摘要
前言
2007年,在世界范围内,胃癌仍然是排在第四位的常见恶性肿瘤,估计有100万的新发病例,其中将近70%发生在发展中国家。通常,在亚洲、部分南美国家和一些非洲国家,胃癌的发病率是最高的。在男人和女人的疾病死亡原因中,胃癌分别排在第二位和第四位。2007年,全球范围内大约有80万人死于胃癌。我国胃癌平均年死亡率约为25.2/10万人口。
目前,胃癌的主要治疗措施是手术、化疗和放疗,通常是两种或多种方法相结合。但是,除非早期发现,胃癌难以治愈。在美国,胃癌的5年生存率为24%,如果早期发现,将会提高到61%。采用两种或几种标志物联合检测,有利于提高胃癌的早期诊断率,增进疗效、判断预后和检出复发。目前,CEA和CAl9-9是胃癌的两个主要的肿瘤标志物。发现新的肿瘤标记物是肿瘤的研究方向之一。
CD97分子是表皮生长因子二类七次跨膜受体(EGF-TM7)家族成员之一,拥有独特的杂交结构,N-末端含有3个表皮生长因子(EGF)样功能区,通过粘蛋白样茎区与七次跨膜受体(TM7)偶联。在正常组织中,除小神经胶质细胞以外,CD97只能在巨噬细胞和树突状细胞,以及一些T、B淋巴细胞中丰富表达。如今,CD97被发现在一些上皮性癌组织中丰富表达,如甲状腺、胃肠、胰腺导管和口腔鳞状细胞癌。不同的证据表明,CD97在肿瘤的去分化、迁移、浸润和转移中发挥重要作用。然而,CD97对胃癌细胞生物学特性的影响及其内在机制仍然有待于阐明。
第一章CD97-siRNA转染对人胃癌细胞株MKN45增殖、凋亡、迁移及侵袭能力的影响目的:
设计合成有效的CD97-siRNA,体外转染胃癌细胞株,观察CD97表达变化及其对体内、外胃癌细胞增殖、凋亡、迁移及侵袭能力的影响。
方法:
1.采用人胃癌细胞株MKN45,针对CD97基因设计siRNA,采用化学法合成,筛选出最有效的CD97-siRNA。
2.CD97-siRNA转染胃癌细胞48小时后,用RT-PCR检测CD97基因在mRNA水平表达的变化,72小时后用Western blot和免疫细胞化学方法检测CD97基因在蛋白水平表达的变化,并用MTT法和流式细胞术检测胃癌细胞增殖、凋亡的变化,用细胞迁移和侵袭试验检测胃癌细胞迁移和侵袭能力的变化。
3.15只裸鼠随机分为3组,每组5只,分别接种转染siRNA-CD97细胞(抑制组),接种转染阴性对照siRNA-CD97细胞(阴性对照组),接种未转染细胞(正常对照组)。接种4周后,观察各组成瘤情况,比较成瘤率。
结果:
1.CD97-siRNA转染48小时后,RT-PCR方法检测结果表明CD97在mRNA水平表达下降70.12%;72小时后,Western blot方法检测结果表明CD97在蛋白水平的表达下降66.35%。
2.CD97-siRNA转染72小时后,MTT法检测结果显示,与未转染组相比,细胞增殖率下降52.45%;流式细胞术检测结果显示,G1期细胞数量在未转染组、阴性对照组和CD97-siRNA转染组分别是14.48±3.45%、15.33±2.13%和39.79±3.05%,S期细胞数量分别是64.17±3.36%、60.35±5.21%和28.56±1.33%,出现G1→S期阻滞;同时,细胞凋亡率在未转染组、阴性对照组和CD97-siRNA转染组分别是7.24±0.35%、6.88±0.58和28.18±0.39%。
3.CD97-siRNA转染72小时后,迁移和侵袭实验结果显示CD97-siRNA转染组迁移和侵袭细胞相对于未转染组分别下降了62.54±11.32%和67.13±14.03%。
4.正常对照组和阴性对照组所有裸鼠均有瘤体形成,平均瘤重分别为5.255±0.571克和5.218±0.547克。CD97-siRNA转染组瘤体平均瘤重2.125±0.232克,与正常对照组相比有显著性差异(p<0.05),抑瘤率为58.1%。
结论:
1.CD97蛋白的表达强弱可以作为临床上判断胃癌细胞增殖、转移和侵袭能力的标志物之一,该分子可能成为抑制胃癌复发和转移的重要靶点;
2.RNAi技术可以用于抑制CD97基因的表达;
3.CD97-siRNA转染有望用于胃癌的基因治疗。
第二章CD97-siRNA转染对胃癌细胞株MKN45生物学特性影响的分子机制的初步研究
目的:
研究CD97-siRNA转染诱导MKN45细胞G1期阻滞、细胞凋亡和抑制细胞体外迁移、侵袭能力的分子机制。
方法:
1.Western blot法检测CD97-siRNA转染后对胃癌细胞内细胞周期调节蛋白CyclinD1、CDK4和p27表达的影响。
2.Western blot法检测CD97-siRNA转染后对胃癌细胞内凋亡调节因子Caspase家族表达的影响。
3.Western blot法检测CD97-siRNA转染后对胃癌细胞内凋亡因子Bid、Bad、Bcl-2和Bcl-X_L表达的影响。
4.Western blot法检测CD97-siRNA转染后对胃癌细胞MMP-7和TIMP-1表达的影响。
5.Western blot法检测CD97-siRNA转染后对胃癌细胞PI3K/AKT表达的影响。
结果:
1.Western blot法检测结果表明:与未转染组和阴性对照组相比,CD97-siRNA转染72h后胃癌细胞Cyclin D1、CDK4的表达明显下调,而p27的表达明显上调。
2.Westernblot法检测结果表明:与未转染组和阴性对照组相比,CD97-siRNA转染72h后胃癌细胞procaspase-9、-3的表达水平明显下降,cleaved-caspase-3的表达明显上升,而procaspase-8的表达无明显变化。CD97-siRNA是通过内源性途径诱导凋亡的。
3.Western blot法检测结果表明:与未转染组和阴性对照组相比,CD97-siRNA转染72h后胃癌细胞Bcl-2的表达显著性下调,Bad的表达明显上调,Bcl-xl和Bid的表达无明显变化。
4.Western blot法检测结果表明:与未转染组和阴性对照组相比,CD97-siRNA转染72h后胃癌细胞TIMP-1的表达明显上调,而MMP-7的表达明显下调。
5.Western blot法检测结果表明:与未转染组和阴性对照组相比,CD97-siRNA转染72 h后胃癌细胞PI3K和AKT的磷酸化水平降低,这表明CD97分子可能是通过PI3K/AKT信号通路发挥作用。
结论:
1.CD97受体可能是通过PI3K/AKT信号通路发挥作用的。
2.CD97-siRNA转染下调CD97分子的表达,PI3K及AKT的活性降低,进而上调p27、Bad、TIMP-1蛋白的表达,下调cyclin D1、CDK4、Bcl-2、MMP-7的表达,通过内源性凋亡途径活化caspase-3,从而抑制胃癌细胞的增殖、促进细胞凋亡并抑制肿瘤细胞的迁移、侵袭。
INTRODUCTION
Stomach cancer remained the fourth most common malignancy in the world in 2007,with one million new cases.Nearly 70%of new cases occured in developing countries.Generally,stomach cancer rates are about twice as high in men as in women. About 800,000 people worldwide died from stomach cancer in 2007.It ranks first among all causes of death from cancer in China,with an annual mortality rate of approximately 25.2 per 100.000.
