采用基因芯片研究表观遗传修饰在胃癌发生机制中的作用
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摘要
目的研究表观遗传修饰在胃癌发病机制中的作用。方法收集我科2007.10～2008.01病理确诊的8例胃癌患者肿瘤组织和正常胃粘膜组织，采用染色质免疫共沉淀联合芯片技术(ChIP-chip)在细胞全基因组范围内对两种组织细胞内组蛋白H3K27位点三甲基化水平进行高通量的检测，用染色质免疫共沉淀-实时定量聚合酶链反应(ChIP-qPCR)验证芯片检测结果，采用荧光定量反转录聚合酶链反应(qRT-PCR)检测两种组织细胞H3K27me3水平存在显著差异的目的基因mRNA表达水平，最后用甲基化DNA免疫沉淀-定量PCR(MeDIP-qPCR)方法检测目的基因启动子区域DNA甲基化水平。结果和正常胃粘膜组织细胞相比较，胃癌肿瘤细胞全基因组范围共筛选出227个基因组蛋白H3K27me3水平存在显著差异，其中有67个基因显示组蛋白H3K27三甲基化水平增高，160个基因组蛋白H3K27三甲基化水平降低；ChIP-qPCR技术检测两种组织细胞挑选基因组蛋白H3K27三甲基化水平的差异与CpG岛芯片检测结果一致，两种组织细胞挑选基因的mRNA表达水平和DNA甲基化水平均存在显著差异。结论与正常胃粘膜细胞相比，胃癌肿瘤细胞多个基因组蛋白H3K27三甲基化水平存在显著改变；基因组蛋白H3K27三甲基化对调控基因表达发挥沉默效应；挑选基因DNA甲基化水平亦存在明显变化，挑选基因DNA甲基化和组蛋白H3K27me3两种表观修饰方式存在相互作用，DNA甲基化修饰作用可能存在优势；表观遗传修饰在胃癌发病机制中发挥重要作用。ChIP-chip技术有利于进一步揭示胃癌发生的分子机制，发现新的治疗靶点。
Objective To Study the effect of epigenetic gene modification on gastric cancer moleculor mechanism. Methods 8 cases including tumor tissue and gastric mucosa tissue in gastric cancer patients were collected from 10.2007 to 01.2008. For the first time chromatin immunoprecipitation linked to microarrays (ChIP-chip) was adopted to profile the variations in histone H3K27me3 of Genome-wide in tumor tissue and gastric mucosa tissue. ChIP-qPCR was used to validate the microrray results. mRNA Expression analysis of 3 selected genes by qRT-PCR was performed to confirm the correlations between H3K27me3 and gene expression. Finally, DNA methylation of five selected genes was detected with MeDIP-qPCR. Results 227(67 increased and 160 decreased) gene displayed significant histone H3K27me3 difference were found in gastric cancer cell compared with gastric mucosa tissue. The results of ChIP-qPCR were coincided well with ChIP-chip. There was marked difference between tumor tissue and gastric mucosa tissue about mRNA expression of selected genens, so is DNA methylation. Conclusions Compared with gastric mucosa tissue, there are significant changes in tumor tissue about histone H3K27me3 profiling and DNA methylation level. It is feasble that ChIP-chip technique detects histone H3K27me3 level. Histone H3K27me3 probably bring gene expression about "silence effect". There is interaction between histone H3K27me3 and DNA methylation. But DNA methylation is probably more important than H3K27me3. Epigenetic gene modification probably plays an important role in gastric cancer moleculor mechanism. These novel candidate genes may be become potential biomarkers or future therapeutic targets. The ChIP-chip technique will help further reveal gastric cancer molecular mechanisms and discover new therapeutic targets.
Objective To Study the effect of epigenetic gene modification on gastric cancer moleculor mechanism. Methods 8 cases including tumor tissue and gastric mucosa tissue in gastric cancer patients were collected from 10.2007 to 01.2008. For the first time chromatin immunoprecipitation linked to microarrays (ChIP-chip) was adopted to profile the variations in histone H3K27me3 of Genome-wide in tumor tissue and gastric mucosa tissue. ChIP-qPCR was used to validate the microrray results. mRNA Expression analysis of 3 selected genes by qRT-PCR was performed to confirm the correlations between H3K27me3 and gene expression. Finally, DNA methylation of five selected genes was detected with MeDIP-qPCR. Results 227(67 increased and 160 decreased) gene displayed significant histone H3K27me3 difference were found in gastric cancer cell compared with gastric mucosa tissue. The results of ChIP-qPCR were coincided well with ChIP-chip. There was marked difference between tumor tissue and gastric mucosa tissue about mRNA expression of selected genens, so is DNA methylation. Conclusions Compared with gastric mucosa tissue, there are significant changes in tumor tissue about histone H3K27me3 profiling and DNA methylation level. It is feasble that ChIP-chip technique detects histone H3K27me3 level. Histone H3K27me3 probably bring gene expression about "silence effect". There is interaction between histone H3K27me3 and DNA methylation. But DNA methylation is probably more important than H3K27me3. Epigenetic gene modification probably plays an important role in gastric cancer moleculor mechanism. These novel candidate genes may be become potential biomarkers or future therapeutic targets. The ChIP-chip technique will help further reveal gastric cancer molecular mechanisms and discover new therapeutic targets.
引文
1. Lichtenstein P, Holm NV, Verkasalo PK, etal. Environmental and heritable factors in the causation of cancer-analyses of cohorts of twins from Sweden, Denmark, and Finland [J]. N Engl J Med, 2000, 343(2):78-85.
