膜联蛋白A5在散发性大肠癌中的表达及其与大肠癌分期及预后的相关性研究
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摘要
近年来,我国大肠癌的发病率呈上升趋势,且早期症状少,待出现症状后才明确者多属中晚期,故其是一种恶性程度较高的消化道肿瘤,死亡率位居西方发达国家恶性肿瘤死亡率的第2、3位,在发展中国家发病率也呈逐年升高趋势。据调查显示,目前大肠癌在中国大部分地区已经成为发病率上升最快的恶性肿瘤之一。2002年以后上海市大肠癌发病率已经超越胃癌跃居第2位,仅次于肺癌。当前,我们治疗大肠癌的方法相对成熟,主要是以手术切除为主,辅助以化疗、放疗或免疫治疗,以及中药治疗等。但大肠癌术后5年内的复发率、死亡率仍然保存在一个较高的水平。早期发现并采取积极有效的治疗措施成为广大医务工作者的共同目标,因此寻找作为大肠癌早期诊断的新型肿瘤标志物及新的大肠癌治疗靶标差异蛋白,成为目前大肠癌基础及临床研究的热点。本研究我们关注的是膜联蛋白A家族的一个重要成员--膜联蛋白A5。
     膜联蛋白是一种以钙离子依赖性方式与酸性磷脂结合的蛋白质,在大多数生物的细胞中,膜联蛋白是一种高丰度蛋白,占整个细胞蛋白质含量的0.5%~2%,其中膜联蛋白A家族成员广泛分布于高等脊椎动物中。大量的研究显示,膜联蛋白在胞吞、胞吐作用、细胞间以及细胞与细胞外基质间的相互作用、离子通道形成、磷脂酶A2活性抑制等多种生理过程中起作用,但是二十多年来的研究并没有能够就其生理功能得到一个明确结论。长期以来人们发现膜联蛋白A家族成员与不同来源的良性、恶性肿瘤的发病机制相关,研究显示在多种恶性肿瘤组织中A家族某些成员的表达水平都发生了明显的上调,然而另外一些资料显示在某些恶性肿瘤生成、分化以及转移过程中膜联蛋白的下调可能起着重要的作用。遗憾的是迄今为止膜联蛋白A家族成员与肿瘤的发生、发展的确切关系及具体作用机制仍然不明确。
     相关研究显示膜联蛋白A家族某些成员的表达在不同特征的大肠癌组织中发生了变化,提示它们可能在大肠癌的发生、发展过程中扮演者重要角色,然而并没有强有力的证据证实它们在大肠癌发病过程中所起的作用,甚至于不同的研究者得到了互相矛盾的结果。因此我们认为有必要借助于一些较先进的技术手段,对膜联蛋白A家族成员在大肠癌中的表达情况及其作用机制进行系统、深入的探讨。
     首先,我们由正常人胎盘组织中克隆得到了膜联蛋白A5的基因编码区全长序列,并通过亚克隆技术将其装载入真核表达载体pcDNA3.1(+)。
     我们收集了51例病例完整、背景清楚的散发性大肠癌患者组织样本作为研究对象,同时获得了51例非恶性肠道疾病患者的组织样本作为对照。通过免疫组织化学实验,我们首先对肿瘤样本中p53、Ki-67这两个肿瘤临床诊断上较为常用的标志物进行了分析,并分别根据p53的突变情况和Ki-67增殖指数对51例肿瘤组织样本进行了归类。随后我们选择了目前功能较为清楚的膜联蛋白A5作为切入点,借助荧光实时定量PCR和免疫组织化学等实验手段,分别从mRNA水平和蛋白质水平上对膜联蛋白A5在组织样本中的表达情况进行了检测。研究结果显示:膜联蛋白A5的表达水平在散发性大肠癌组织中发生了上调;同时A5在肿瘤组织中的表达变化与p53的突变和Ki-67增殖指数分别呈正相关。这一结果提示我们膜联蛋白A5与大肠癌的恶化、增殖等特征具有相关性,可能在其中起着重要的作用,并且膜联蛋白A5能够作为一种肿瘤标志物应用于大肠癌的临床诊断,因此具有临床应用价值。
     在此基础上,我们扩大样本量,选择了176例病例完整、背景清楚的散发性大肠癌患者组织样本作为研究对象。手术切除标本后,分别配对取其大肠癌变部位组织以及正常组织。然后通过实时定量PCR和免疫组化染色的方法鉴定每个标本的膜联蛋白A5的mRNA和蛋白表达情况。实时定量PCR结果显示肿瘤中的膜联蛋白A5的mRNA水平上调。免疫组化结果显示膜联蛋白A5在40.5%的肿瘤之中高表达。同时进行严格详细的术后随访。通过统计数据判断膜联蛋白A5的表达与病理分期和预后之间的关系。我们的研究结果显示,膜联蛋白A5的表达情况可以作为一个有价值的分子标记物,用来预测患者外科手术切除肿瘤后的预后。无论是进行单变量分析还是多变量分析,我们的研究数据都显示,膜联蛋白A5的表达情况和患者5年生存率的降低以及肿瘤复发率的提高都有很强的相关性。多变量分析显示,膜联蛋白A5的表达情况可以作为一个独立的预测因子用来预测预后。因此,不考虑肿瘤的病理分期而仅用肿瘤组织中的膜联蛋白A5表达水平也可以预测患者的术后预后。另外,膜联蛋白A5表达水平也可作为术前预测治疗效果的一个预测因子。我们的研究提示,膜联蛋白A5在大肠癌的生长、进展以及转移等过程中可能扮演了一个相当重要的角色。
     在组织水平研究的基础之上,我们选择了两种遗传背景相同转移潜能不同的大肠癌细胞系(高转移潜能:SW620、低转移潜能:SW480)。首先,通过Transwell小室实验对两种细胞系的侵袭、迁移能力进行检测,以验证之前对它们转移潜能的分析。随后通过荧光实时定量PCR和Western blot从mRNA水平和蛋白水平对膜联蛋白A家族重要成员A1、A2和A5的表达情况进行了检测。结果显示不同的A家族成员在具备不同转移潜能的大肠癌细胞系中的表达情况互有差异(A1在高转移潜能的细胞中较高、A2在低转移潜能的细胞系中较高、A5的表达与细胞转移潜能不具明显相关性),这一结果提示我们A家族膜联蛋白与肿瘤细胞的转移、凋亡等多个方面具有相关性。我们设想膜联蛋白A家族不同成员在各生理过程中的重要性并不相同,可能存在功能上的互补性。
     综上所述,本研究对膜联蛋白A5在不同特征的散发性大肠癌中的表达情况进行了系统的检测,对其在大肠癌发生、发展中的作用机制进行了初步分析,并且通过分析膜联蛋白A5的表达与肿瘤分期及临床病理特征之间的相关性,探讨了膜联蛋白A5作为标志物预测患者预后的潜在可能性。并为更加深入地探讨膜联蛋白A家族与大肠癌的发生、发展关系以及膜联蛋白的生理功能研究提供较好的理论依据。
Carcinoma of the colon and rectum is one of the most common alimentary malignancies in developed nations, and cancer mortality among the first places 2,3. Even in developing countries, morbidity is with a increasing trend. Now, CRC has become one of malignancies with the fastest rising incidence in most parts of China. In Shanghai, 2002, colorectal cancer incidence has gone beyond the first two ranks, second only to lung cancer. Today , operation is the most important treatment of CRC, supported by chemotherapy, radiotherapy or immunotherapy, as well as traditional Chinese medicine treatment. However, colorectal cancer after 5 years the relapse rate, mortality still remains at a higher level. To take positive measures for early detection and effective treatment has become the common goal for medical workers. So looking for new tumor markers for early diagnosis of colorectal cancer and new protein for therapeutic targets is a hot spots for basic and clinical research of CRC.