Cancer of the stomach is difficult to cure unless it is found in an early stage.The main treatments for stomach cancer are surgery,chemotherapy,and radiation therapy. Often the best approach uses two or more of these treatment methods.
Detection of two or more than two tumor markers together will increase the rate of early diagnosis of stomach cancer.At present,CEA and CA19-9 are the two main tumor markers of stomach cancer.So,discovery of novel tumor marker is one of the research trend of tumor.
Despite the recently reduced mortality rates due to both earlier detection and improved therapy,gastric cancer is still the second leading cause of cancer-related death worldwide.The development of new treatments is therefore crucial for improving the survival rates of gastric cancer patients.In the last few years,dedifferentiation markers and apoptotic factors have attracted considerable attention as possible targets for the diagnosis and therapy of cancer.
CD97 is a member of a subfamily of classⅡG-protein-coupled receptors referred to as epidermal growth factor seven-transmembrane(EGF-TM7).CD97 protein possesses a unique hybrid structure that consists of varying numbers of N-terminal EGF-like domains coupled to a seven-span TM7 domain by a mucin-like spacer.In normal tissues,abundant expression of CD97 is only detected in macrophages and dendritic cells except for microglial cells,and in some T and B cells.To date,CD97 has been found abundant expression in several epithelial cancers,such as thyroid, gastrointestinal,pancreatic ductal and oral squamous cell carcinoma.Various evidence shows that CD97 plays an important role in tumor dedifferentiation,migration, invasiveness and metastasis.However,the role of CD97 on gastric carcinoma cell growth and the underlying mechanism remain to be elucidated.
Part 1 Effects of CD97-siRNA transfection on biological characteristics in gastric carcinoma cell line MKN45
Objective:
To investigate the effect of CD97 gene silencing by small interfering RNA on proliferation,apoptosis,migration and invasion of gastric carcinoma cell line MKN45 in vivo and in vitro.
Methods:
1.Gastric carcinoma cell line MKN45 was adopted,the pairs of CD97-siRNA were designed according to the CD97 gene sequence and synthesized chemically.
2.Forty-eight hours after CD97-siRNA transfection,the expression of CD97 at mRNA level was detected with RT-PCR.72 hrs after transfection,the expression of CD97 at protein level was detected with Western blot and Immunocytochemistry assays,MTT assay and FCM were used to evaluate the change of proliferation and apoptosis,and Transwell assay was used to evaluate the change of ability of migration and invasion in gastric carcinoma cells.
3.15 nude mice were divided into 3 groups averagely.Group normal control: subcutaneous implant of gastric carcinoma cells with normal gastric carcinoma cells. Group negative control:subcutaneous implant of gastric carcinoma cells with nonsilencing RNA.Group inhibition:subcutaneous implant of gastric carcinoma cells with CD97-siRNA transfection.4 weeks after implantation,the morphology, histopathology and tumor inhibition rate were evaluated on every group.
Results:
1.Forty-eight hours after CD97-siRNA transfection,RT-PCR revealed that CD97 expression decreased 70.12%at mRNA level,and 72 h after CD97-siRNA transfection, Western blot revealed that CD97 expression decreased 66.35%at protein level.
2.Seventy-two hours after CD97-siRNA transfection,MTT revealed that the rate of cell proliferation decreased 52.45%compared to the normal control group,and FCM revealed that the percentage of G1 stage cells of normal control,negative control and transfection group were 14.48±3.45%,15.33±2.13%and 39.79±3.05%respectively;the percentage of S stage cells were 64.17±3.36%,60.35±5.21%and 28.56±1.33% respectively.These data showed that CD97-siRNA restrained the conversion of G1→S stage.FCM revealed that the percentage of apoptotic cells of normal control,negative control and transfection group were 7.24±0.35%,6.88±0.58 and 28.18±0.39% respectively.
3.Seventy-two hours after CD97-siRNA transfection,Transwell assay revealed that the number of migration and invasion cells decreased 62.54±11.32%and 67.13±14.03% respectively.
4.Tumor formation can be found in every nude mice.Tumor tissue mean weights of the normal control and negative control group were 5.255±0.571g and 5.218±0.547g respectively.However,tumor tissue weight of CD97-siRNA transfection group was 2.125±0.232g.Tumor growth was inhibited significantly(p<0.05) and the inhibition rate was 58.1%.
Conclusion:
1.The status of CD97 expression can be used as a marker to evaluate the ability of proliferation,migration and invasion of gastric cancer,and this molecule can be used as an important target to suppress the recurrence and metastasis of gastric cancer.
2.RNAi technique can be used to suppress the expression of CD97 gene.
3.CD97-siRNA may be used as gene therapy means for gastric cancer.
Part 2 A pilot study of the underlying molecular mechanism of CD97-siRNA transfection-induced biological characteristics in gastric carcinoma cell line MKN45
Objective:
To study the underlying molecular mechanism of CD97-siRNA-induced cell cycle arrest, apoptosis,and suppression of migration and invasion of gastric carcinoma cell line MKN45 in vitro.
Methods:
1.Western blot assay was used to determine the change of level of protein of Cyclin D1、CDK4 and p27 after CD97-siRNA transfection.
2.Western blot assay was used to determine the change of level of protein of caspase family after CD97-siRNA transfection.
3.Western blot assay was used to determine the change of level of protein of Bid、Bad、Bcl-2 and Bcl-X_L after CD97-siRNA transfection.
4.Western blot assay was used to determine the change of level of protein of MMP-7 and TIMP-1 after CD97-siRNA transfection.
5.Western blot assay was used to determine the effect of CD97-siRNA transfection on PI3K/AKT signal pathway.
Results:
1.Western blot assay revealed that CD97-siRNA transfection down-regulated the level of expression of Cyclin D1 and CDK4,and up-regulated the level of expression of p27 in MKN45 cells significantly.
2.Western blot assay revealed that CD97-siRNA transfection down-regulated the level of expression of procaspase-9,-3,and up-regulated the level of expression of cleaved-caspase-3 significantly,however,had no significant influence on the level of expression of procaspase-8 in MKN45 cells.These data showed that CD97-siRNA triggers apoptosis of gastric cancer cells largely through the intrinsic pathway.
3.Western blot assay revealed that CD97-siRNA transfection down-regulated the level of expression of Bcl-2 and up-regulated the level of expression of Bad significantly, however,had no significant influence on the level of expression of Bcl-x1 and Bid in MKN45 cells.
4.Western blot assay revealed that CD97-siRNA transfection up-regulated the level of expression of TIMP-1,down-regulated the level of expression of MMP-7 in MKN45 cells significantly.
