivation of the CDKN2/p16/MTS1 gene is frequently associated with aberrant DNA methylation in all common human cancers[J]. Cancer Res, 1995,55(20):4525-4530.
[22]Sudarsanam P, Winston F. The Swi/Snf family nucleosome-remodeling complexes and transcriptional control[J]. Trends Genet,2000,16(8):345-351.
[23]Hall J A, Georgel P T. CHD proteins:a diverse family with strong ties[J]. Biochem Cell Biol, 2007,85(4):463-476.
[24]Bagchi A, Mills A A. The quest for the 1p36 tumor suppressor[J]. Cancer Res, 2008,68(8):2551-2556.
[25]Isidor B, Le Cunff M, Boceno M, et al. Complex constitutional subtelomeric 1p36.3 deletion/duplication in a mentally retarded child with neonatal neuroblastoma[J]. Eur J Med Genet,2008,51(6):679-684.
[26]Okawa E R, Gotoh T, Manne J, et al. Expression and sequence analysis of candidates for the 1p36.31 tumor suppressor gene deleted in neuroblastomas[J]. Oncogene, 2008,27(6):803-810.
[27]Maser R S, Choudhury B, Campbell P J, et al. Chromosomally unstable mouse tumours have genomic alterations similar to diverse human cancers[J]. Nature,2007,447(7147):966-971.
[28]Bagchi A, Papazoglu C, Wu Y, et al. CHD5 is a tumor suppressor at human lp36[J]. Cell, 2007,128(3):459-475.
[29]Zhang C, Li H, Wang Y, et al. Epigenetic inactivation of the tumor suppressor gene RIZ1 in hepatocellular carcinoma involves both DNA methylation and histone modifications [J]. J Hepatol,2010,53(5):889-895.
[30]Smith L T, Lin M, Brena R M, et al. Epigenetic regulation of the tumor suppressor gene TCF21 on 6q23-q24 in lung and head and neck cancer[J]. Proc Natl Acad Sci USA, 2006,103(4):982-987.
[31]Lin J, Beerm D G. Molecular biology of upper gastrointestinal malignancies[J]. Semin Oncol, 2004,31(4):476-486.
[32]Guo M, House M G, Akiyama Y, et al. Hypermethylation of the GATA gene family in esophageal cancer[J]. Int J Cancer,2006,119(9):2078-2083.
[33]Tzao C, Hsu H S, Sun G H, et al. Promoter methylation of the hMLH 1 gene and protein expression of human mutL homolog 1 and human mutS homolog 2 in resected esophageal squamous cell carcinoma[J]. J Thorac Cardiovasc Surg,2005,130(5):1371.
[34]Okawa E R, Gotoh T, Manne J, et al. Expression and sequence analysis of candidates for the 1p36.31 tumor suppressor gene deleted in neuroblastomas[J]. Oncogene, 2008,27(6):803-810.
[1]Jemal A, Siegel R, Ward E, et al. Cancer statistics,2009[J]. CA Cancer J Clin, 2009,59(4):225-249.
[2]Kanavos P. The rising burden of cancer in the developing world[J]. Ann Oncol,2006,17 SuPp18:i15-i23.
[3]Kamangar F, Dores G M, Anderson W F. Patterns of cancer incidence, mortality, and prevalenceacross five continents:defining priorities to reduce cancer disparities in different geographic regions of the world [J]. J Clin Oncol,2006,24 (14):2137-2150.
[4]Pohl H, Welch H G. The role of overdiagnosis and reclassification in the marked increase of esophageal adenocarcinoma incidence[J]. J Natl Cancer Inst,2005,97(2):142-146.
[5]Holliday R. Epigenetics:an overview[J]. Dev Genet,1994,15(6):453-457.
[6]Gama-Sosa M A, Midgett R M, Slagel V A, et al. Tissue-specific differences in DNA methylation in various mammals[J]. Biochim Biophys Acta,1983,740(2):212-219.
[7]Gonzalez-Zulueta M, Bender C M, Yang A S, et al. Methylation of the 5'CpG island of the p16/CDKN2 tumor suppressor gene in normal and transformed human tissues correlates with gene silencing[J]. Cancer Res,1995,55(20):4531-4535.
[8]Herman J G, Latif F, Weng Y, et al. Silencing of the VHL tumor-suppressor gene by DNA methylation in renal carcinoma[J]. Proc Natl Acad Sci USA,1994,91 (21):9700-9704.
[9]Reik W, Lewis A. Co-evolution of X-chromosome inactivation and imprinting in mammals[J]. Nat Rev Genet,2005,6(5):403-410.
[10]Ramsahoye B H, Burnett A K, Taylor C. Restriction endonuclease isoschizomers Ital, BsoFI and Fsp4HI are characterised by differences in their sensitivities to CpG methylation[J]. Nucleic Acids Res,1997,25(16):3196-3198.
[11]Saxonov S, Berg P, Brutlag D L. A genome-wide analysis of CpG dinucleotides in the human genome distinguishes two distinct classes of promoters[J]. Proc Natl Acad Sci U SA,2006,103(5):1412-1417.