     This study we are concerned is one important member of annexin A family ---Annexin A5. The annexins are characterized as a family of proteins capable of binding to acidic phospholipids in a Ca2+-dependent manner and they appear to be widely distributed throughout nature. Up to this day, there are total of twelve annexins have been found in higher vertebrates and form the A family of the annexins. In most biological contexts studied, the annexins are highly abundant proteins, reaching the level as high as 0.5% to 2% of the total cellular proteins and all these proteins present a homologous core structure and a highly variable N-terminus. Based on their Ca2+-dependent interaction with phospholipids and membranes, many physiological functions have been described for the annexins including endocytosis, exocytosis, cell-cell or cell-matrix interaction, formation of ion channels, inhibition of phospholipase A2 and so on. Unfortunately, there has not been a definitive description of any physiological role played by these annexins in more than 20 years of study. Although any exact in physiological function has not been described for the annexins, there are great deals of evidence to suggest that they are involved in the pathogenesis of many different cancers. In many cancers, there is sharp up-regulation of annexins in both mRNA and protein levels. On the other hand, there is data indicating that down-regulation of annexins may play a significant role in tumorigenesis and metastasis of other types of cancer. However, the precise role played by the annexins in the pathogenesis of tumor is still unknown.
     Related studies have shown that expression of some members of annexin A family has changed in the colorectal cancer tissues with different characteristics. That means these members have been playing important roles in the occurrence and the development process of colorectal cancer. Nevertheless, no strong evidences had shown they had affected incidence of colorectal cancer, even different researchers had conflicting results.Therefore, in our opinion, it is necessary to analyze the expression of A family members in colorectal cancer and their mechanism in tumorigenesis.
     Above all, we succeeded in cloning full length sequence of human annexin A5 coding regions of from human placenta by RT-PCR. Then we constructed and identified recombinant eukaryotic expression vectors of annexin A5.
     51 tumor samples with complete clinical data were selected. At the same time 51 normal tissues (obtained from patients with benign colorectal diseases) were chosen as normal control. We firstly detected p53 and Ki-67 in tumor tissues by immunoassaying and split all tumor samples into two groups separately according to the detection of p53 or Ki-67. Then the mRNA and protein levels of annexin A5 in each sample were analyzed by real-time quantitative PCR and immunohistochemistry. We found that there is an up-regulation of annexin A5 in colorectal cancer. We also found the up-regulation has a positive correlation with the mutation of p53 or Labeling Index (Ki-67) in tumor tissues. Besides, the mutation of p53 in tumor tissues has a negative correlation with Labeling Index (Ki-67). Our study indicates that annexin A5 might be used as an assistant clinicopathological biomarker in cancer diagnosis and the up-regulation of annexin A5 in colorectal cancer might be related to the malignant proliferation of tumor cells.
     We investigated the expression of ANXA5 in colorectal adenocarcinoma. This study included 176 consecutive patients with sporadic colorectal cancer. Paired colorectal tissue samples and corresponding nonmalignant tissues were obtained by surgical resection. ANXA5 mRNA and protein expression in each tissue were assessed by real-time RT-PCR and immunohistochemical staining. Real-time RT-PCR showed that there is an up-regulation in mRNA level of ANXA5 in tumors . Immunohistochemical study revealed that high ANXA5 expression was present in 40.58% of tumors. Data were statistically correlated with pathological parameters and clinical outcome. In this study, we have shown that expression of ANXA5 can be used as a potential molecular marker to predict patient outcome in patients treated with surgical resection alone. Our data showed that high ANXA5 expression has a strong correlation with decreased five-year survival and increased tumor recurrence both in univariate and multivariate analyses. Multivariate analysis indicated that ANXA5 expression is an independent prognostic factor for poor survival. Thus, tumor levels of ANXA5 regardless of tumor stage can be used to determine patient outcome. In addition, ANXA5 expression level can potentially be used as a prognostic factor before surgery can be done. Our findings also suggest that ANXA5 plays an important role in tumor growth, progression, and metastasis in colorectal adenocarcinoma.
     We selected two human colon adenocarcinoma cell lines and analyzed their metastatic potentials by invasion/migration assay (higher metastatic potential: SW620; lower metastatic potential:SW480). Then we analyzed expression of annexin A1, A2 and A5 mRNA and protein levels in cell lines by real-time PCR and western blot. We found that the order of A1, A2, A5 expression level from high to low was separately SW620 >SW480, SW480 > SW620, and SW480≥SW620. Our results indicated that annexin A1 and A2 may play important roles in tumor metastasis and annexin A5 may not work in metastasis but apoptosis. We also imagine that different annexins may be complementary in function
     We analyzed expression of annexin A5 in spocitic colorectal cancer and its mechnium in tumorigenesis. We determined its prognostic significance by correlating ANXA5 expression with tumor stage and clinicopathologic features.Our work would make the groundwork for further researches on physiological functions of the annexins and their role in tumorigenesis.
引文
[1] Lee DB, Jamgotchian N, Allen SG, et al. Annexin A2 heterotetramer: role in tight junction assembly. Am J Physiol Renal Physiol. 2004 Sep; 287(3): F481-491. Epub 2004 Apr 27.
    [2] Hawkins TE, Merrifield CJ, Moss SE. Calcium signaling and annexins. Cell Biochem Biophys. 2000; 3(3): 275-296.
    [3] Gerke V, Moss SE. Annexins and membrane dynamics. Biochim Biophys Acta. 1997 Jun 27; 1357(2): 129-154.
    [4] Gerke V, Moss SE. Annexins: from structure to function. Physiol Rev. 2002 Apr; 82(2): 331-371.
    [5] Iwasaki A, Suda M, Nakao H, et al. Structure and expression of cDNA for an inhibitor of blood coagulation isolated from human placenta: a new lipocortin-like protein. J Biochem (Tokyo). 1987 Nov; 102(5): 1261-1273.
    [6] Parente L, Solito E. Annexin 1: more than an anti-phospholipase protein. Inflamm Res. 2004 Apr; 53(4): 125-132. Epub 2004 Mar 18.
    [7] Raynal P, Pollard HB. Annexins: the problem of assessing the biological role for a gene family of multifunctional calcium- and phospholipid-binding proteins. Biochim Biophys Acta. 1994 Apr 5; 1197(1): 63-93.
    [8] Kamangar F,Dores GM,Anderson WF.Patterns of cancer incidence,mortality,and prevalence across five continents:defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 2006;24:2137-2150.
    [9] Eilstein D,Hedelin G,Schaffer P.Incidence of colorectal cancer in Bas-Rhin,trend and prediction in 2009.Bull cancer 2000;87:595-599.