5.Western blot assay revealed that CD97-siRNA transfection decreased the amount of phosphorylated PI3K and AKT in MKN45 cells significantly.These data showed that CD97 enhance the cancer growth probably through PI3K/AKT signal transduction pathway.
Conclusion:
1.The CD97 receptor enhance the cancer growth probably through PI3K/AKT signal transduction pathway.
2.CD97-siRNA transfection may down-regulate the level of expression of CD97 gene, inactivate the PI3K and AKT,up-regulate the level of expression of p27, cleaved-caspase-3,Bad and TIMP-1,and down-regulate the level of expression of cyclin D1,CDK4,procaspase-9,-3,Bcl-2 and MMP-7,furtherly result in cell cycle arrest,apoptosis,and suppression of migration and invasion of gastric carcinoma cells.
引文
1. Garcia M JA,Ward EM,Center MM,Hao Y,Siegel RL,Thun MJ.:Global Cancer Facts & Figures 2007.Atlanta,GA:American Cancer Society,2007 2007.
2. 樊丹：恶性肿瘤是城市居民首位死因．中医药管理杂志 2008,16(5):1.
3. Sun XD,Mu R,Zhou YS,Dai XD,Zhang SW,Huangfu XM,Sun J,Li LD,Lu FZ,Qiao YL:[Analysis of mortality rate of stomach cancer and its trend in twenty years in China].Zhonghua Zhong Liu Za Zhi 2004,26(1):4-9.
4. Yasui W,Sentani K,Motoshita J,Nakayama H:Molecular pathobiology of gastric cancer.Scand J Surg 2006,95(4):225-231.
5. Iwasaki Y,Arai K,Katayanagi S,Takahashi K,Yamaguchi T,Matsumoto H,Miyamoto H:[Biomarkers for neoplasmas in digestive organs].Gan To Kagaku Ryoho 2004,31(7):1015-1020.
6. Meknight AJ GS:The EGF-TM7 family:unusual structures at the leucocyte surface.J Leukoc Biol 1998,63(3):10.
7. Hamann J EW,Hamann D,Kerstens HM,Poddighe PJ,Hoovers JM,,Hartmann E SM,van Lier RA:Expression cloning and chromosomal mapping of the leukocyte activation antigen CD97,a new seven-span transmembrane molecule of the secretion receptor superfamily with an unusual extracellular domain..J Immunol 1995,155(4):9.
8. Baud V,Chissoe SL,Viegas-Pequignot E,Diriong S,N'Guyen VC,Roe BA,Lipinski M:EMR1,an unusual member in the family of hormone receptors with seven transmembrane segments.Genomics 1995,26(2):334-344.
9. Lin HH,Stacey M,Hamann J,Gordon S,McKnight AJ:Human EMR2,a novel EGF-TM7 molecule on chromosome 19p13.1,is closely related to CD97.Genomics 2000,67(2):188-200.
10. Stacey M,Lin HH,Hilyard KL,Gordon S,McKnight AJ:Human epidermal growth factor(EGF)module-containing mucin-like hormone receptor 3 is a new member of the EGF-TM7 family that recognizes a ligand on human macrophages and activated neutrophils.J Biol Chem 2001,276(22):18863-18870.
11. Caminschi I,Lucas KM,O'Keeffe MA,Hochrein H,Laabi Y,Kontgen F,Lew AM,Shortman K,Wright MD:Molecular cloning of F4/80-like-receptor,a seven-span membrane protein expressed differentially by dendritic cell and monocyte-macrophage subpopulations.J Immunol 2001,167(7):3570-3576.
12. Nechiporuk T,Urness LD,Keating MT:ETL,a novel seven-transmembrane receptor that is developmentally regulated in the heart.ETL is a member of the secretin family and belongs to the epidermal growth factor-seven-transmembrane subfamily.J Biol Chem 2001,276(6):4150-4157.
13. Kwakkenbos MJ,Kop EN,Stacey M,Matmati M,Gordon S,Lin HH,Hamann J:The EGF-TM7 family:a postgenomic view,lmmunogenetics 2004,55(10):655-666.
14. Hamann J,Hartmann E,van Lier RA:Structure of the human CD97 gene:exon shuffling has generated a new type of seven-span transmembrane molecule related to the secretin receptor superfamily.Genomics 1996,32(1):144-147.
15. Jaspars LH,Vos W,Aust G,Van Lier RA,Hamann J:Tissue distribution of the human CD97 EGF-TM7 receptor.Tissue Antigens 2001,57(4):325-331.
16. Gray JX,Haino M,Roth MJ,Maguire JE,Jensen PN,Yarme A,Stetler-Stevenson MA,Siebenlist U,Kelly K:CD97 is a processed,seven-transmembrane,heterodimeric receptor associated with inflammation.J Immunol 1996,157(12):5438-5447.
17. Aust G,Eichler W,Laue S,Lehmann I,Heldin NE,Lotz O,Scherbaum WA,Dralle H,Hoang-Vu C:CD97:a dedifferentiation marker in human thyroid carcinomas.Cancer Res 1997,57(9):1798-1806.
18. Hoang-Vu C,Bull K,Schwarz I,Krause G,Schmutzler C,Aust G,Kohrle J,Dralle H:Regulation of CD97 protein in thyroid carcinoma.J Clin Endocrinol Metab 1999,84(3):1104-1109.
19. Steinert M,Wobus M,Boltze C,Schutz A,Wahlbuhl M,Hamann J,Aust G:Expression and regulation of CD97 in colorectal carcinoma cell lines and tumor tissues.Am J Pathol 2002,161(5):1657-1667.
20. Aust G,Steinert M,Schutz A,Boltze C,Wahlbuhl M,Hamann J,Wobus M:CD97,but not its closely related EGF-TM7 family member EMR2,is expressed on gastric,pancreatic,and esophageal carcinomas.Am J Clin Pathol 2002,118(5):699-707.
21. Mustafa T,Klonisch T,Hombach-Klonisch S,Kehlen A,Schmutzler C,Koehrle J,Gimm O,Dralle H,Hoang-Vu C:Expression of CD97 and CD55 in human medullary thyroid carcinomas.Int J Oncol 2004,24(2):285-294.
22. Mustafa T,Eckert A,Klonisch T,Kehlen A,Maurer P,Klintschar M,Erhuma M,Zschoyan R,Gimm O,Dralle H et al:Expression of the epidermal growth factor seven-transmembrane member CD97 correlates with grading and staging in human oral squamous cell carcinomas.Cancer Epidemiol Biomarkers Prev 2005,14(1):108-119.
23. Gravalos C,Jimeno A:HER2 in gastric cancer:a new prognostic factor and a novel therapeutic target.Ann Oncol 2008,19(9):1523-1529.
24. Blume-Jensen P,Hunter T:Oncogenic kinase signalling.Nature 2001,411(6835):355-365.
25. Muller H,Lukas J,Schneider A,Warthoe P,Bartek J,Eilers M,Strauss M:Cyclin D1 expression is regulated by the retinoblastoma protein.Proc Natl Acad Sci USA 1994,91(8):2945-2949.