2. Wolffe AP, Matzke MA. Epigenetics: regulation through repression [J]. Science,1999,286 (5439): 481-486.
3. JenuweinT, Allis CD.Translating the histone code [J]. Science, 2001, 293(5532):1074-1080.
4. Yasui W, Oue N, Ono S, et al. Histone acetylation and gastro-intestinal carcinogenesis[J]. Ann N Y Acad Sci, 2003, 983(3): 220-231.
5. Kim J H, Choi Y K. Downregulation of gelsolin and retinoic acid receptor expression in gastric cancer tissues through histone deacetylase 1[J]. J Gastroenterol Hepatol,2004, 19(2): 218-224.
6. Ono S, Oue N, Kuniyasu H. Acetylated histone H4 is reduced in human gastric adenomas and carcinomas [J]. J Exp Clin Cancer Res, 2002, 21(3): 377-382.
7. Zhang Y, Reinberg D. Transcription regulation by histone meth-ylation: interplay between different covalent modifications of the core histone tails [J]. Genes Dev, 2001, 15(18): 2343-2360
8. Wysocka J, Swigut T. WDR5 associates with histone H3 meth-ylated at K4 and is essential for H3K4 methylation and verte-brate development [J]. Cell, 2005, 121(6): 859-872.
9. Hake S B, Xiao A, Aills C D. Linking the epigenetic language of covalent histone modifications to cancer [J]. British J Cancer, 2004, 90(4): 761-769.
10. Kondo Y, Shen L, Issa JP. Critical role of histone methylation in tumor suppressor gene silencing in colorectal cancer [J]. Mol Cell Biol, 2003, 23(1): 206-215.
11. Robertson KD. DNA methylation and human disease [J]. Nat Rev Genet, 2005, 6(8):597-610.
12. Sakuma K, Chong JM, Sudo M, et al. High-density methylation of pl4ARF and p16INK4A in Epstein-Barrvirus-associated gastric car-cinoma. Int JCancer, 2004,112(2): 273-278.
13. Lee YY, Kang SH, Seo JY, et al. Alterations of pl6INK4A and pl5INK4B genes in gastric carcinomas. Cancer, 1997, 80(10): 1889-1896.
14. KanyamaY, HiroshiK, NakayamaH, et al. Detection ofP16 promoter hypermethylation in serum of gastric cancer patients. Cancer Sci, 2003, 94(5): 418-420.
15. EbertMP, Yu J, Hoffmann J, et al. Loss of beta-catenin expression in metastatic gastric cancer. J Clin Oncol. 2003, 21(9): 1708-1714.
16. Byun DS, LeeMG, ChaeKS, et al. Frequent epigenetic inactivation of RASSF1A by aberrant promoter hypermethylation in human gastric adenocarcinoma. Cancer Res,2001, 61 (19): 7034-7038.
17. Fujimoto J, Yasui W, Tahara H, et al. DNA hypermethylation at the pS2 promoter region is associatedwith early stage of stomach carcino-genesis. Cancer Lett, 2000,149(1-2): 125-134.
18. Cunningham JM, Christensen ER, Tester DJ, et al. Hypermethylation of the hMLH1 promoter in colon cancerwith microsatellite instabili-ty. Cancer Res, 1998, 58 (15):3455-3460.
19. Rhee I, JairKW, Yen RW, et al. CpG methylation ismaintained in human cancer cells lacking DNMT1. Nature, 2000, 404 (6781): 1003-1007-20. Toyota M, Ahuja N, OheToyota M, etal. CpG island methylator phe-notype in colorectal cancer. Proc Natl Acad Sci USA, 1999, 96(15): 8681-8686.
21. Herman JG, Graff JR, Myohanen S, et al. Methylation-specific PCR: a nov-el PCR assay for methylation status of CpG islands [J]. Proc Natl AcadSci USA, 1996, 93(18):9821-6.
22. Mukesh V, Sudhir S. Epigenetics in cancer: implications for early detection and prevention [J]. Lancet Oncol, 2002, 3(12):755-763.
23. Kenneth P, Nephewa, Tim Hui-Ming H. Epigenetic gene silencing in cancer initiation and progression [J]. Cancer Lett, 2003, 190(2):125-133.
24．陈峻青。近半个世纪胃癌外科治疗的变革与现状。中国实用外科杂志。2007，27(7)：501-503。
1. Shannon M F, Rao S. Of Chips and ChIPs. Science, 2002, 296(5568): 666-669.
2. Bannister AJ, Kouzarides T. Histone methylation: recognizing the methyl mark.Methods Enzymol 2004; 376:269-288.
3. Jenuwein T, Allis CD. Translating the histone code. Science, 2001; 293: 1074-1080.
4. WeinbergM S, Villeneuve LM, EhsaniA, et a.l The antisense strand of small interfering RNAs directs histone methylation and transcriptional gene silencing in human cells [J].RNA, 2006, 12(2): 256-262.
5. Wang H, Wang L, Erdjument-Bromage H, Vidal M, Tempst P, Jones RS, Zhang Y.Role of histone H2A ubiq-uitination in Polycomb silencing. Nature, 2004, 431(7010):873-878.
6. Weinmann AS, Farnham PJ, Identification of unknown target genes of human transcription factors using chromatin immunoprecipitation. Methods, 2002,26(l):37-44
7. Hunter T. Signaling 2000 and beyond. Cell, 2000,100(1): 113-140
8. Schlessinger J. Cell sigaling by receptor tyrosine kinases. Cell, 2000,103(2):211-236.
9. Thiebault K, Mazelin L, Pays L, 'Llambi F, Joly MO, Saurin JC, et al. The netrin-1 receptors UNC5H are putative tumor suppressors controlling cell death commitment. Proc Natl Acad Sci USA, 2003; 100(7): 4173-4178.