    [10] Ji BT,Devesa SS,Chow WH,Jin F,Gao YT.Colorectal cancer incidence trends by subsite in urban shanghai,1992-1994.Cancer Epidemiol Biomarkers Prev 1998;7:661-666.
    [11] Emoto K, Yamada Y, Sawada H, et al. Annexin II overexpression correlates with stromal tenascin-C overexpression: a prognostic marker in colorectal carcinoma. Cancer. 2001 Sep 15; 92(6): 1419-1426.
    [12] Sun M, Liu Y, Gibb W. Distribution of annexin I and II in term human fetal membranes, decidua and placenta. Placenta. 1996 Mar-Apr; 17(2-3): 181-184.
    [13] Hawkins TE, Merrifield CJ, Moss SE. Calcium signaling and annexins. Cell Biochem Biophys. 2000; 33(3): 275-96.
    [14] Morgan RO, Pilar Fernandez M. Distinct annexin subfamilies in plants and protists diverged prior to animal annexins and from a common ancestor. J Mol Evol. 1997 Feb; 44(2): 178-88.
    [15] Morgan RO, Bell DW, Testa JR, et al. Human annexin 31 genetic mapping and origin. Gene. 1999 Feb 4; 227(1): 33-8.
    [16] Liu W, Saint DA. Validation of a quantitative method for real time PCR kinetics. Biochem Biophys Res Commun. 2002 Jun 7; 294(2): 347-353.
    [17] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001 Dec; 25(4): 402-408.
    [18] Luecke H, Chang BT, Mailliard WS, et al. Crystal structure of the annexin XII hexamer and implications for bilayer insertion. Nature. 1995 Nov 30; 378(6556): 512-515.
    [19] Sudo T, Hidaka H. Regulation of calcyclin (S100A6) binding by alternative splicing in the N-terminal regulatory domain of annexin XI isoforms. J Biol Chem. 1998 Mar 13; 273(11): 6351-6357.
    [20] Huber R,Berendes R,Burger A,et al:Crystal structure and its implication on function.In:Moss SE,ed.The Annexins.London:Portland Press,2001:105-124.
    [21] Liemann S,Lewit-Bentley A.Annexins:A novel family of calcium-and membrane-binding proteins in serch of a function.Structure 1995;3:233-237.
    [22] Swairjo MA,Seaton BA.Annexin structure and membrane interactions:A molecular prespective..Anuu Rev Biophys Biomol Struct 1994;23:193-213.
    [23] Weng X, Luecke H, Song IS, et al. Crystal structure of human annexin I at 2.5 A resolution. Protein Sci. 1993 Mar; 2(3): 448-458.
    [24] Rescher U, Zobiack N, Gerke V. Intact Ca(2+)-binding sites are required for targeting of annexin 1 to endosomal membranes in living HeLa cells. J Cell Sci. 2000 Nov; 113 (Pt 22): 3931-3938.
    [25] Eberhard D, Karns LR, Vandenberg SR, et al. Control of the nuclear-cytoplasmic partitioning of annexin II by a nuclear export signal and by pll binding. J Cell Sci. 2001 Sep; 114(Pt 17): 3155-3166.
    [26] Kourie JI, Wood HB. Biophysical and molecular properties of annexin-formed channels. Prog Biophys Mol Biol. 2000; 73(2-4): 91-134.
    [27] Swairjo MA, Concha NO, Kaetzel MA et al. Ca2+-bridging mechanism and phospholipids head group recognition in the membrane-binding protein annexin V. Nat Struct Biol 1995;2:968-974.
    [28] Rand JH. Antiphospholipid antibody-mediated disruption of the annexin-V antithrombotic shield: a thrombogenic mechanism for the antiphospholipid syndrome. J Autoimmun. 2000 Sep; 15(2): 107-111.
    [29] von der Mark K, Mollenhauer J. Annexin V interactions with collagen. Cell Mol Life Sci. 1997 Jun; 53(6): 539-545.
    [30] Schlaepfer DD, Jones J, Haigler HT. Inhibition of protein kinase C by annexin V. Biochemistry 1992; 31:1886-1891.
    [31] Rothhut B, Dubois T, Feliers D et al. Inhibitory effect of annexin V on protein kinase C activity in mesangial cell lysates. Eur J Biochem 1995; 232:865-872.
    [32] Wen Y, Edelman JL, Kang T et al. Lipocortin V may function as a signaling protein for vascular endothelial growth factor receptor-2/Flk-1. Biochem Biophys Res Commun 1999; 258:713-721.
    [33] Barwise JL, Walker JH. Annexins II, IV, V and VI relocate in response to rises in intracellular calcium in human foreskin fibroblasts. J Cell Sci 1996; 109:247-255.
    [34] Tzima E, Trotter PJ, Hastings AD et al. Investigation of the relocation of cytosolic phospholipase A2 and annexin V in activated platelets. Thromb Res 2000; 97:421-429.
    [35] Tzima E, Trotter PJ, Orchard MA et al. Annexin V relocates to the platelet cytoskeleton upon activation and binds to a specific isoform of actin. Eur J Biochem 2000; 267:4720-4730.
    [36] Lee TL, Lin YC, Mochitate K, et al. Stress-relaxation of fibroblasts in collagen matrices triggers ectocytosis of plasma membrane vesicles containing actin, annexins II and VI, and beta 1 integrin receptors. J Cell Sci. 1993 May; 105 (Pt 1): 167-177.
    [37] Olse E,Rasmussen HH,Celis JE.Identification of proteins that are abnormally regulated in differentiated cultured human keratinocutes,Electrophoresis 1995;16:2241-2248.
    [38] Paweletz CP,Ornstein DK,Roth MJ et al.Loss of annexin I correlates with early onset of yumorigenesis in esophageal and prostate carcinoma.Cancer Res 2000;60:6293-6297.
    [39] Genge BR, Cao X, Wu LN et al. Establishment of the primary structure of the major lipid-dependent Ca2+ binding proteins of chicken growth plate cartilage matrix vesicles: identity with anchorin CII (annexin V) and annexin II. J Bone Miner Res 1992; 7:807-819.
    [40] De Meyer S, Gong ZJ, Suwandhi W et al. Organ and species specificity of hepatitis B virus (HBV) infection: A review of literature with aspecial reference to preferential attachment of HBV to human hepatocytes. J Viral Hepat 1997; 4:145-153.
    [41] von der Mark K, Mollenhauer J. Annexin V interaction with collagen. Cell Mol Life Sci 1997; 53:539-545.
    [42] van Heerde WL, Robert-Offerman S, Dumont E et al. Markers of apoptosis in cardiovascular tissues: focus on annexin V. Cardiovasc Res 2000; 45:549-559.
    [43] Meycalfe P, Williamson LM, Reutelingsperger CP et al. Activation during preparation of therapeutic platelets affects deterioration during storage: A comparative flow cytometric study of different production methods. Br J Haematol 1997; 98:86-95.
    [44] Stratton JR, Dewhurst TA,ina S etal. Selective uptake of radiolabeled annexin V on acute porcine left atrial thrombi. Circulation 1995; 92:3113-3121.