26. Baldin V,Lukas J,Marcote MJ,Pagano M,Draetta G:Cyclin Dl is a nuclear protein required for cell cycle progression in G1.Genes Dev 1993,7(5):812-821.
27. Gravalos C,Jimeno A:HER2 in gastric cancer:a new prognostic factor and a novel therapeutic target.Ann Oncol 2008,19(9):1523-1529.
28. Won KA,Xiong Y,Beach D,Gilman MZ:Growth-regulated expression of D-type cyclin genes in human diploid fibroblasts.Proc Natl Acad Sci USA 1992,89(20):9910-9914.
29. Lukas J,Pagano M,Staskova Z,Draetta G,Bartek J:Cyclin Dl protein oscillates and is essential for cell cycle progression in human tumour cell lines.Oncogene 1994,9(3):707-718.
30. Huang Z:Bcl-2 family proteins as targets for anticancer drug design.Oncogene 2000,19(56):6627-6631.
31. Yonemura Y,Endou Y,Fujita H,Fushida S,Bandou E,Taniguchi K,Miwa K,Sugiyama K,Sasaki T:Role of MMP-7 in the formation of peritoneal dissemination in gastric cancer.Gastric Cancer 2000,3(2):63-70.
32. Fire A,Xu S,Montgomery MK,Kostas SA,Driver SE,Mello CC:Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans.Nature 1998,391(6669):806-811.
33. Dalmay T:MicroRNAs and cancer.J Intern Med 2008,263(4):366-375.
34. Lipardi C,Wei Q,Paterson BM:RNAi as random degradative PCR:siRNA primers convert mRNA into dsRNAs that are degraded to generate new siRNAs.Cell 2001,107(3):297-307.
35. Pierce KL,Premont RT,Lefkowitz RJ:Seven-transmembrane receptors.Nat Rev Mol Cell Biol 2002,3(9):639-650.
36. Brzostowski JA,Kimmel AR:Signaling at zero G:G-protein-independent functions for 7-TM receptors.Trends Biochem Sci 2001,26(5):291-297.
37. Segre GV,Goldring SR:Receptors for secretin,calcitonin,parathyroid hormone(PTH)/PTH-related peptide,vasoactive intestinal peptide,glucagonlike peptide 1,growth hormone-releasing hormone,and glucagon belong to a newly discovered G-protein-linked receptor family.Trends Endocrinol Metab 1993,4(10):309-314.
38. Zhang X,Jin B,Huang C:The PI3K/Akt pathway and its downstream transcriptional factors as targets for chemoprevention.Curr Cancer Drug Targets 2007,7(4):305-316.
39. Eichler W,Hamann J,Aust G:Expression characteristics of the human CD97 antigen.Tissue Antigens 1997,50(5):429-438.
40. Rajagopal R,Chao MV:A role for Fyn in Trk receptor transactivation by G-protein-coupled receptor signaling.Mol Cell Neurosci 2006,33(1):36-46.
41. Aust G SM,Schutz A,Boltze C,Wahlbuhl M,Hamann J,Wobus M.:CD97,but not its closely related EGF-TM7 family member EMR2,is expressed on gastric,pancreatic,and esophageal carcinomas.Am J Clin Pathol 2002,118(5):9.
42. Steinert M,Wobus M,Boltze C,Schutz A,Wahlbuhl M,Hamann J,Aust G:Expression and regulation of CD97 in colorectal carcinoma cell lines and tumor tissues.Am J Pathol 2002,161(5):1657-1667.
43. Aust G,Eichler W,Laue S,Lehmann I,Heldin NE,Lotz O,Scherbaum WA,Dralle H,Hoang-Vu C:CD97:a dedifferentiation marker in human thyroid carcinomas.Cancer Res 1997,57(9):1798-1806.
44.Liu Y,Chen L,Peng S-Y,Chen Z-X,Hoang-Vu C:Role of CD97(stalk) and CD55 as molecular markers for prognosis and therapy of gastric carcinoma patients.J Zhejiang Univ Sci B 2005,6(9):913-918.
45.柳咏,陈力,彭淑牖:胃癌组织中CD97stalk和CD55的表达及其临床病理学意义.实用肿瘤杂志2005,20(4):5.
46.Liu Y,Chen L,Peng S,Chen Z,Gimm O,Finke R,Hoang-Vu C:The expression of CD97EGF and its ligand CD55 on marginal epithelium is related to higher stage and depth of tumor invasion of gastric carcinomas.Oncol Rep 2005,14(6):1413-1420.
47.Lin CH,Fu ZM,Liu YL,Yang JL,Xu JF,Chen QS,Chen HM:Investigation of SGC-7901 cell line established from human gastric carcinoma cells.Chin Med J(Engl) 1984,97(11):831-834.
48.Boring CC,Squires TS,Tong T,Montgomery S:Cancer statistics,1994.CA Cancer J Clin 1994,44(1):7-26.
49.McKnight AJ,Gordon S:The EGF-TM7 family:unusual structures at the leukocyte surface.J Leukoc Biol 1998,63(3):271-280.
50.Loberg RD,Wojno KJ,Day LL,Pienta KJ:Analysis of membrane-bound complement regulatory proteins in prostate cancer.Urology 2005,66(6):1321-1326.
51.Galle J,Sittig D,Hanisch I,Wobus M,Wandel E,Loeffler M,Aust G:Individual cell-based models of tumor-environment interactions:Multiple effects of CD97 on tumor invasion.Am J Pathol 2006,169(5):1802-1811.
52.Kop EN,Adriaansen J,Smeets TJ,Vervoordeldonk MJ,van Lier RA,Hamann J,Tak PP:CD97neutralisation increases resistance to collagen-induced arthritis in mice.Arthritis Res Ther 2006,8(5):R155.
53.Muller H,Lukas J,Schneider A,Warthoe P,Bartek J,Eilers M,Strauss M:Cyclin D1expression is regulated by the retinoblastoma protein.Proc Natl Acad Sci U S A 1994,91(8):2945-2949.
54.Baldin V,Lukas J,Marcote MJ,Pagano M,Draetta G:Cyclin Dl is a nuclear protein required for cell cycle progression in G1.Genes Dev 1993,7(5):812-821.
55.Motokura T,Keyomarsi K,Kronenberg HM,Arnold A:Cloning and characterization of human cyclin D3,a cDNA closely related in sequence to the PRAD1/cyclin D1proto-oncogene.JBiol Chem 1992,267(28):20412-20415.
56.Dillon RL,White DE,Muller WJ:The phosphatidyl inositol 3-kinase signaling network:implications for human breast cancer.Oncogene 2007,26(9):1338-1345.
57.Pierce KL,Premont RT,Lefkowitz RJ:Seven-transmembrane receptors.Nat Rev Mol Cell Biol 2002,3(9):639-650.
58. Brzostowski JA,Kimmel AR:Signaling at zero G:G-protein-independent functions for 7-TM receptors.Trends Biochem Sci 2001,26(5):291-297.