10. Llambi F, Lourengo FC, Gozuacik D, Guix C, Pays L, Del Rio G,et al.The dependence receptor UNC5H2 mediates apoptosis through DAP-kinase. EMBO J, 2005; 24(6):1192-1201.
11. Tanikawa C, Matsuda K, Fukuda S, Nakamura Y, Arakawa H. p53RDLl regulates p53- dependent apoptosis. Nat Cell Biol ,2003; 5(3): 216-223.
12. Robert V, Hideaki N. Matrix metalloproteinases and tissue inhibi-tors of metalloproteinases. Circul Res, 2003, 92(8):827-839.
13. Giraudo E, Inoue M, Hanahan D. An amino-bisphosphonate tar-gets MMP-9-expressing macrophages and angiogenesis to impaircervical carcinogenesis. J Clin Invest, 2004, 114(5): 623-633.
14. Kong Y, Poon R, Nadesan P, et al. Matrix metalloproteinase ac-tivity modulates tumor size, cell motility, and cell invasiveness inmurine aggressive fibromatosis. Cancer Res,2004, 64(16): 5795-5803.
15. Hotary K, Li X Y, Allen E, et al. A cancer cell metalloprotease triad regulates the basement membrane transmigration program. Genes Dev. 2006; 20(19): 2673-2686.
16. El-Amraoui A, Schonn JS, Kiissel-Andermann P, et al. MyRIP, a novel Rab effector, enables myosin Vila recruitment to retinal melanosomes. EMBO Rep. 2002; 3(5): 463-470.
17. Morin P J, Sparks A B, Korinek V, et al. Activation of beta-catenin-Tcf signaling in colon cancer by mutations in beta-catenin or APC. Science, 1997; 275(5307):1787-1790.
18. Salvan A, Thomaseth K, Bortot P, et al. Use of a toxicokinetic model in the analysis of cancer mortality in relation to theestimated absorbed dose of dioxin. Sci Total Environ,2001, 274(1-3): 21-35.
19. Cohen M M. The hedgehog-signaling network [J]. Am J Med Genet, 2003, 123A (1):5-28.
20. Berman D M, Karhadkar S S, Maitra A, et al. Widespread requirement for Hedgehog ligand stimulation in growth of digestive tract tumors [J]. Nature, 2003, 425(6960):846-851.
21. Mochizuki S, ShimodaM, ShiomiT, etal. ADAM28 is activated by MMP-7 (matrilysin-1) and cleaves insulin- like growth factor binding protem-3[J].Biochem Biophys Res Commun, 2004, 315(1): 79-84
22. Ohtsuka T, Shiomi T, ShimodaM, et al. ADAM28 is overexpressed in human non-small cell lung carcinomas and correlates with cell proliferation and lymph node metastasis[J]. Int J Cancer, 2006, 118(2): 263-273.
23. Muller HM, Widschwendter A, Fiegl H, et al. DNA methylation in serum of breast cancer patients: an independent prognostic marker. Cancer Res, 2003, 63(22):7641-7645.
24. Leong ML, Maiyar AC, Kim B, et al. Expression of the serum and glucocorticoid-inducible protein kinase, sgk, is a cell survival responseto multiple types of envionmental stress stimuli in mammary epithelial cells [J]. J Biol Chem, 2003,278(8): 5871-5882.
25. Kas K, Voz ML, Roijer E, et al. Promoter swapping between the genes for a novel zinc finger protein and beta-catenin in pleomorphic adenomas with t(3;8)(p21;ql2) translations. Nat Genet, 1997,15(2):170-174.
26. Ringrose L, Paro R. Epigenetic regulation of cellular memory by the Polycomb and Trithorax group proteins. Annu Rev Genet, 2004, 38(3): 413-443.
27. Wang H, Wang L, Erdjument-Bromage H, et al. Tempst P, Jones RS, Zhang Y. Role of histone H2A ubiquitination in Polycomb silencing. Nature, 2004, 431(7010):873-878.
1. Shannon M F, Rao S. Of Chips and ChIPs.Science, 2002, 296(5568):666-669.
2．严明，陈万涛，李军等。口腔鳞状细胞癌转移相关基因实时定量PCR的验证。北京口腔医学，2007，15(3)：121-125
3. Higuchi R, Fockler C, Dollinger G, et al. Kinetic PCR analysis: real-time monitoring of DNA amplification reactions [J]. Biotechnology, 1993; 11(9): 1026-30.
4. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2~(-△△CT) method. Methods 2001; 25(4): 402-408.
5. Ueno H, Yoshida K, Hirai T Quantitative detection of carcinoembryonic antigen messenger RNA in the peritoneal cavity of gastric cancer patients by real-time quantitative reverse transcription polymerase chain reaction [J]. Anticancer Res, 2003,23(2C): 1701-8.
6. Hotary K, Li X Y, Allen E, et al. A cancer cell metalloprotease triad regulates the basement membrane transmigration program. Genes Dev. 2006 October 1; 20(19): 2673-2686.
7. Toftgard R. Hedgehog signaling in cancer [J]. Cell Mollife Sci, 2000, 57 (12):1720-1731.
8. Giacinti C, Giordano A. RB and cell cycle progression. Oncogene, 2006; 25(38):5220-5227.
9. Fuego J, Gomez MC, Yung WK, et al. Suppression of hman glioma growth byadnovirus emdiated Rb gene transfer. Neurology, 1998, 50(5): 1 307-1 315.