    [45] Doring V, Veretout F, Albrecht R, et al. The in vivo role of annexin VII (synexin): characterization of an annexin VII-deficient Dictyostelium mutant indicates an involvement in Ca(2+)-regulated processes. J Cell Sci. 1995 May; 108 (Pt 5): 2065-2076.
    [46] Hawkins TE, Roes J, Rees D, et al. Immunological development and cardiovascular function are normal in annexin VI null mutant mice. Mol Cell Biol. 1999 Dec; 19(12): 8028-8032.
    [47] Srivastava M, Atwater I, Glasman M, et al. Defects in inositol 1,4,5-trisphosphate receptor expression, Ca(2+) signaling, and insulin secretion in the anx7(+/-) knockout mouse. Proc Natl Acad Sci U S A. 1999 Nov 23; 96(24): 13783-137888.
    [48] Herr C, Smyth N, Ullrich S, et al. Loss of annexin A7 leads to alterations in frequency-induced shortening of isolated murine cardiomyocytes. Mol Cell Biol. 2001 Jul; 21(13): 4119-41.
    [1] Gerke V, Moss SE. Annexins: from structure to function. Physiol Rev. 2002 Apr; 82(2): 331-371.
    [2] Rescher U, Gerke V. Annexins--unique membrane binding proteins with diverse functions. J Cell Sci. 2004 Jun 1; 117(Pt 13): 2631-2639.
    [3] Lorusso A, Covino C, Priori G, et al. Annexin2 coating the surface of enlargeosomes is needed for their regulated exocytosis. EMBO J. 2006 Nov 29; 25(23): 5443-5456. Epub 2006 Nov 2.
    [4] Kenis H, van Genderen H, Deckers NM, et al. Annexin A5 inhibits engulfment through internalization of PS-expressing cell membrane patches. Exp Cell Res. 2006 Apr 1; 312(6): 719-726. Epub 2005 Dec 27.
    [5] Turnay J, Olmo N, Lizarbe MA, et al. Changes in the expression of annexin A5 gene during in vitro chondrocyte differentiation: influence of cell attachment. J Cell Biochem. 2001; 84(1): 132-142.
    [6] Gerke V, Creutz CE, Moss SE. Annexins: linking Ca2+ signalling to membrane dynamics. Nat RevMol Cell Biol. 2005 Jun; 6(6): 449-461.
    [7] Buckland AG, Wilton DC. Inhibition of secreted phospholipases A2 by annexin V. Competition for anionic phospholipid interfaces allows an assessment of the relative interfacial affinities of secreted phospholipases A2. Biochim Biophys Acta. 1998 Apr 22; 1391(3): 367-376.
    [8] Gilmore WS, Olwill S, McGlynn H, et al. Annexin A2 expression during cellular differentiation in myeloid cell lines. Biochem Soc Trans. 2004 Dec; 32(Pt 6): 1122-1133.
    [9] Monceau V, Belikova Y, Kratassiouk G, et al. Externalization of endogenous annexin A5 participates in apoptosis of rat cardiomyocytes. Cardiovasc Res. 2004 Dec 1; 64(3): 496-506.
    [10] Hayes MJ, Moss SE. Annexins and disease. Biochem Biophys Res Commun. 2004 Oct 1; 322(4): 1166-1170.
    [11] Watson AJ. An overview of apoptosis and the prevention of colorectal cancer. Crit Rev Oncol Hematol. 2006 Feb; 57(2): 107-21. Epub 2005 Dec 1.
    [12] de la Chapelle A. Genetic predisposition to colorectal cancer. Nat Rev Cancer. 2004 Oct; 4(10): 769-80.
    [13] Alldridge LC, Bryant CE. Annexin 1 regulates cell proliferation by disruption of cell morphology and inhibition of cyclin D1 expression through sustained activation of the ERK1/2 MAPK signal. Exp Cell Res. 2003 Oct 15; 290(1): 93-107.
    [14] Singh P. Role of Annexin-II in GI cancers: Interaction with gastrins/progastrins. Cancer Lett. 2006 Dec 21; [Epub ahead of print]
    [15] Grewal T, Enrich C. Molecular mechanisms involved in Ras inactivation: the annexin A6-p120GAP complex. Bioessays. 2006 Dec; 28(12): 1211-1220.
    [16] Emoto K, Yamada Y, Sawada H, et al. Annexin II overexpression correlates with stromal tenascin-C overexpression: a prognostic marker in colorectal carcinoma. Cancer. 2001 Sep 15; 92(6): 1419-1426.
    [17] Liu W, Saint DA. Validation of a quantitative method for real time PCR kinetics. Biochem Biophys Res Commun. 2002 Jun 7; 294(2): 347-253.
    [18] Kirsch DG, Kastan MB. Tumor-suppressor p53: implications for tumor development and prognosis. J Clin Oncol. 1998 Sep; 16(9): 3158-3168.
    [19] Munro AJ, Lain S, Lane DP. P53 abnormalities and outcomes in colorectal cancer: a systematic review. Br J Cancer. 2005 Feb 14; 92(3): 434-444.
    [20] Endl E, Gerdes J. The Ki-67 protein: fascinating forms and an unknown function. Exp Cell Res. 2000 Jun 15; 257(2): 231-237.
    [21] Papantoniou VJ, Souvatzoglou MA, Valotassiou VJ, et al. Relationship of cell proliferation (Ki-67) to 99mTc-(V)DMSA uptake in breast cancer. Breast Cancer Res. 2004; 6(2): R56-62. Epub 2003 Dec 11.
    [22] Komori K, Ajioka Y, Watanabe H, et al. Proliferation kinetics and apoptosis of serrated adenoma of the colorectum. Pathol Int. 2003 May; 53(5): 277-283.
    [23] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 2001 Dec; 25(4): 402-8.
    [24] Karube A, Shidara Y, Hayasaka K, et al. Suppression of calphobindin I (CPB I) production in carcinoma of uterine cervix and endometrium. Gynecol Oncol. 1995 Sep; 58(3): 295-300.
    [25] Ahmed FE. Molecular markers that predict response to colon cancer therapy. Expert Rev Mol Diagn. 2005 May; 5(3): 353-375.
    [26] Hattori T, Hoffman T, Hirata F. Differentiation of a histiocytic lymphoma cell line by lipomodulin, a phospholipase inhibitory protein. Biochem Biophys Res Commun. 1983 Mar 16 ;111(2): 551-9.
    [27] Emoto K, Yamada Y, Sawada H, Annexin II overexpression correlates with stromal tenascin-C overexpression :a prognostic marker in colorectal poor carcinoma. Cancer 2001 sep 15; 92(6): 1419-1426.
    [28] Emoto K, Sawada H, Yamada Y, Annexin II overexpression is correlated with poor prognosis in human gastric carcinoma. Anticancer Res. 2001 Mar-Apr; 21(2B): 1339-1345.
    [29] Frohlich M, Motte P, Galvin K, et al. Enhanced expression of the protein kinase substrate p36 in human hepatocellular carcinoma. Mol Cell Biol. 1990 Jun; 10(6): 3216-3223.
    [30] Manda R, Kohno T, Matsuno Y, et al. Identification of genes (SPON2 and C20orf2) differentially expressed between cancerous and noncancerous lung cells by mRNA differential display. Genomics. 1999 Oct 1 ;61(1): 5-14.