[12]Kulis M, Esteller M. DNA methylation and cancer[J]. Adv Genet,2010,70:27-56.
[13]Kato Y, Kaneda M, Hata K, et al. Role of the Dnmt3 family in de novo methylation of imprinted and repetitive sequences during male germ cell development in the mouse[J]. Hum Mol Genet,2007,16(19):2272-2280.
[14]Lin H, Yamada Y, Nguyen S, et al. Suppression of intestinal neoplasia by deletion of Dnmt3b[J]. Mol Cell Biol,2006,26(8):2976-2983.
[15]Deplus R, Brenner C, Burgers W A, et al. Dnmt3L is a transcriptional repressor that recruits histone deacetylase[J]. Nucleic Acids Res,2002,30(17):3831-3838.
[16]Baba Y, Huttenhower C, Nosho K, et al. Epigenomic diversity of colorectal cancer indicated by LINE-1 methylation in a database of 869 tumors[J]. Mol Cancer, 2010,9:125.
[17]白志勇,徐国宾,武淑兰.同位素示踪微量法测定白血病细胞DNA-甲基转移酶活性[J].北京医科大学学报,2000,32(1)：76-78.
[18]郑全辉,张宗玉,童坦君.DNA甲基化在细胞衰老中的作用[J].生命的化学,2003,23(4)：263-265.
[19]Bird A P. The relationship of DNA methylation to cancer[J]. Cancer Surv, 1996,28:87-101.
[20]Galm O, Herman J G, Baylin S B. The fundamental role of epigenetics in hematopoietic malignancies[J]. Blood Rev,2006,20(1):1-13.
[21]Seifert H H, Schmiemann V, Mueller M, et al. In situ detection of global DNA hypomethylation in exfoliative urine cytology of patients with suspected bladder cancer[J]. Exp Mol Pathol,2007,82(3):292-297.
[22]郭晓青,李兆中.DNA甲基化与恶性肿瘤[J].国际消化病杂志,2007,27(1)：57-60.
[23]Wu K, Weng Z, Tao Q, et al. Stage-specific expression of breast cancer-specific gene gamma-synuclein[J]. Cancer Epidemiol Biomarkers Prev,2003,12(9):920-925.
[24]Milicic A, Harrison L A, Goodlad R A, et al. Ectopic expression of P-cadherin correlates with promoter hypomethylation early in colorectal carcinogenesis and enhanced intestinal crypt fission in vivo[J]. Cancer Res,2008,68(19):7760-7768.
[25]Liu H, Liu W, Wu Y, et al. Loss of epigenetic control of synuclein-gamma gene as a molecular indicator of metastasis in a wide range of human cancers[J]. Cancer Res, 2005,65(17):7635-7643.
[26]Roman-Gomez J, Jimenez-Velasco A, Agirre X, et al. Promoter hypomethylation of the LINE-1 retrotransposable elements activates sense/antisense transcription and marks the progression of chronic myeloid leukemia[J]. Oncogene,2005,24(48):7213-7223.
[27]Buchi F, Spinelli E, Masala E, et al. Proteomic analysis identifies differentially expressed proteins in AML1/ETO acute myeloid leukemia cells treated with DNMT inhibitors azacitidine and decitabine[J]. Leuk Res,2012,36(5):607-618.
[28]Saito Y, Kanai Y, Sakamoto M, et al. Overexpression of a splice variant of DNA methyltransferase 3b, DNMT3b4, associated with DNA hypomethylation on pericentromeric satellite regions during human hepatocarcinogenesis[J]. Proc Natl Acad Sci USA,2002,99(15):10060-10065.
[29]Gonzalez-Gomez P, Bello M J, Alonso M E, et al. CpG island methylation status and mutation analysis of the RB1 gene essential promoter region and protein-binding pocket domain in nervous system tumours[J]. Br J Cancer,2003,88(1):109-114.
[30]侯志平,何培元,周伟娜.甲基化与肿瘤的相关性研究[J].承德医学院学报,2010,27(2)：194-196.
[31]Weller M, Stupp R, Reifenberger G, et al. MGMT promoter methylation in malignant gliomas:ready for personalized medicine?[J]. Nat Rev Neurol,2010,6(1):39-51.
[32]Novik K L, Nimmrich I, Genc B, et al. Epigenomics:genome-wide study of methylation phenomena[J]. Curr Issues Mol Biol,2002,4(4):111-128.
[33]Koutsimpelas D, Pongsapich W, Heinrich U, et al. Promoter methylation of MGMT, MLH1 and RASSF1A tumor suppressor genes in head and neck squamous cell carcinoma:pharmacological genome demethylation reduces proliferation of head and neck squamous carcinoma cells[J]. Oncol Rep,2012,27(4):1135-1141.
[34]杨海捷.散发性乳腺癌及乳腺增生组织BRCA1基因启动子区甲基化研究[D].第三军医大学病理学与病理生理学,2004.
[35]Kim J S, Han J, Shim Y M, et al. Aberrant methylation of H-cadherin (CDH13) promoter is associated with tumor progression in primary nonsmall cell lung carcinoma[J]. Cancer, 2005,104(9):1825-1833.