10. Xu HJ, Kuzumaki N, Kawakami Y, et al. Altered retinoblastoma protein expression in non small cell lung cancer: its synergistic effectswith altered as and p 53 protein status on prognosis. Cancer, 1997, 79(11):1329-1337.
1. Liss B. Improved quantitative real-time RT-PCR for expression profiling of individual cells. Nucleic Acids Res, 2002, 30(17): e89.
2. Glockner S, Lehmann U, Wilke N, et al. Detection of gene amplification in intraductal and infiltrating breast cancer by laser-assisted microdissection and quantitative real-time PCR.Pathobiology, 2000,68(45): 173-179.
3. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2~(-△△CT) method. Methods 2001; 25(4): 402-8.
4. Ringrose L, Paro R. Epigenetic regulation of cellular memory by the Polycomb and Trithorax group proteins. Annu Rev Genet, 2004, 38: 413-443.
5. Wang H, Wang L, Erdjument-Bromage H, et al. Tempst P, Jones RS, Zhang Y. Role of histone H2A ubiquitination in Polycomb silencing. Nature, 2004, 431(7010): 873-878.
6. Dahmane N, Sanehez P, Gitton Y, et al. The Sonic Hedgehog-Glipathway regulates dorsal brain growth and tunmorigenesis [J]. Development, 2001,128(24): 5201-5212.
7. Giacinti C, Giordano A. RB and cell cycle progression. Oncogene, 2006; 25(38):5220-5227.
1. Bergqvist D, Lindholm C, Nelzen O, et al. Chronic leg ulcers: the impact of venous disease [J], J Vasc Surg, 1999, 29(8):752-755.
2. Kondo Y, Shen L, Issa J P. Critical role of histone methylation in tumor suppressor gene silencing in colorectal cancer. Mol Cell Biol, 2003, 23(1): 206-215.
3．钟克力，张丽，戴勇等，采用染色质免疫沉淀联合芯片技术分析胃癌全基因组蛋白H3K27三甲基化水平，临床肿瘤杂志，2009．14(1)：15-19。
4. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2~(-△△CT) method. Methods 2001; 25(4): 402-8.
5. Reik W, Dean W. DNA methylation and mammalian epigenetics, Electrophoresis, 2001,22(9): 2838-2843.
6．王志伟，陈玉英，易静等。乳腺癌P16基因甲基化及其临床病理学特征的关系。中华实验外科杂志，2002，19(2)：81。
7．戴观荣，景洪标，邓鉴文。胃癌中脆性组氨酸三联体基因的甲基化和表达及其意义。中华实验外科杂志，2006，23(11)：594-596。
8. Wu J, Wang SH, Potter D, et al. Diverse histone modifications on histone 3 lysine 9 and their relation to DNA methylation in specifying gene silencing. BMC Genomics, 2007,8(3):131.
9. Irvine RA, Lin IG, Hsieh CL. DNA methylation has a local effect on transcription and histone acetylation. Mol Cell Biol, 2002, 22(7):6689-6696.
10. Okitsu CY, Hsieh CL. DNA methylation dictates histone H3K4 methylation. Mol Cell Biol, 2007, 27(8): 2746- 2757.
11．李宇，司聚同，王永潮等。RB、P53、c-myc和Ki-ras基因异常改变在大肠癌发生、 发展中的意义。中华实验外科杂志，1997，14(5)：152。
12. Tamaru H, Selker EU. A histone H3 methyltransferase controls DNA methylation in Neurospora crassa[J]. Nature, 2001, 414(6861):277-83.
13．宋跃，赵青，林海江等。RB基因在结直肠癌中的重排与扩增。白求恩医科大学学报，1998．24(4)：344-346。
14. Kim SK, Hebrok M. Intercellular signals regulating pancreas development and function. Genes Dev 2001; 15:111-127.
1. Wolffe AP, Matzke MA. Epigenetics: regulation through repression [J]. Science,1999,286 (5439): 481-486.
2. Feinberg AP, Vogelstein B. Hypomethylation distinguishes genes of some human cancers from their normal counterparts [J]. Nature, 1983,301 (5895): 89-92.
3． 腾玥，戴冬秋。胃癌表遗传学的研究进展[J]。世界华人消化杂志，2005，13(19)：2289-2293。
4. Sakuma K, Chong JM, Sudo M, et al. High-density methylation of pl4ARF and pl6INK4A in Epstein-Barrvirus-associated gastric car-cinoma. Int JCancer, 2004,112(2): 273-278.
5. Lee YY, Kang SH, Seo JY, et al. Alterations of pl6INK4A and pl5INK4B genes in gastric carcinomas. Cancer, 1997, 80(10): 1889-1896.
6. KanyamaY, HiroshiK, NakayamaH, et al. Detection ofP16 promoter hypermethylation in serum of gastric cancer patients. Cancer Sci, 2003, 94(5): 418-420.
7. EbertMP, Yu J, Hoffmann J, et al. Loss of beta-catenin expression in metastatic gastric cancer. J Clin Oncol. 2003, 21(9): 1708-1714.
8. Byun DS, LeeMG, ChaeKS, et al. Frequent epigenetic inactivation of RASSF1A by aberrant promoter hypermethylation in human gastric adenocarcinoma. Cancer Res,2001, 61 (19): 7034-7038.
9. Fujimoto J, Yasui W, Tahara H, et al. DNA hypermethylation at the pS2 promoter region is associatedwith early stage of stomach carcino-genesis. Cancer Lett, 2000,149(1-2): 125-134.
10. Cunningham JM, Christensen ER, Tester DJ, et al. Hypermethylation of the hMLHl promoter in colon cancerwith microsatellite instabili-ty. Cancer Res, 1998, 58 (15):3455-3460.