    [31] Kumble KD, Hirota M, Pour PM, et al. Enhanced levels of annexins in pancreatic carcinoma cells of Syrian hamsters and their intrapancreatic allografts. Cancer Res. 1992 Jan 1; 52(1): 163-7.
    [32] Vishwanatha JK, Chiang Y, Kumble KD, Enhanced expression of annexin II in human pancreatic carcinoma cells and primary pancreatic cancers. Carcinogenesis. 1993 Dec; 14(12): 2575-9.
    [33] Golias CH, Charalabopoulos A, Charalabopoulos K. Cell proliferation and cell cycle control: a mini review. Int J Clin Pract. 2004 Dec; 58(12): 1134-41.
    [34] Li W, Sanki A, Karim RZ, et al. The role of cell cycle regulatory proteins in the pathogenesis of melanoma. Pathology. 2006 Aug; 38(4): 287-301.
    [35] Sun Y. p53 and its downstream proteins as molecular targets of cancer. Mol Carcinog. 2006 Jun; 45(6): 409-15.
    [36] Johannessen AL, Torp SH. The clinical value of Ki-67/MIB-1 labeling index in human astrocytomas. Pathol Oncol Res. 2006; 12(3): 143-147. Epub 2006 Sep 23.
    [37] Haut Donahue TL, Genetos DC, Jacobs CR, Annexin V disruption impairs mechanically inducedcalcium signaling in osteoblastic cells. Bone. 2004 Sep; 35(3): 656-663.
    [38] Kheifets V, Bright R, Inagaki K, et al. Protein kinase C delta (deltaPKC)-annexin V interaction: a required step in deltaPKC translocation and function. J Biol Chem. 2006 Aug 11; 281(32): 23218-23226. Epub 2006 Jun 18.
    [1] Gerke V, Moss SE. Annexins: from structure to function. Physiol Rev. 2002 Apr; 82(2): 331-371.
    [2] Kahi CJ, Rex DK, Imperiale TF. Screening, surveillance, and primary prevention for colorectal cancer: a review of the recent literature. Gastroenterology. 2008; 135(2): 380-399.
    [3] Wilkinson N, Scott-Conner CE. Surgical therapy for colorectal adenocarcinoma. Gastroenterol Clin North Am. 2008; 37(1): 253-267, ix.
    [4] Wolpin BM, Mayer RJ. Systemic treatment of colorectal cancer. Gastroenterology. 2008; 134(5): 1296-1310.
    [5] Kasztelan-Szczerbińska B, Cichoz-Lach H, S?omka M. Colorectal cancer as a health care problem: evaluation of the current diagnostic options. Pol Arch Med Wewn. 2008; 118(4): 224-227.
    [6] Walsh JM, Terdiman JP. Colorectal cancer screening: scientific review. JAMA. 2003; 289(10): 1288-1296.
    [7] Boursi B, Arber N. Current and future clinical strategies in colon cancer prevention and the emerging role of chemoprevention. Curr Pharm Des. 2007; 13(22): 2274-2282.
    [8] Denlinger CS, Cohen SJ. Progress in the development of prognostic and predictive markers forgastrointestinal malignancies. Curr Treat Options Oncol. 2007; 8(5): 339-351.
    [9] Zlobec I, Lugli A. Prognostic and predictive factors in colorectal cancer. J Clin Pathol. 2008; 61(5): 561-569.
    [10] Hayes MJ, Moss SE. Annexins and disease. Biochem Biophys Res Commun 2004; 322: 1166-1170.
    [11] Gerke V, Moss SE. Annexins: from structure to function. Physiol Rev 2002; 82: 331-371.
    [12] Hawkins TE, Merrifield CJ, Moss SE. Calcium signaling and annexins. Cell Biochem Biophys. 2000; 33(3): 275-296.
    [13] Rescher U, Gerke V. Annexins--unique membrane binding proteins with diverse functions. J Cell Sci. 2004; 117(Pt 13): 2631-2639.
    [14] Futter CE, White IJ. Annexins and endocytosis. Traffic. 2007; 8(8): 951-958.
    [15] Mira JP, Dubois T, Oudinet JP, et al. Inhibition of cytosolic phospholipase A2 by annexin V in differentiated permeabilized HL-60 cells. Evidence of crucial importance of domain I type II Ca2+-binding site in the mechanism of inhibition. J Biol Chem. 1997; 272(16): 10474-10482.
    [16] Karube A, Shidara Y, Hayasaka K, et al. Suppression of calphobindin I (CPB I) production in carcinoma of uterine cervix and endometrium. Gynecol Oncol. 1995; 58(3): 295-300.
    [17] Mulla A, Christian HC, Solito E, et al. Expression, subcellular localization and phosphorylation status of annexins 1 and 5 in human pituitary adenomas and a growth hormone-secreting carcinoma. Clin Endocrinol (Oxf). 2004; 60(1): 107-119.
    [18] Guzmán-Aránguez A, Olmo N, Turnay J, et al. Differentiation of human colon adenocarcinoma cells alters the expression and intracellular localization of annexins A1, A2, and A5. J Cell Biochem. 2005; 94(1): 178-193.
    [19] Boersma HH, Kietselaer BL, Stolk LM, et al. Past, present, and future of annexin A5: from protein discovery to clinical applications. J Nucl Med. 2005; 46(12): 2035-2050.
    [20] Kenis H, Hofstra L, Reutelingsperger CP. Annexin A5: shifting from a diagnostic towards a therapeutic realm. Cell Mol Life Sci. 2007; 64(22): 285928-62.
    [21] Monceau V, Belikova Y, Kratassiouk G, et al. Externalization of endogenous annexin A5 participates in apoptosis of rat cardiomyocytes. Cardiovasc Res. 2004; 64(3): 496-506.
    [22] Cardó-Vila M, Arap W, Pasqualini R. Alpha v beta 5 integrin-dependent programmed cell death triggered by a peptide mimic of annexin V. Mol Cell. 2003; 11(5): 1151-1162.
    [23] Kheifets V, Bright R, Inagaki K, et al. Protein kinase C delta (deltaPKC)-annexin V interaction: a required step in deltaPKC translocation and function. J Biol Chem. 2006; 281(32): 23218-23226.
    [24] Yamaguchi T, Matsuda K, Sagiya Y, et al. p53R2-dependent pathway for DNA synthesis in a p53-regulated cell cycle checkpoint. Cancer Res. 2001; 61(22): 8256-8262.
    [1] Ouellette JR,Harboe-Schmidt JE,luthringer D,Brackert S,Silberman AW. Coloractal cancer metastasis presenting as a testicular mass:case report and review of the literature.Am Surg.2007Jan;73(1):79-81
    [2] Singh P, Wu H, Clark C, Annexin II binds progastrin and gastrin-like peptides, and mediates growth factor effects of autocrine and exogenous gastrins on colon cancer and intestinal epithelial cells. Oncogene. 2007 Jan 18; 26(3): 425-440. Epub 2006 Jul 10.
    [3] Bird NC, Mangnall D, Majeed AW. Biology of colorectal liver metastases: A review. J Surg Oncol. 2006 Jul 1; 94(1): 68-80.