59. Segre GV,Goldring SR:Receptors for secretin,calcitonin,parathyroid hormone(PTH)/PTH-related peptide,vasoactive intestinal peptide,glucagonlike peptide 1,growth hormone-releasing hormone,and glucagon belong to a newly discovered G-protein-linked receptor family.Trends Endocrinol Metab 1993,4(10):309-314.
60. Polyak K,Kato JY,Solomon MJ,Sherr CJ,Massague J,Roberts JM,Koff A:p27Kipl,a cyclin-Cdk inhibitor,links transforming growth factor-beta and contact inhibition to cell cycle arrest.Genes Dev 1994,8(1):9-22.
61. Pietenpol JA,Bohlander SK,Sato Y,Papadopoulos N,Liu B,Friedman C,Trask BJ,Roberts JM,Kinzler KW,Rowley JD et at Assignment of the human p27Kipl gene to 12p13 and its analysis in leukemias.Cancer Res 1995,55(6):1206-1210.
62. Kravtsov VG,Shakhmurov MG,Sukmanov OV,Zairat'iants OV,Shirin NI:[Expression of cycline-dependent kinase p27 in the low differentiated gastric adenocarcinoma].Arkh Patol 2006,68(5):14-16.
63. Zheng JY,Wang WZ,Li KZ,Guan WX,Yan W:Effect of p27(KIPl)on cell cycle and apoptosis in gastric cancer cells.World J Gastroenterol 2005,11(45):7072-7077.
64. Fulda S,Debatin KM:Extrinsic versus intrinsic apoptosis pathways in anticancer chemotherapy.Oncogene 2006,25(34):4798-4811.
65. Walczak H,Krammer PH:The CD95(APO-1/Fas)and the TRAIL(APO-2L)apoptosis systems.Exp Cell Res 2000,256(1):58-66.
66. Saelens X,Festjens N,Vande Walle L,van Gurp M,van Loo G,Vandenabeele P:Toxic proteins released from mitochondria in cell death.Oncogene 2004,23(16):2861-2874.
67. McDonnell TJ,Beham A,Sarkiss M,Andersen MM,Lo P:Importance of the Bcl-2 family in cell death regulation.Experientia 1996,52(10-11):1008-1017.
68. Reed JC:Proapoptotic multidomain Bcl-2/Bax-family proteins:mechanisms,physiological roles,and therapeutic opportunities.Cell Death Differ 2006,13(8):1378-1386.
69. Yoshikawa T,Tsuburaya A,Kobayashi O,Sairenji M,Motohashi H,Yanoma S,Noguchi Y:Intratumoral concentrations of tissue inhibitor of matrix metalloproteinase 1 in patients with gastric carcinoma a new biomartker for invasion and its impact on survival.Cancer 2001,91(9):1739-1744.
70. Yoon SO,Shin S,Lee HJ,Chun HK,Chung AS:Isoginkgetin inhibits tumor cell invasion by regulating phosphatidylinositol 3-kinase/Akt-dependent matrix metalloproteinase-9 expression.Mol Cancer Ther 2006,5(11):2666-2675.
71. Cheng JC,Chou CH,Kuo ML,Hsieh CY:Radiation-enhanced hepatocellular carcinoma cell invasion with MMP-9 expression through PI3K/Akt/NF-kappaB signal transduction pathway.Oncogene 2006,25(53):7009-7018.
72. Bae IH,Park MJ,Yoon SH,Kang SW,Lee SS,Choi KM,Um HD:Bcl-w promotes gastric cancer cell invasion by inducing matrix metalloproteinase-2 expression via phosphoinositide 3-kinase,Akt,and Sp1.Cancer Res 2006,66(10):4991-4995.
73. Aust G SM,Schutz A,Boltze C,Wahlbuhl M,Hamann J,Wobus M.:CD97,but not its closely related EGF-TM7 family member EMR2,is expressed on gastric,pancreatic,and esophageal carcinomas.Am J Clin Pathol 2002,118(5):9.
74. Meetoo D:Chronic diseases:the silent global epidemic.BrJNurs 2008,17(21):1320-1325.
75. Brody T,Cravchik A:Drosophila melanogaster G protein-coupled receptors.J Cell Biol 2000,150(2):F83-88.
76. Lander ES,Linton LM,Birren B,Nusbaum C,Zody MC,Baldwin J,Devon K,Dewar K,Doyle M,FitzHugh W et al:Initial sequencing and analysis of the human genome.Nature 2001,409(6822):860-921.
77. Venter JC,Adams MD,Myers EW,Li PW,Mural RJ,Sutton GG,Smith HO,Yandell M,Evans CA,Holt RA et al:The sequence of the human genome.Science 2001,291(5507):1304-1351.
78. Waterston RH,Lindblad-Toh K,Birney E,Rogers J,Abril JF,Agarwal P,Agarwala R,Ainscough R,Alexandersson M,An P et al:Initial sequencing and comparative analysis of the mouse genome.Nature 2002,420(6915):520-562.
79. Genome sequence of the nematode C.elegans:a platform for investigating biology.Science 1998,282(5396):2012-2018.
80. Bockaert J,Pin JP:Molecular tinkering of G protein-coupled receptors:an evolutionary success.EMBO J 1999,18(7):1723-1729.
81. Harmar AJ:Family-B G-protein-coupled receptors.GenomeBiol200\,2(12):REVIEWS3013.
82. Stacey M,Lin HH,Gordon S,McKnight AJ:LNB-TM7,a group of seven-transmembrane proteins related to family-B G-protein-coupled receptors.Trends Biochem Sci 2000,25(6):284-289.
83. Fredriksson R,Gloriam DE,Hoglund PJ,Lagerstrom MC,Schioth HB:There exist at least 30 human G-protein-coupled receptors with long Ser/Thr-rich N-termini.Biochem Biophys Res Commun 2003,301(3):725-734.
84. Hayflick JS:A family of heptahelical receptors with adhesion-like domains:a marriage between two super families.J Recept Signal Transduct Res 2000,20(2-3):119-131.
85. Hamann J,Eichler W,Hamann D,Kerstens HM,Poddighe PJ,Hoovers JM,Hartmann E,Strauss M,van Lier RA:Expression cloning and chromosomal mapping of the leukocyte activation antigen CD97,a new seven-span transmembrane molecule of the secretion receptor superfamily with an unusual extracellular domain.J Immunol 1995,155(4):1942-1950.
86. Hamann J,Kwakkenbos MJ,de Jong EC,Heus H,Olsen AS,van Lier RA:Inactivation of the EGF-TM7 receptor EMR4 after the Pan-Homo divergence.Eur J Immunol 2003, 33(5):1365-1371.
87. Bork P,Downing AK,Kieffer B,Campbell ID:Structure and distribution of modules in extracellular proteins.Q Rev Biophys 1996,29(2):119-167.
88. Hamann J,Vogel B,van Schijndel GM,van Lier RA:The seven-span transmembrane receptor CD97 has a cellular ligand(CD55,DAF).J Exp Med 1996,184(3):1185-1189.
89. Lublin DM,Atkinson JP:Decay-accelerating factor:biochemistry,molecular biology,and function.Annu Rev Immunol 1989,7:35-58.