11. Rhee I, JairKW, Yen RW, et al. CpG methylation ismaintained in human cancer cells lacking DNMT1. Nature, 2000, 404 (6781): 1003-1007-
12. Xie S, Wang Z, Okano M, et al. Cloning expression and chromosome locations of the human DNMT3 gene family. Gene, 1999, 236(1): 87-95.
13. Toyota M, Ahuja N, OheToyota M, etal. CpG island methylator phe-notype in colorectal cancer. Proc Natl Acad Sci USA, 1999, 96(15): 8681-8686.
14. Mukesh V, Sudhir S. Epigenetics in cancer: implications for early detection and prevention [J]. Lancet Oncol, 2002, 3(7):755-763.
15. Kenneth P, Nephewa, Tim Hui-Ming H. Epigenetic gene silencing in cancer initiation and progression [J]. Cancer Lett, 2003, 190(7): 125-133.
16. Yasui W, Oue N, Ono S, et al. Histone acetylation and gastro-intestinal carcinogenesis [J]. Ann N Y Acad Sci, 2003, 983(3): 220-231.
17. Kim J H, Choi Y K. Downregulation of gelsolin and retinoic acid receptor expression in gastric cancer tissues through histone deacetylase 1[J]. J Gastroenterol Hepatol,2004, 19(2):218-224.
18. Ono S, Oue N, Kuniyasu H. Acetylated histone H4 is reduced in human gastric adenomas and carcinomas [J]. J Exp Clin Cancer Res, 2002, 21(3): 377-382.
19. Zhang Y, Reinberg D. Transcription regulation by histone meth-ylation: interplay between different covalent modifications of the core histone tails [J]. Genes Dev, 2001,15(18): 2343-2360
20. Wysocka J, Swigut T. WDR5 associates with histone H3 meth-ylated at K4 and is essential for H3K4 methylation and verte-brate development [J]. Cell, 2005, 121(6):859-872.
21. Hake S B, Xiao A, Aills C D. Linking the epigenetic language of covalent histone modifications to cancer [J]. British J Cancer, 2004, 90(4): 761-769.
22. Kondo Y, Shen L, Issa JP. Critical role of histone methylation in tumor suppressor gene silencing in colorectal cancer [J]. Mol Cell Biol, 2003, 23(1): 206-215.
23. Cameron E E, Bachman K E, Myohanen S, et al. Synergy of demethylation and histone deacetylase inhibition in there: expression of genes silenced in cancer [J]. Nat Genet,1999, 21(1): 103-107.
24. Tamaru H, Selker EU.A histone H3 methyltransferase controls DNA methylation in Neurospora crassa [J]. Nature, 2001, 414(6861): 277-83.
25. Jenuwein T, Allis CD.Translating the histone code [J]. Science, 2001, 293(5532):1074-80.
2. Wolffe AP, Matzke MA. Epigenetics: regulation through repression [J]. Science,1999,286 (5439): 481-486.
3. JenuweinT, Allis CD.Translating the histone code [J]. Science, 2001, 293(5532):1074-1080.
4. Yasui W, Oue N, Ono S, et al. Histone acetylation and gastro-intestinal carcinogenesis[J]. Ann N Y Acad Sci, 2003, 983(3): 220-231.
5. Kim J H, Choi Y K. Downregulation of gelsolin and retinoic acid receptor expression in gastric cancer tissues through histone deacetylase 1[J]. J Gastroenterol Hepatol,2004, 19(2): 218-224.
6. Ono S, Oue N, Kuniyasu H. Acetylated histone H4 is reduced in human gastric adenomas and carcinomas [J]. J Exp Clin Cancer Res, 2002, 21(3): 377-382.
7. Zhang Y, Reinberg D. Transcription regulation by histone meth-ylation: interplay between different covalent modifications of the core histone tails [J]. Genes Dev, 2001, 15(18): 2343-2360
8. Wysocka J, Swigut T. WDR5 associates with histone H3 meth-ylated at K4 and is essential for H3K4 methylation and verte-brate development [J]. Cell, 2005, 121(6): 859-872.
9. Hake S B, Xiao A, Aills C D. Linking the epigenetic language of covalent histone modifications to cancer [J]. British J Cancer, 2004, 90(4): 761-769.
10. Kondo Y, Shen L, Issa JP. Critical role of histone methylation in tumor suppressor gene silencing in colorectal cancer [J]. Mol Cell Biol, 2003, 23(1): 206-215.
11. Robertson KD. DNA methylation and human disease [J]. Nat Rev Genet, 2005, 6(8):597-610.
12. Sakuma K, Chong JM, Sudo M, et al. High-density methylation of pl4ARF and p16INK4A in Epstein-Barrvirus-associated gastric car-cinoma. Int JCancer, 2004,112(2): 273-278.
13. Lee YY, Kang SH, Seo JY, et al. Alterations of pl6INK4A and pl5INK4B genes in gastric carcinomas. Cancer, 1997, 80(10): 1889-1896.
14. KanyamaY, HiroshiK, NakayamaH, et al. Detection ofP16 promoter hypermethylation in serum of gastric cancer patients. Cancer Sci, 2003, 94(5): 418-420.
15. EbertMP, Yu J, Hoffmann J, et al. Loss of beta-catenin expression in metastatic gastric cancer. J Clin Oncol. 2003, 21(9): 1708-1714.
16. Byun DS, LeeMG, ChaeKS, et al. Frequent epigenetic inactivation of RASSF1A by aberrant promoter hypermethylation in human gastric adenocarcinoma. Cancer Res,2001, 61 (19): 7034-7038.