    [4] Saletti P, Cavalli F. Metastatic colorectal cancer. Cancer Treat Rev. 2006 Nov; 32(7): 557-571. Epub 2006 Aug 28.
    [5] Takayama T, Miyanishi K, Hayashi T, et al. Colorectal cancer: genetics of development and metastasis. J Gastroenterol. 2006 Mar; 41(3): 185-192.
    [6] Rudmik LR, Magliocco AM. Molecular mechanisms of hepatic metastasis in colorectal cancer. J Surg Oncol. 2005 Dec 15; 92(4): 347-359.
    [7] Gerke V, Moss SE. Annexins: from structure to function. Physiol Rev. 2002 Apr; 82(2): 331-371.
    [8] Bandorowicz-Pikula J, Buchet R, Pikula S. Annexins as nucleotide-binding proteins: facts and speculations. Bioessays. 2001 Feb; 23(2): 170-178.
    [9] Seaton BA, Dedman JR. Annexins. Biometals. 1998 Dec; 11(4): 399-404.
    [10] Lim LH, Pervaiz S. Annexin 1: the new face of an old molecule. FASEB J. 2007 Apr; 21(4): 968-975. Epub 2007 Jan 10.
    [11] Terranova VP, DiFlorio R, Hujanen ES, et al. Laminin promotes rabbit neutrophil motility and attachment. J Clin Invest. 1986 Apr; 77(4): 1180-1186.
    [12] Terranova VP, Lyall RM. Chemotaxis of human gingival epithelial cells to laminin. A mechanism for epithelial cell apical migration. J Periodontol. 1986 May; 57(5): 311-317.
    [13] Liu W, Saint DA. Validation of a quantitative method for real time PCR kinetics. Biochem Biophys Res Commun. 2002 Jun 7; 294(2): 347-53.
    [14] Vasko VV,Saji M.Molecular mechanisms involved in differentiated thyroid cancer invasion and metastasis[J].Curr Opin Oncol,2007.19(1):11—17.
    [15] Erik w'r,Donald FN.Carcinoma invasion and metastasis:a role for epithelial—mesenchymal transition?[J]Cancer Res,2005,65(14):5591-5595.
    [16] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCRand the 2(-Delta Delta C(T)) Method. Methods. 2001 Dec; 25(4): 402-8.
    [17] Hattori T, Hoffman T, Hirata F. Differentiation of a histiocytic lymphoma cell line by lipomodulin, a phospholipase inhibitory protein. Biochem Biophys Res Commun. 1983 Mar 16; 111(2): 551-559.
    [18] Babbin BA, Lee WY, Parkos CA, et al. Annexin I regulates SKCO-15 cell invasion by signaling through formyl peptide receptors. J Biol Chem. 2006 Jul 14; 281(28): 19588-19599. Epub 2006 May 4.
    [19] Oh J, Rhee HJ, Kim S, et al. Annexin-I inhibits PMA-induced c-fos SRE activation by suppressing cytosolic phospholipase A2 signal. FEBS Lett. 2000 Jul 21; 477(3): 244-248.
    [20] Frohlich M, Motte P, Galvin K, et al. Enhanced expression of the protein kinase substrate p36 in human hepatocellular carcinoma. Mol Cell Biol. 1990 Jun; 10(6): 3216-3223.
    [21] Kumble KD, Hirota M, Pour PM, et al. Enhanced levels of annexins in pancreatic carcinoma cells of Syrian hamsters and their intrapancreatic allografts. Cancer Res. 1992 Jan 1; 52(1): 163-167.
    [22] Vishwanatha JK, Chiang Y, Kumble KD, Enhanced expression of annexin II in human pancreatic carcinoma cells and primary pancreatic cancers. Carcinogenesis. 1993 Dec; 14(12): 2575-2579.
    [23] Wang Y, Li ZF, He J, et al. Expression of the human phosphatases of regenerating liver (PRLs) in colonic adenocarcinoma and its correlation with lymph node metastasis. Int J Colorectal Dis. 2007 Apr 18; [Epub ahead of print]
    [24] Katayama M, Nakano H, Ishiuchi A, et al. Protein pattern difference in the colon cancer cell lines examined by two-dimensional differential in-gel electrophoresis and mass spectrometry. Surg Today. 2006; 36(12): 1085-1093. Epub 2006 Dec 25.
    [25] Sharma MR, Koltowski L, Ownbey RT, et al. Angiogenesis-associated protein annexin II in breast cancer: selective expression in invasive breast cancer and contribution to tumor invasion and progression. Exp Mol Pathol. 2006 Oct; 81(2): 146-156. Epub 2006 Apr 27.
    [26] Madoiwa S, Someya T, Hironaka M, et al. Annexin 2 and hemorrhagic disorder in vascular intimal carcinomatosis. Thromb Res. 2007; 119(2): 229-240. Epub 2006 Mar 9.
    [27] Sato H, Ogata H, De Luca LM. Annexin V inhibits the 12-O-tetradecanoylphorbol-13-acetate-induced activation of Ras/extracellular signal-regulated kinase (ERK) signaling pathway upstream of Shc in MCF-7 cells. Oncogene. 2000 Jun 8; 19(25): 2904-2912.
    [28] Sinicrope FA, Rego RL, Garrity-Park MM, et al. Alterations in cell proliferation and apoptosis in colon cancers with microsatellite instability. Int J Cancer. 2007 Mar 15; 120(6): 1232-1238.
    [1] Kamangar F,Dores GM,Anderson wf.Patterns of cancer incidence,mortality,and prevalence across five continents:defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 2006;24:2137-2150.
    [2] Eilstein D,Hedelin G,Schaffer P.Incidence of colorectal cancer in Bas-Rhin,trend and prediction in 2009.Bull cancer 2000;87:595-599.
    [3] Ji BT,Devesa SS,Chow WH,Jin F,Gao YT.Colorectal cancer incidence trends by subsite in urban shanghai,1992-1994.Cancer Epidemiol Biomarkers Prev 1998;7:661-666.
    [4] Alberts B.The cell as a collection of protein machines:preparing the next generation of molecular biologists.Cell 1998;92:291-294.
    [5] Anderson NL, Anderson NG.Proteome and proteomics:new technologies,new concepts,and new words.Electrophoresis 1998;19:1853-1861.
    [6] Appel RD,Palagi PM,Walther D,Vargas JR,Sanchez JC,Ravier F,Pasquali C,Hochstrasser DF.MelanieⅡ-a third-generation software package for analysis of two-dimensional electrophoresis images:I.Features and user interface.Electrophoresis 1997;18(15):2724-2734.
    [7] Gysi SP,Re.on Y,Frama B,et a1.Correlation between protein and mRNA abundancein yeast[J].Mol CellBiol,1999;19(3):1720-1730.
    [8] Mahon P,Dupree P.Quantitative and reproducible two-dimensional gel analysis using Phoretix 2D Full.Electrophoresis 2001;22:2075-2085.
    [9] Gygi SP,Corthais GL,Zhang Y,Rochon Y,Aebersold R.Evaluation of two-dim ensional gel electroDhoresis—based proteom e analysis technology.Proc Nat2 Acad Scf U S A 2000;97:9390-9395.