90. Mikesch JH,Buerger H,Simon R,Brandt B:Decay-accelerating factor(CD55):a versatile acting molecule in human malignancies.Biochim Biophys Acta 2006,1766(1):42-52.
91. Hamann J,Stortelers C,Kiss-Toth E,Vogel B,Eichler W,van Lier RA:Characterization of the CD55(DAF)-binding site on the seven-span transmembrane receptor CD97.Eur J Immunol 1998,28(5):1701-1707.
92. Hamann J,van Zeventer C,Bijl A,Molenaar C,Tesselaar K,van Lier RA:Molecular cloning and characterization of mouse CD97.Int Immunol 2000,12(4):439-448.
93. Lin HH,Stacey M,Saxby C,Knott V,Chaudhry Y,Evans D,Gordon S,McKnight AJ,Handford P,Lea S:Molecular analysis of the epidermal growth factor-like short consensus repeat domain-mediated protein-protein interactions:dissection of the CD97-CD55 complex.J Biol Chem 2001,276(26):24160-24169.
94. Leemans JC,te Velde AA,Florquin S,Bennink RJ,de Bruin K,van Lier RA,van der Poll T,Hamann J:The epidermal growth factor-seven transmembrane(EGF-TM7)receptor CD97 is required for neutrophil migration and host defense.J Immunol 2004,172(2):1125-1131.
95. Stacey M,Chang GW,Davies JQ,Kwakkenbos MJ,Sanderson RD,Hamann J,Gordon S,Lin HH:The epidermal growth factor-like domains of the human EMR2 receptor mediate cell attachment through chondroitin sulfate glycosaminoglycans.Blood2003,102(8):2916-2924.
96. Kwakkenbos MJ,Chang GW,Lin HH,Pouwels W,de Jong EC,van Lier RA,Gordon S,Hamann J:The human EGF-TM7 family member EMR2 is a heterodimeric receptor expressed on myeloid cells.J Leukoc Biol 2002,71(5):854-862.
97. Trowbridge JM,Gallo RL:Dermatan sulfate:new functions from an old glycosaminoglycan.Glycobiology 2002,12(9):117R-125R.
98. Eichler W,Aust G,Hamann D:Characterization of an early activation-dependent antigen on lymphocytes defined by the monoclonal antibody BL-Ac(F2).Scand J Immunol 1994,39(1):111-115.
99. Schlossman SF BL,Gilks W,Harlan JM,Kishimoto T,Morimoto C,Ritz J,Shaw S,Silverstein R,Springer TA,Tedder TF,Todd RF Leukocyte typing V:white cell differentiation antigens.Oxford:Oxford University Press;1995.
100. Visser L,de Vos AF,Hamann J,Melief MJ,van Meurs M,van Lier RA,Laman JD,Hintzen RQ:Expression of the EGF-TM7 receptor CD97 and its ligand CD55(DAF)in multiple sclerosis.J Neuroimmunol 2002,132(1-2):156-163.
101. Hamann J,Wishaupt JO,van Lier RA,Smeets TJ,Breedveld FC,Tak PP:Expression of the activation antigen CD97 and its ligand CD55 in rheumatoid synovial tissue.Arthritis Rheum 1999,42(4):650-658.
102. Firestein GS,Zvaifler NJ:How important are T cells in chronic rheumatoid synovitis?:II.T cell-independent mechanisms from beginning to end.Arthritis Rheum 2002,46(2):298-308.
103. Gasz B,Lenard L,Benko L,Borsiczky B,Szanto Z,Lantos J,Szabados S,Alotti N,Papp L,Roth E:Expression of CD97 and adhesion molecules on circulating leukocytes in patients undergoing coronary artery bypass surgery.Eur Surg Res 2005,37(5):281-289.
104. Veninga H,Becker S,Hoek RM,Wobus M,Wandel E,van der Kaa J,van der Valk M,de Vos AF,Haase H,Owens B et at Analysis of CD97 expression and manipulation:antibody treatment but not gene targeting curtails granulocyte migration.J Immunol 2008,181(9):6574-6583.
105. Zhou H,Xu M,Huang Q,Gates AT,Zhang XD,Castle JC,Stec E,Ferrer M,Strulovici B,Hazuda DJ et at Genome-scale RNAi screen for host factors required for HIV replication.Cell Host Microbe 2008,4(5):495-504.
106. Aust G,Steinert M,Schutz A,Boltze C,Wahlbuhl M,Hamann J,Wobus M:CD97,but not its closely related EGF-TM7 family member EMR2,is expressed on gastric,pancreatic,and esophageal carcinomas.Am J Clin Pathol 2002,118(5):699-707.
107. Wang T,Ward Y,Tian L,Lake R,Guedez L,Stetler-Stevenson WQ Kelly K:CD97,an adhesion receptor on inflammatory cells,stimulates angiogenesis through binding integrin counterreceptors on endothelial cells.Blood 2005,105(7):2836-2844.
108. Wobus M,Huber O,Hamann J,Aust G:CD97 overexpression in tumor cells at the invasion front in colorectal cancer(CC)is independently regulated of the canonical Wnt pathway.Mol Carcinog 2006,45(11):881-886.
109. Zhang X,Jin B,Huang C:The PI3K/Akt pathway and its downstream transcriptional factors as targets for chemoprevention.Curr Cancer Drug Targets 2007,7(4):305-316.
110. Rajagopal R,Chao MV:A role for Fyn in Trk receptor transactivation by G-protein-coupled receptor signaling.Mol Cell Neurosci 2006,33(1):36-46.
111. Tahara E:Molecular mechanism of stomach carcinogenesis.J Cancer Res Clin Oncol 1993,119(5):265-272.
112. Yasui W,Oue N,Kuniyasu H,Ito R,Tahara E,Yokozaki H:Molecular diagnosis of gastric cancer:present and future.Gastric Cancer2001,4(3):113-121.
113. Vainio H HE:Handbook of experimental pharmacology.Mechanisms in carcinogenesis and cancer research.Berlin Heidelberg New York Singapore Tokyo:Springer-Verlag 2003.
114. Yokozaki H,Yasui W,Tahara E:Genetic and epigenetic changes in stomach cancer.Int Rev Cytol 2001,204:49-95.
115. Yasui W,Oue N,Ono S,Mitani Y,Ito R,Nakayama H:Histone acetylation and gastrointestinal carcinogenesis.Ann N Y Acad Sci 2003,983:220-231.
116. E T:Molecular pathology of gastroenterological cancer.Berlin Heidelberg Now York Singapore Tokyo:Springer-Verlag;1997.
117. Anderson NL,Anderson NG:The human plasma proteome:history,character,and diagnostic prospects.Mol Cell Proteomics 2002,1(11):845-867.
118. Srivastava S,Verma M,Henson DE:Biomarkers for early detection of colon cancer.Clin Cancer Res 2001,7(5):1118-1126.
119. Stoss O,Henkel,T:Biomedical marker molecules for cancer-current status and perpectives.Drug Discovery Today:Targets 2004,3:10.
120. Wagner PD,Maruvada P,Srivastava S:Molecular diagnostics:a new frontier in cancer prevention.Expert Rev Mol Diagn 2004,4(4):503-511.