17. Fujimoto J, Yasui W, Tahara H, et al. DNA hypermethylation at the pS2 promoter region is associatedwith early stage of stomach carcino-genesis. Cancer Lett, 2000,149(1-2): 125-134.
18. Cunningham JM, Christensen ER, Tester DJ, et al. Hypermethylation of the hMLH1 promoter in colon cancerwith microsatellite instabili-ty. Cancer Res, 1998, 58 (15):3455-3460.
19. Rhee I, JairKW, Yen RW, et al. CpG methylation ismaintained in human cancer cells lacking DNMT1. Nature, 2000, 404 (6781): 1003-1007-20. Toyota M, Ahuja N, OheToyota M, etal. CpG island methylator phe-notype in colorectal cancer. Proc Natl Acad Sci USA, 1999, 96(15): 8681-8686.
21. Herman JG, Graff JR, Myohanen S, et al. Methylation-specific PCR: a nov-el PCR assay for methylation status of CpG islands [J]. Proc Natl AcadSci USA, 1996, 93(18):9821-6.
22. Mukesh V, Sudhir S. Epigenetics in cancer: implications for early detection and prevention [J]. Lancet Oncol, 2002, 3(12):755-763.
23. Kenneth P, Nephewa, Tim Hui-Ming H. Epigenetic gene silencing in cancer initiation and progression [J]. Cancer Lett, 2003, 190(2):125-133.
24．陈峻青。近半个世纪胃癌外科治疗的变革与现状。中国实用外科杂志。2007，27(7)：501-503。
1. Shannon M F, Rao S. Of Chips and ChIPs. Science, 2002, 296(5568): 666-669.
2. Bannister AJ, Kouzarides T. Histone methylation: recognizing the methyl mark.Methods Enzymol 2004; 376:269-288.
3. Jenuwein T, Allis CD. Translating the histone code. Science, 2001; 293: 1074-1080.
4. WeinbergM S, Villeneuve LM, EhsaniA, et a.l The antisense strand of small interfering RNAs directs histone methylation and transcriptional gene silencing in human cells [J].RNA, 2006, 12(2): 256-262.
5. Wang H, Wang L, Erdjument-Bromage H, Vidal M, Tempst P, Jones RS, Zhang Y.Role of histone H2A ubiq-uitination in Polycomb silencing. Nature, 2004, 431(7010):873-878.
6. Weinmann AS, Farnham PJ, Identification of unknown target genes of human transcription factors using chromatin immunoprecipitation. Methods, 2002,26(l):37-44
7. Hunter T. Signaling 2000 and beyond. Cell, 2000,100(1): 113-140
8. Schlessinger J. Cell sigaling by receptor tyrosine kinases. Cell, 2000,103(2):211-236.
9. Thiebault K, Mazelin L, Pays L, 'Llambi F, Joly MO, Saurin JC, et al. The netrin-1 receptors UNC5H are putative tumor suppressors controlling cell death commitment. Proc Natl Acad Sci USA, 2003; 100(7): 4173-4178.
10. Llambi F, Lourengo FC, Gozuacik D, Guix C, Pays L, Del Rio G,et al.The dependence receptor UNC5H2 mediates apoptosis through DAP-kinase. EMBO J, 2005; 24(6):1192-1201.
11. Tanikawa C, Matsuda K, Fukuda S, Nakamura Y, Arakawa H. p53RDLl regulates p53- dependent apoptosis. Nat Cell Biol ,2003; 5(3): 216-223.
12. Robert V, Hideaki N. Matrix metalloproteinases and tissue inhibi-tors of metalloproteinases. Circul Res, 2003, 92(8):827-839.
13. Giraudo E, Inoue M, Hanahan D. An amino-bisphosphonate tar-gets MMP-9-expressing macrophages and angiogenesis to impaircervical carcinogenesis. J Clin Invest, 2004, 114(5): 623-633.
14. Kong Y, Poon R, Nadesan P, et al. Matrix metalloproteinase ac-tivity modulates tumor size, cell motility, and cell invasiveness inmurine aggressive fibromatosis. Cancer Res,2004, 64(16): 5795-5803.
15. Hotary K, Li X Y, Allen E, et al. A cancer cell metalloprotease triad regulates the basement membrane transmigration program. Genes Dev. 2006; 20(19): 2673-2686.
16. El-Amraoui A, Schonn JS, Kiissel-Andermann P, et al. MyRIP, a novel Rab effector, enables myosin Vila recruitment to retinal melanosomes. EMBO Rep. 2002; 3(5): 463-470.
17. Morin P J, Sparks A B, Korinek V, et al. Activation of beta-catenin-Tcf signaling in colon cancer by mutations in beta-catenin or APC. Science, 1997; 275(5307):1787-1790.
18. Salvan A, Thomaseth K, Bortot P, et al. Use of a toxicokinetic model in the analysis of cancer mortality in relation to theestimated absorbed dose of dioxin. Sci Total Environ,2001, 274(1-3): 21-35.
19. Cohen M M. The hedgehog-signaling network [J]. Am J Med Genet, 2003, 123A (1):5-28.
20. Berman D M, Karhadkar S S, Maitra A, et al. Widespread requirement for Hedgehog ligand stimulation in growth of digestive tract tumors [J]. Nature, 2003, 425(6960):846-851.
21. Mochizuki S, ShimodaM, ShiomiT, etal. ADAM28 is activated by MMP-7 (matrilysin-1) and cleaves insulin- like growth factor binding protem-3[J].Biochem Biophys Res Commun, 2004, 315(1): 79-84
22. Ohtsuka T, Shiomi T, ShimodaM, et al. ADAM28 is overexpressed in human non-small cell lung carcinomas and correlates with cell proliferation and lymph node metastasis[J]. Int J Cancer, 2006, 118(2): 263-273.