    [10] Blackstock WP,Weir MP.Proteomics:quantitative and physical mapping of cellular proteins.Trends Biotechnol,1999,17(3):121-127.
    [11] Anderson NL,Anderson NG.Proteome and proteomics:new technologies,new conepts,and new words.Electrophoresis,1998,19(11):1853-1861.
    [12] Wodicka L,Dong H ,Mittmann M ,et a1.Genome-wide expression monitoringin Saccharomyces cerevisiae.Nat Biotechnol,1997,15(13):1359-1367.
    [13] Ramsay G.DNA chips;state of-the art.Nature Biotech.1998,16(1):44.
    [14] Velculeseu VE.Zhang L,Vogelstein B,et a1.Serial analysis of gene exprssion.Science,1995,270(5235):484-487.
    [15] Perler FB,Olsen GJ,Adam E.Compilation and analysis of intein sequences.Nucleic Acids Res,1997,25(6):1087-1093.
    [16] Appel R D,Vargas J R,Palagi PM,et a1.MelanieⅡ-a third-generation software package for analysis of two dimensional electrophoresis image:Ⅱ.A1-gorithms.Electrophoresis,1997,18(15):2735-2748.
    [17] O‘Farrell PH .High resolution two-dim ensionalelectrophoresis of proteins.1 Bio2 Chem 1975:250:4007-4021.
    [18] Gorg A,W eiss W ,Dunn M I.Current two-dim ensional electrophoresis technologv for proteomi cs.Proteomics 2004:4:3665-3685.
    [19] Gygi SO,Corthals GL,Zhang Y,et a1.Evaluation of two-dimensional gel electrophoresis-based proteome analysis technology.Proc Natl Acad Sci U S A,2000,97(17):9390-9395.
    [20] W u SL,Hancock W S,Goodrich GG,Kunitake ST.An approach to the proteomic analysis of a breast cancer cell line(SKBR-3).Proteomics 2003;3:1037-1046 .
    [21] Alban A,David SO,Bjorkesten L,Andersson C,Sloge E,Lewis S,Currie I.A novel experimental design for comparative two-dimensional gel analysis:two-dimensional difference gel electrophoresis incorporating a pooled internal standard.Proteomics 2003;3:36-44.
    [22] Humphery-Smith I,Cordwell S J,Blackstock W P.Proteome research:Com-plamentarity and limitations with respect to the RNA and DNA worlds,Electrophoresis,1997,18(8):1217-1242.
    [23] Emmert-Buck MR,Bonner RF,Smith PD,Chuaqui RF,Zhuang Z,Goldstein SR,Weiss RA,Liotta LA.Laser capture microdissection.Science 1996;274:998-1001.
    [24] Hutchens TW , Yip TT . Protein interactions with surface-immobilized metal jLons :structure-dependent variations in afinity and binding capacitry with temperature and urea concentration.J Inorg Biochem 1991;42:105-118.
    [25] Chen YD,Zheng S,Yu JK,Hu X.Artificial neural,networks analysis of surface—enhanced laser desorption/ionization mass spectra of serum protein pattern distinguishes colorectal cancer from healthy poF,ulation.ain Cancer Res 2004;10:8380-8385.
    [26] Melle C,Ostefloh D,Ernst G,SchimmeI B,Bleu A,von Eggeling F.Identification of proteins from colorectal cancer tissue by two-dimensional gel electrophoresis and SELDI mass spectrometry.Int J Mol Med 2005;16:11-17.
    [27] Shiwa M ,Nishimura Y,Wakatabe R,Fukawa A,Arikuni H ,Ota H,Kato Y,Yamori T.Rapid discovery and identification of a tissue-specific tumor biomarker from 39 human cancer cell lines using the SELDI ProteinChip platform.Biochem Biophys Res Commun 20 03;309:18-25.
    [28] Albrethsen J,Bogebo R,Gammeltoft S,0lsen J,Winther B,Raskov H.Upregulated expression of human neutrophil peptides 1,2 and 3(HNP 1-3) in colon cancer serum and turnouts:a biomarker study.BMC Cancer2005;5:8.
    [29]王全晖,高春芳,王秀丽,赵光,李冬晖,许洋,马龙华.利用SELDI-TOF质谱技术分析大肠癌患者血清蛋白质谱的变化.中国病理生理杂志2005;21:1896-1900.
    [30] Stulik J,Koupilova K,Osterreieher J,et a1.Protein aboundance alterations in matched sets of macroscopicaily normal colon mucosa and colorectal carcinoma.Electrophoresis,1999,20(18):3638-3646.
    [31] Jungblut PR,Zimny Arndt U,Zeindl Eberhart E.et a1.Protemics in human disease:cancer,heart and infectious diseases Eletrophoresis.1999,20(10):2100-2l1O.
    [32] Chanrand P,DaGue BB,Pearsall RS.et a1.Profiling protein from azoxy methane-induced colon tumors at the moleculear level by matrix,assisted laser desorption/ionization mass spectrometry.Proteomics,2001;1(1O):1320-1326.
    [33]裴海平,朱红,曾亮,李宜雄.应用二维电泳和质谱技术筛选大肠癌与正常肠组织的差异表达蛋白中国普通外科杂志2005;14:748-752.
    [34] Cappello F,Di Stefano A,D’Anna SE,Donner CF,Zumlxlo G.Imm unopositivitv of heat shock protein 60 as a biomarker of bronchial carcinogenesis.Lancet 0no02 2005:6:816.
    [35] Barazi HO,Zhou L,Templeton NS,Krutzsch HC,Roberts DD.Identification of heat shock protein 60 as a molecular mediator of alpha 3 beta 1 integrin activation.Cancer RPs 2002;62:1541-1548.
    [36] Leonardi R,Pannone G,Magro G,Kudo Y,Takata T,Lo Muzio L.Differential expression of heat shock protein 27 in norm al oral mucosa , oral epithelial dysplasia and squamous cell carcinoma.Oncol Rep 2002;9:261-266.
    [37] Dressel R,Grzeszik C,Kreiss M ,Lindem ann D,Herrmann T,W alter L,Gunther E.Differential effect of acute and permanent heat shock protein 70 overexpression in tumor cells on lysability by cytotoxic T lymphocytes.Cancer Res 2003;63:8212-8220.
    [38] Zhao C,W an g E.Heat shock protein 90 suppresses tumor necrosis factor alpha induced apoptosis by preventing the cleavage of Bid in NIH3T3 fibroblasts.Cell Signal 2004;16:313-321.
    [39] Yamamoto K,Okam oto A,Isonishi S,0chiai K,Ohtake Y.Heat shock protein 27 was up-regulated in cisplatin resistan t human ovarian tumor cell line and associated with the cisplatin resistance.Cancer Lett 2001;168:173-181.
    [40] Srivastava P.Roles of heat-shock proteins in innate and adaptive immunity.Nat Rev Immunol 2002;2:185-194.
    [41]王树东,郑长青,刘明杰.不同类型热休克蛋白在大肠癌中的表达及其意义探讨.中国实用内科杂志2003;23:429-430.
    [42] Melle C,Bogumi l R,Ernst G,Schimm el B,Bleul A,von Eggeling F.Detection an d identification of heat,shock protein 10 as a biomarker in coloractal cancer by protein profiling.Proteomics 2006;6:2600-2608.