121. Bangma CH,Grobbee DE,Schroder FH:Volume adjustment for intermediate prostate-specific antigen values in a screening population.Eur J Cancer 1995,31A(1):12-14.
122. Gann PH,Hennekens CH,Stampfer MJ:A prospective evaluation of plasma prostate-specific antigen for detection of prostatic cancer.JAMA 1995,273(4):289-294.
123. Gillatt D,Reynard JM:What is the 'normal range' for prostate-specific antigen? Use of a receiver operating characteristic curve to evaluate a serum marker.Br J Urol 1995,75(3):341-346.
124. Lepor H,Owens RS,Rogenes V,Kuhn E:Detection of prostate cancer in males with prostatism.Prostate 1994,25(3):132-140.
125. Biomarkers and surrogate endpoints:preferred definitions and conceptual framework.Clin Pharmacol Ther 2001,69(3):89-95.
126. Pollak MN,Foulkes WD:Challenges to cancer control by screening.Nat Rev Cancer 2003,3(4):297-303.
127. Neugut AI,Jacobson JS,Rella VA:Prevalence and incidence of colorectal adenomas and cancer in asymptomatic persons.Gastrointest Endosc Clin N Am 1997,7(3):387-399.
128. Manne U,Srivastava RG,Srivastava S:Recent advances in biomarkers for cancer diagnosis and treatment.Drug Discov Today 2005,10(14):965-976.
129. Wagner PD,Verma M,Srivastava S:Challenges for biomarkers in cancer detection.Ann N Y Acad Sci 2004,1022:9-16.
130. Mihmanli M,Dilege E,Demir U,Coskun H,Eroglu T,Uysalol MD:The use of tumor markers as predictors of prognosis in gastric cancer.Hepatogastroenterology 2004,51(59):1544-1547.
131. Yamamoto M,Baba H,Toh Y,Okamura T,Maehara Y:Peritoneal lavage CEA/CA125 is a prognostic factor for gastric cancer patients.J Cancer Res Clin Oncol 2007,133(7):471-476.
132. Goral V,Yesilbagdan H,Kaplan A,Sit D:Evaluation of CA 72-4 as a new tumor marker in patients with gastric cancer.Hepatogastroenterology 2007,54(76):1272-1275.
133. Werner M,Becker KF,Keller G,Hofler H:Gastric adenocarcinoma:pathomorphology and molecular pathology.J Cancer Res Clin Oncol 2001,127(4):207-216.
134. Takahashi Y,Cleary KR,Mai M,Kitadai Y,Bucana CD,Ellis LM:Significance of vessel count and vascular endothelial growth factor and its receptor(KDR)in intestinal-type gastric cancer.Clin Cancer Res 1996,2(10):1679-1684.
135. Kitadai Y,Haruma K,Sumii K,Yamamoto S,Ue T,Yokozaki H,Yasui W,Ohmoto Y,Kajiyama G,Fidler IJ et ah Expression of interleukin-8 correlates with vascularity in human gastric carcinomas.Am J Pathol 1998,152(1):93-100.
136. Takahashi Y,Bucana CD,Akagi Y,Liu W,Cleary KR,Mai M,Ellis LM:Significance of platelet-derived endothelial cell growth factor in the angiogenesis of human gastric cancer.Clin Cancer Res 1998,4(2):429-434.
137. Kido S,Kitadai Y,Hattori N,Haruma K,Kido T,Ohta M,Tanaka S,Yoshihara M,Sumii K,Ohmoto Y et at Interleukin 8 and vascular endothelial growth factor — prognostic factors in human gastric carcinomas? Eur J Cancer 2001,37(12):1482-1487.
138. Yasui W,Yokozaki H,Shimamoto F,Tahara H,Tahara E:Molecular-pathological diagnosis of gastrointestinal tissues and its contribution to cancer histopathology.Pathol Int 1999,49(9):763-774.
139. Xiangming C,Natsugoe S,Takao S,Hokita S,Tanabe G,Baba M,Kuroshima K,Aikou T:The cooperative role of p27 with cyclin E in the prognosis of advanced gastric carcinoma.Cancer 2000,89(6):1214-1219.
140. Tsujie M,Yamamoto H,Tomita N,Sugita Y,Ohue M,Sakita I,Tamaki Y,Sekimoto M,Doki Y,Inoue M et al:Expression of tumor suppressor gene p16(INK4)products in primary gastric cancer.Oncology 2000,58(2):126-136.
141. Feakins RM,Nickols CD,Bidd H,Walton SJ:Abnormal expression of pRb,p16,and cyclin D1 in gastric adenocarcinoma and its lymph node metastases:relationship with pathological features and survival.Hum Pathol 2003,34(12):1276-1282.
142. Fondevila C,Metges JP,Fuster J,Grau JJ,Palacin A,Castells A,Volant A,Pera M:p53 and VEGF expression are independent predictors of tumour recurrence and survival following curative resection of gastric cancer.Br J Cancer 2004,90(1):206-215.
143. Pinto-de-Sousa J,Silva F,David L,Leitao D,Seixas M,Pimenta A,Cardoso-de-Oliveira M:Clinicopathological significance and survival influence of p53 protein expression in gastric carcinoma.Histopathology 2004,44(4):323-331.
144. Shimada Y,Yamasaki S,Hashimoto Y,Ito T,Kawamura J,Soma T,Ino Y,Nakanishi Y,Sakamoto M,Hirohashi S et al:Clinical significance of dysadherin expression in gastric cancer patients.Clin Cancer Res 2004,10(8):2818-2823.
145. Chan AO,Lam SK,Chu KM,Lam CM,Kwok E,Leung SY,Yuen ST,Law SY,Hui WM,Lai KC et al:Soluble E-cadherin is a valid prognostic marker in gastric carcinoma.Gut 2001, 48(6):808-811.
146. Yamamichi K,Uehara Y,Kitamura N,Nakane Y,Hioki K:Increased expression of CD44v6 mRNA significantly correlates with distant metastasis and poor prognosis in gastric cancer.Int J Cancer 1998,79(3):256-262.
147. Saito H,Tsujitani S,Katano K,Ikeguchi M,Maeta M,Kaibara N:Serum concentration of CD44 variant 6 and its relation to prognosis in patients with gastric carcinoma.Cancer 1998,83(6):1094-1101.
148. Mayer B,Jauch KW,Gunthert U,Figdor CG,Schildberg FW,Funke I,Johnson JP:De-novo expression of CD44 and survival in gastric cancer.Lancet 1993,342(8878):1019-1022.
149. Yamashita K,Azumano I,Mai M,Okada Y:Expression and tissue localization of matrix metalloproteinase 7(matrilysin)in human gastric carcinomas.Implications for vessel invasion and metastasis.Int J Cancer 1998,79(2):187-194.
150. Inoue T,Yashiro M,Nishimura S,Maeda K,Sawada T,Ogawa Y,Sowa M,Chung KH:Matrix metalloproteinase-1 expression is a prognostic factor for patients with advanced gastric cancer.Int J Mol Med 1999,4(1):73-77.