23. Muller HM, Widschwendter A, Fiegl H, et al. DNA methylation in serum of breast cancer patients: an independent prognostic marker. Cancer Res, 2003, 63(22):7641-7645.
24. Leong ML, Maiyar AC, Kim B, et al. Expression of the serum and glucocorticoid-inducible protein kinase, sgk, is a cell survival responseto multiple types of envionmental stress stimuli in mammary epithelial cells [J]. J Biol Chem, 2003,278(8): 5871-5882.
25. Kas K, Voz ML, Roijer E, et al. Promoter swapping between the genes for a novel zinc finger protein and beta-catenin in pleomorphic adenomas with t(3;8)(p21;ql2) translations. Nat Genet, 1997,15(2):170-174.
26. Ringrose L, Paro R. Epigenetic regulation of cellular memory by the Polycomb and Trithorax group proteins. Annu Rev Genet, 2004, 38(3): 413-443.
27. Wang H, Wang L, Erdjument-Bromage H, et al. Tempst P, Jones RS, Zhang Y. Role of histone H2A ubiquitination in Polycomb silencing. Nature, 2004, 431(7010):873-878.
1. Shannon M F, Rao S. Of Chips and ChIPs.Science, 2002, 296(5568):666-669.
2．严明，陈万涛，李军等。口腔鳞状细胞癌转移相关基因实时定量PCR的验证。北京口腔医学，2007，15(3)：121-125
3. Higuchi R, Fockler C, Dollinger G, et al. Kinetic PCR analysis: real-time monitoring of DNA amplification reactions [J]. Biotechnology, 1993; 11(9): 1026-30.
4. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2~(-△△CT) method. Methods 2001; 25(4): 402-408.
5. Ueno H, Yoshida K, Hirai T Quantitative detection of carcinoembryonic antigen messenger RNA in the peritoneal cavity of gastric cancer patients by real-time quantitative reverse transcription polymerase chain reaction [J]. Anticancer Res, 2003,23(2C): 1701-8.
6. Hotary K, Li X Y, Allen E, et al. A cancer cell metalloprotease triad regulates the basement membrane transmigration program. Genes Dev. 2006 October 1; 20(19): 2673-2686.
7. Toftgard R. Hedgehog signaling in cancer [J]. Cell Mollife Sci, 2000, 57 (12):1720-1731.
8. Giacinti C, Giordano A. RB and cell cycle progression. Oncogene, 2006; 25(38):5220-5227.
9. Fuego J, Gomez MC, Yung WK, et al. Suppression of hman glioma growth byadnovirus emdiated Rb gene transfer. Neurology, 1998, 50(5): 1 307-1 315.
10. Xu HJ, Kuzumaki N, Kawakami Y, et al. Altered retinoblastoma protein expression in non small cell lung cancer: its synergistic effectswith altered as and p 53 protein status on prognosis. Cancer, 1997, 79(11):1329-1337.
1. Liss B. Improved quantitative real-time RT-PCR for expression profiling of individual cells. Nucleic Acids Res, 2002, 30(17): e89.
2. Glockner S, Lehmann U, Wilke N, et al. Detection of gene amplification in intraductal and infiltrating breast cancer by laser-assisted microdissection and quantitative real-time PCR.Pathobiology, 2000,68(45): 173-179.
3. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2~(-△△CT) method. Methods 2001; 25(4): 402-8.
4. Ringrose L, Paro R. Epigenetic regulation of cellular memory by the Polycomb and Trithorax group proteins. Annu Rev Genet, 2004, 38: 413-443.
5. Wang H, Wang L, Erdjument-Bromage H, et al. Tempst P, Jones RS, Zhang Y. Role of histone H2A ubiquitination in Polycomb silencing. Nature, 2004, 431(7010): 873-878.
6. Dahmane N, Sanehez P, Gitton Y, et al. The Sonic Hedgehog-Glipathway regulates dorsal brain growth and tunmorigenesis [J]. Development, 2001,128(24): 5201-5212.
7. Giacinti C, Giordano A. RB and cell cycle progression. Oncogene, 2006; 25(38):5220-5227.
1. Bergqvist D, Lindholm C, Nelzen O, et al. Chronic leg ulcers: the impact of venous disease [J], J Vasc Surg, 1999, 29(8):752-755.
2. Kondo Y, Shen L, Issa J P. Critical role of histone methylation in tumor suppressor gene silencing in colorectal cancer. Mol Cell Biol, 2003, 23(1): 206-215.
3．钟克力，张丽，戴勇等，采用染色质免疫沉淀联合芯片技术分析胃癌全基因组蛋白H3K27三甲基化水平，临床肿瘤杂志，2009．14(1)：15-19。
4. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2~(-△△CT) method. Methods 2001; 25(4): 402-8.
5. Reik W, Dean W. DNA methylation and mammalian epigenetics, Electrophoresis, 2001,22(9): 2838-2843.
6．王志伟，陈玉英，易静等。乳腺癌P16基因甲基化及其临床病理学特征的关系。中华实验外科杂志，2002，19(2)：81。
7．戴观荣，景洪标，邓鉴文。胃癌中脆性组氨酸三联体基因的甲基化和表达及其意义。中华实验外科杂志，2006，23(11)：594-596。
8. Wu J, Wang SH, Potter D, et al. Diverse histone modifications on histone 3 lysine 9 and their relation to DNA methylation in specifying gene silencing. BMC Genomics, 2007,8(3):131.