[36]Akiyama Y, Watkins N, Suzuki H, et al. GATA-4 and GATA-5 transcription factor genes and potential downstream antitumor target genes are epigenetically silenced in colorectal and gastric cancer[J]. Mol Cell Biol,2003,23(23):8429-8439.
[37]Suga Y, Miyajima K, Oikawa T, et al. Quantitative p16 and ESR1 methylation in the peripheral blood of patients with non-small cell lung cancer[J]. Oncol Rep, 2008,20(5):1137-1142.
[38]Widschwendter M, Jiang G, Woods C, et al. DNA hypomethylation and ovarian cancer biology[J]. Cancer Res,2004,64(13):4472-4480.
[39]Pappas J J, Toulouse A, Hebert J, et al. Allelic methylation bias of the RARB2 tumor suppressor gene promoter in cancer[J]. Genes Chromosomes Cancer, 2008,47(11):978-993.
[40]Mizuiri H, Yoshida K, Toge T, et al. DNA methylation of genes linked to retinoid signaling in squamous cell carcinoma of the esophagus:DNA methylation of CRBP1 and TIG1 is associated with tumor stage[J]. Cancer Sci,2005,96(9):571-577.
[41]Lin J, Beerm D G. Molecular biology of upper gastrointestinal malignancies[J]. Semin Oncol,2004,31(4):476-486.
[42]舍雅莉.胃癌中RASSF2A和p16基因甲基化及其表达异常的实验研究[D].兰州大学病理与病理生理学,2006.
[43]夏永华.银屑病p16基因启动子CpG岛甲基化状态的研究[D].郑州大学皮肤病学和性病学,2005.
[44]郭晓青.碘染色法结合p16、FHIT基因CpG岛甲基化在食管癌前病变及早期癌中的诊断价值[D].河北医科大学外科学,2005.
[45]姚群峰,康新江,郝巧玲,等.巢式甲基化特异性PCR检测食管癌病人血清中p16基因启动子区过甲基化[J].肿瘤防治研究,2005,32(8)：463-466.
[46]Klump B, Hsieh C J, Holzmann K, et al. Hypermethylation of the CDKN2/p16 promoter during neoplastic progression in Barrett's esophagus[J]. Gastroenterology, 1998,115(6):1381-1386.
[47]Tokugawa T, Sugihara H, Tani T, et al. Modes of silencing of p16 in development of esophageal squamous cell carcinoma[J]. Cancer Res,2002,62(17):4938-4944.
[48]Zhao Y F, Shen S P, Jiang J Y, et al. [Methylation and expression of gene p 16INK4a and RB in breast carcinoma][J]. Zhonghua Bing Li Xue Za Zhi,2010,39(6):377-381.
[49]李华川,陆士新,冯骆.食管癌Rb基因启动子甲基化状态和甲基苄基亚硝胺对其影响的研究[J].中华肿瘤杂志,1998(6)：412.
[50]Peltomaki P. Deficient DNA mismatch repair:a common etiologic factor for colon cancer[J]. Hum Mol Genet,2001,10(7):735-740.
[51]Wu D L, Sui F Y, Jiang X M, et al. Methylation in esophageal carcinogenesis[J]. World J Gastroenterol,2006,12(43):6933-6940.
[52]Wang F, Xie X J, Piao Y S, et al. [Methylation of p16 and hMLH1 genes in esophageal squamous cell carcinoma and reflux esophagitis][J]. Zhonghua Bing Li Xue Za Zhi, 2011,40(8):537-541.
[53]Vasavi M, Ponnala S, Gujjari K, et al. DNA methylation in esophageal diseases including cancer:special reference to hMLH1 gene promoter status[J]. Tumori, 2006,92(2):155-162.
[54]张功员,马春晓,刘秋亮,等.食管癌组织中hMSH2基因启动子区甲基化检测[J].中华肿瘤杂志,2005,27(9)：541-543.
[55]田子强,刘俊峰,李勇,等.食管癌细胞系中MT-3基因CpG岛甲基化的意义[J].实用癌症杂志,2005,20(2)：123-125,137.
[56]Tsao I, Atramentova L A. [Genetic and epigenetic changes of FHIT gene in patients with esophageal cancer][J]. Tsitol Genet,2009,43(6):40-44.
[57]Schildhaus H U, Krockel I, Lippert H, et al. Promoter hypermethylation of p16INK4a, E-cadherin, O6-MGMT, DAPK and FHIT in adenocarcinomas of the esophagus, esophagogastric junction and proximal stomach[J]. Int J Oncol,2005,26(6):1493-1500.
[58]Clement G, Braunschweig R, Pasquier N, et al. Methylation of APC, TIMP3, and TERT: a new predictive marker to distinguish Barrett's oesophagus patients at risk for malignant transformation[J]. J Pathol,2006,208(1):100-107.
[59]WANG C C, MAO W M, LING Z Q. [Expression of DNA methylation of APC in peripheral blood and tumor tissue in patients with esophageal squamous cell carcinoma][J]. Zhonghua Wei Chang Wai Ke Za Zhi,2011,14(9):719-722.