151. Gao ZL,Zhang C,Du GY,Lu ZJ:Clinical significance of changes in tumor markers,extracellular matrix,MMP-9 and VEGF in patients with gastric carcinoma.Hepatogastroenterology 2007,54(77):1591-1595.
152. Mori M,Mimori K,Shiraishi T,Fujie T,Baba K,Kusumoto H,Haraguchi M,Ueo H,Akiyoshi T:Analysis of MTl-MMP and MMP2 expression in human gastric cancers.Int J Cancer 1997,74(3):316-321.
153. Mimori K,Mori M,Shiraishi T,Fujie T,Baba K,Haraguchi M,Abe R,Ueo H,Akiyoshi T:Clinical significance of tissue inhibitor of metalloproteinase expression in gastric carcinoma.Br J Cancer 1997,76(4):531-536.
154. Kim NW,Piatyszek MA,Prowse KR,Harley CB,West MD,Ho PL,Coviello GM,Wright WE,Weinrich SL,Shay JW:Specific association of human telomerase activity with immortal cells and cancer.Science 1994,266(5193):2011-2015.
155. Yasui W,Tahara E,Tahara H,Fujimoto J,Naka K,Nakayama J,Ishikawa F,Ide T:Immunohistochemical detection of human telomerase reverse transcriptase in normal mucosa and precancerous lesions of the stomach.Jpn J Cancer Res 1999,90(6):589-595.
156. Kondo T,Oue N,Yoshida K,Mitani Y,Naka K,Nakayama H,Yasui W:Expression of POT1 is associated with tumor stage and telomere length in gastric carcinoma.Cancer Res 2004,64(2):523-529.
157. Horii A,Han HJ,Shimada M,Yanagisawa A,Kato Y,Ohta H,Yasui W,Tahara E,Nakamura Y:Frequent replication errors at microsatellite loci in tumors of patients with multiple primary cancers.Cancer Res 1994,54(13):3373-3375.
158. Keller G,Rotter M,Vogelsang H,Bischoff P,Becker KF,Mueller J,Brauch H,Siewert JR, Hofler H:Microsatellite instability in adenocarcinomas of the upper gastrointestinal tract.Relation to clinicopathological data and family history.Am J Pathol 1995,147(3):593-600.
159. Takahashi H,Endo T,Yamashita K,Arimura Y,Yamamoto H,Sasaki S,Itoh F,Hirata K,Imamura A,Kondo M et al:Mucin phenotype and microsatellite instability in early multiple gastric cancers.Int J Cancer 2002,100(4):419-424.
160. dos Santos NR,Seruca R,Constancia M,Seixas M,Sobrinho-Simoes M:Microsatellite instability at multiple loci in gastric carcinoma:clinicopathologic implications and prognosis.Gastroenterology 1996,110(1):38-44.
161. Iacopetta BJ,Soong R,House AK,Hamelin R:Gastric carcinomas with microsatellite instability:clinical features and mutations to the TGF-beta type II receptor,IGFII receptor,and BAX genes.J Pathol 1999,187(4):428-432.
162. Wirtz HC,Muller W,Noguchi T,Scheven M,Ruschoff J,Hommel G,Gabbert HE:Prognostic value and clinicopathological profile of microsatellite instability in gastric cancer.Clin Cancer Res 1998,4(7):1749-1754.
163. Choi SW,Choi JR,Chung YJ,Kim KM,Rhyu MG:Prognostic implications of microsatellite genotypes in gastric carcinoma.Int J Cancer 2000,89(4):378-383.
164. Mitani Y,Oue N,Hamai Y,Aung PP,Matsumura S,Nakayama H,Kamata N,Yasui W:Histone H3 acetylation is associated with reduced p21(WAFl/CIPl)expression by gastric carcinoma.J Pathol 2005,205(1):65-73.
165. Suzuki T,Yokozaki H,Kuniyasu H,Hayashi K,Naka K,Ono S,Ishikawa T,Tahara E,Yasui W:Effect of trichostatin A on cell growth and expression of cell cycle-and apoptosis-related molecules in human gastric and oral carcinoma cell lines.Int J Cancer 2000,88(6):992-997.
166. Oue N,Motoshita J,Yokozaki H,Hayashi K,Tahara E,Taniyama K,Matsusaki K,Yasui W:Distinct promoter hypermethylation of p16INK4a,CDH1,and RAR-beta in intestinal,diffuse-adherent,and diffuse-scattered type gastric carcinomas.J Pathol 2002,198(1):55-59.
167. Oshimo Y,Oue N,Mitani Y,Nakayama H,Kitadai Y,Yoshida K,Ito Y,Chayama K,Yasui W:Frequent loss of RUNX3 expression by promoter hypermethylation in gastric carcinoma.Pathobiology 2004,71(3):137-143.
168. Cai JC,Liu D,Zhang HP,Zhong S,Xia NS:[Promoter methylation of several tumor suppressor genes in human gastric adenocarcinoma].Zhonghua Yi Xue Za Zhi 2007,87(14):978-981.
169. Oue N,Matsumura S,Nakayama H,Kitadai Y,Taniyama K,Matsusaki K,Yasui W:Reduced expression of the TSP1 gene and its association with promoter hypermethylation in gastric carcinoma.Oncology 2003,64(4):423-429.
170. Hamai Y,Oue N,Mitani Y,Nakayama H,Ito R,Matsusaki K,Yoshida K,Toge T,Yasui W:DNA hypermethylation and histone hypoacetylation of the HLTF gene are associated with reduced expression in gastric carcinoma.Cancer Sci 2003,94(8):692-698.
171. Oshimo Y,Oue N,Mitani Y,Nakayama H,Kitadai Y,Yoshida K,Chayama K,Yasui W:Frequent epigenetic inactivation of RIZl by promoter hypermethylation in human gastric carcinoma.Int J Cancer 2004,110(2):212-218.
172. Oshimo Y,Kuraoka K,Nakayama H,Kitadai Y,Yoshida K,Chayama K,Yasui W:Epigenetic inactivation of SOCS-1 by CpG island hypermethylation in human gastric carcinoma.Int J Cancer 2004,112(6):1003-1009.
173. Toyota M,Ahuja N,Suzuki H,Itoh F,Ohe-Toyota M,Imai K,Baylin SB,Issa JP:Aberrant methylation in gastric cancer associated with the CpG island methylator phenotype.Cancer Res 1999,59(21):5438-5442.
174. Yasui W,Oue N,Aung PP,Matsumura S,Shutoh M,Nakayama H:Molecular-pathological prognostic factors of gastric cancer:a review.Gastric Cancer 2005,8(2):86-94.
175. Gonzalez CA,Sala N,Capella G:Genetic susceptibility and gastric cancer risk.Int J Cancer 2002,100(3):249-260.
176. Schneider J,Schulze G:Comparison of tumor M2-pyruvate kinase(tumor M2-PK),carcinoembryonic antigen(CEA),carbohydrate antigens CA 19-9 and CA 72-4 in the diagnosis of gastrointestinal cancer.Anticancer Res 2003,23(6D):5089-5093.