9. Irvine RA, Lin IG, Hsieh CL. DNA methylation has a local effect on transcription and histone acetylation. Mol Cell Biol, 2002, 22(7):6689-6696.
10. Okitsu CY, Hsieh CL. DNA methylation dictates histone H3K4 methylation. Mol Cell Biol, 2007, 27(8): 2746- 2757.
11．李宇，司聚同，王永潮等。RB、P53、c-myc和Ki-ras基因异常改变在大肠癌发生、 发展中的意义。中华实验外科杂志，1997，14(5)：152。
12. Tamaru H, Selker EU. A histone H3 methyltransferase controls DNA methylation in Neurospora crassa[J]. Nature, 2001, 414(6861):277-83.
13．宋跃，赵青，林海江等。RB基因在结直肠癌中的重排与扩增。白求恩医科大学学报，1998．24(4)：344-346。
14. Kim SK, Hebrok M. Intercellular signals regulating pancreas development and function. Genes Dev 2001; 15:111-127.
1. Wolffe AP, Matzke MA. Epigenetics: regulation through repression [J]. Science,1999,286 (5439): 481-486.
2. Feinberg AP, Vogelstein B. Hypomethylation distinguishes genes of some human cancers from their normal counterparts [J]. Nature, 1983,301 (5895): 89-92.
3． 腾玥，戴冬秋。胃癌表遗传学的研究进展[J]。世界华人消化杂志，2005，13(19)：2289-2293。
4. Sakuma K, Chong JM, Sudo M, et al. High-density methylation of pl4ARF and pl6INK4A in Epstein-Barrvirus-associated gastric car-cinoma. Int JCancer, 2004,112(2): 273-278.
5. Lee YY, Kang SH, Seo JY, et al. Alterations of pl6INK4A and pl5INK4B genes in gastric carcinomas. Cancer, 1997, 80(10): 1889-1896.
6. KanyamaY, HiroshiK, NakayamaH, et al. Detection ofP16 promoter hypermethylation in serum of gastric cancer patients. Cancer Sci, 2003, 94(5): 418-420.
7. EbertMP, Yu J, Hoffmann J, et al. Loss of beta-catenin expression in metastatic gastric cancer. J Clin Oncol. 2003, 21(9): 1708-1714.
8. Byun DS, LeeMG, ChaeKS, et al. Frequent epigenetic inactivation of RASSF1A by aberrant promoter hypermethylation in human gastric adenocarcinoma. Cancer Res,2001, 61 (19): 7034-7038.
9. Fujimoto J, Yasui W, Tahara H, et al. DNA hypermethylation at the pS2 promoter region is associatedwith early stage of stomach carcino-genesis. Cancer Lett, 2000,149(1-2): 125-134.
10. Cunningham JM, Christensen ER, Tester DJ, et al. Hypermethylation of the hMLHl promoter in colon cancerwith microsatellite instabili-ty. Cancer Res, 1998, 58 (15):3455-3460.
11. Rhee I, JairKW, Yen RW, et al. CpG methylation ismaintained in human cancer cells lacking DNMT1. Nature, 2000, 404 (6781): 1003-1007-
12. Xie S, Wang Z, Okano M, et al. Cloning expression and chromosome locations of the human DNMT3 gene family. Gene, 1999, 236(1): 87-95.
13. Toyota M, Ahuja N, OheToyota M, etal. CpG island methylator phe-notype in colorectal cancer. Proc Natl Acad Sci USA, 1999, 96(15): 8681-8686.
14. Mukesh V, Sudhir S. Epigenetics in cancer: implications for early detection and prevention [J]. Lancet Oncol, 2002, 3(7):755-763.
15. Kenneth P, Nephewa, Tim Hui-Ming H. Epigenetic gene silencing in cancer initiation and progression [J]. Cancer Lett, 2003, 190(7): 125-133.
16. Yasui W, Oue N, Ono S, et al. Histone acetylation and gastro-intestinal carcinogenesis [J]. Ann N Y Acad Sci, 2003, 983(3): 220-231.
17. Kim J H, Choi Y K. Downregulation of gelsolin and retinoic acid receptor expression in gastric cancer tissues through histone deacetylase 1[J]. J Gastroenterol Hepatol,2004, 19(2):218-224.
18. Ono S, Oue N, Kuniyasu H. Acetylated histone H4 is reduced in human gastric adenomas and carcinomas [J]. J Exp Clin Cancer Res, 2002, 21(3): 377-382.
19. Zhang Y, Reinberg D. Transcription regulation by histone meth-ylation: interplay between different covalent modifications of the core histone tails [J]. Genes Dev, 2001,15(18): 2343-2360
20. Wysocka J, Swigut T. WDR5 associates with histone H3 meth-ylated at K4 and is essential for H3K4 methylation and verte-brate development [J]. Cell, 2005, 121(6):859-872.
21. Hake S B, Xiao A, Aills C D. Linking the epigenetic language of covalent histone modifications to cancer [J]. British J Cancer, 2004, 90(4): 761-769.
22. Kondo Y, Shen L, Issa JP. Critical role of histone methylation in tumor suppressor gene silencing in colorectal cancer [J]. Mol Cell Biol, 2003, 23(1): 206-215.
23. Cameron E E, Bachman K E, Myohanen S, et al. Synergy of demethylation and histone deacetylase inhibition in there: expression of genes silenced in cancer [J]. Nat Genet,1999, 21(1): 103-107.
24. Tamaru H, Selker EU.A histone H3 methyltransferase controls DNA methylation in Neurospora crassa [J]. Nature, 2001, 414(6861): 277-83.
25. Jenuwein T, Allis CD.Translating the histone code [J]. Science, 2001, 293(5532):1074-80.
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