    [43] Cappello F Bellafiore M , Palma A t,David S,Marciano V,Bartolotta T,Sciume C,Modica G,Farina F,Zummo G,Bucchieri F.60KDa chaperonin(HSP60) is over-expressed during colorectal carcinogenesis.Eur J Histochem 2003;47:105-110.
    [44] Cappello F.Bellafiore M ,David S,Anzalone R,Zummo G.Ten kilodalton heat shock protein(HSP10)is overexpressed during carcinogenesis of large bowel and uterine exocervix.Cancer Lett 2003;196:35-41.
    [45] Coghlin C,Carpenter B,Dundas SR,Lawrie LC,Telfer C,Murray GI.Characterization and over-expression of chaperonin t-complex proteins in colorectal cancer.J PathoL 2006;210:351—357.
    [46]王少彬,陈俊辉,陈理明,黄杰雄,邱前程.热休克蛋白10在大肠癌的表达及其临床病理意义.中国肿瘤临床与康复2007;14:203—207.
    [47] Fearon ER,Vogelstein B.A genetic model for colorectal tumorigenesis.Cell 1990;61:759-767.
    [48] Bos JL,Fearon ER,HamiIton SR,Verlaan.De Vries M,van Boom JH,van der Eb AI,Vogelstein B.Prevalence of ras gene mutations in human colorectal cancers.Nature 1987;327:293-297.
    [49] Lin KM,Hollan der JM,Kao vY,Lin B,Macpherson L.Dillmarm W H.M yocvte protection by 10 kD heat shock protein(HsplO) involves the mobile loop and attenuation of the Ras GTP-ase pathway.FASEB J 2004;18:1004-1006.
    [50] Varela I,Cadinanos T,Pendas AM ,Gutierrez.Fernandez A,Folgueras AR,Sanchez LM ,Zhou Z,Rodriguez FJ,Stewart CL,Vega JA,TryggVason K,Freije JM,Lopez-otin C.Accelerated ageing in mice deficient in Zmpste24 protease is linked to p53 signalling activation.N 2005;437:564-568.
    [51] Attallah AM , Sakr HA , Isman H , Abdel-Hady el-SK , El-Dosokv I . An office-based immunodiagnostic assay for detecting urinary nuclear matrix protein 52 in patients with bladder cancer.BJU int 2005;96:334-339.
    [52] Lakshmanan Y,Subong EN,Partin AW .Differernial nuclear matrix protein expression in prostate cancers:correlation with pathologic stage.J Urol 1998;159:1354-1358.
    [53] Brunagel G,Vietmeier BN,Bauer AJ,Schoen RE,Getzenberg RH.Identification of nuclear matrix protein alterations associated with human colon cancer.Cancer Res 2002;62:2437-2442.
    [54]高静,王娅兰.大肠癌核基质蛋白变化与其生物学行为的关系.重庆医科大学学报2006;31:302-306.
    [55] Simpson R J,Connolly L M,Eddes J S,et a1.Proteomic analysis of the human colon carcinoma cell line(LIMI215):Development of a membrane protein database.Electrophoresis,2000,21(9):1707—1732.
    [56] Jungbhit PR,Zinmy Amdt U,Zeindl Eberhart E.et a1.Pmteomics in human disease:cancer,heart and infections disease[J].Elcctrophoresis.1999;20 :2100-2111.
    [57] Stulik J,Hernychova L,Porkertova S,eta1.Proteonle study of colorectal carcinogenesis.Electrophoresis,200l,22(14):3019-3025.
    [58] Stierum R,Gaspari M,Dommels Y,et a1.Proteome analysis reveals novel proteins associated with proliferation and differentiation of the colorectal cancer cell line Caco-2.Biochim Biophysa Acta,2003,1650(1-2):73-91.
    [59] Ahmed N,Oliva K,Wang Y,et a1.Proteomic profiling of proteins associated with urokinase plasminogen activator receptor in a colon cancer cell line using an antisense approach.Proteomics,2003,3(3):288-298.
    [60] Tan S,Scow TK,Liang RC.et a1.Proteome analysis of butyrateterated human colon cancer cells(HT-29).Int J Cnacer,2002.98(4):523-531.
    [61] Nakef A,Sabay N,Pisano M,et a1.Painting with a molecular brush:genomic/proteomic interfacing to define the drug action profile of novel solide-tumor selective anticancer agents.Cytometry,2002,47(1):72-79.
    [62] Simone NL.Remaley AT,Charboneau L,et a1.Sensitive Immunoassay of tissue cell proteins procured by laser capture microdissection.Am J Pathol,2000,156:445-452.
    [63] Shiwa M ,Nishinmra Y,Wakatahe R .et al. Rapid discovery and identification of a tissue-specific tumor biomarker from 39 human cancer cell lines using The SEI-DI Protein hip platform. BiochemBiophys Res Coninium,2003,309(1):I8-25.
    [64] John Roboz,IonghuaMa,Sung M.eta1.Protein profiles of serum in colon Cancer by SEIDL-TOF mass spectrometry[J].Proeomic:Poster Session AAcr,2002;32:123-124.
    [65] Gao chun fang,zhaog Guang,song guo yin,et a1.Discovery 0f biomarkers in the senum from colorectal; cancer by use of SDLDI-TOF mass spectrometry[J].Chin J Med,2003;26(11):658-661.
    [66] Ji H,Whitehead P,H,Reid GE,et a1.Two-dimensionald electrophoretic analysis of proteins expressed by normaland cancerous human crypts:application of mass spectromrtry to pepfide-mass fingerprinting[J].Electrophoresis,1994;15:391-405.
    [67] Ji H,Reid GE,Moritz RL,eta1.A two-dimensional gel database of human colon carcinoma proteins[J].Electrophoresis,1997;l8:605-613.Issaq HJ.The role of ceparation science in proteomics research.Electrophoresis,2001,22(17):3629-3638.
    [68] Rabilloud T.Tow-dimensional gel electrophoresis in proteomics:old,old fashioned,but it still climbs up the mountains.Proteomics,2002,2(1):3-10.
    [69] Shen Y,Tolic N,Zhao R,et a1.High-throughtput proteomics using highefficiency multiple-capillary liquid chromatography with on-line high performance ESI FTICE Mass spectrometry.Anal Chem,2001,73(13):3011-3021.
    [70] Simone N L,Remaley A T,Xharboneau L,et a1.Sensitive Immunoassay of Tissue Cell Proteins Procured by Laser Capture microdissection.Am J Pathol,2000,156(2):445-452.
    [71] Fuller AP,Palmer-Toy D,Erlander MG,et a1.Laser capture mierodissection and advanced molecular analysis of human breast cancer.J Mammary GIang Biol Neoplasia,2003,8(3):335-345.
    [72] Craven RA,Totty N,Hamden P,et a1.Laser capture mierodissection and two-dimensional polyacrylamide gel electrophoresis:evaluation of tissue preparation and sample limitations.Am J Pathol,2002,160(3):815-822.
    [73] Adam PJ,Boyb R,Tyson KL,et a1.Comprehensive proteomic analysis of breast cancer cell membrane reveals unique proteins with potential roles in clinical cancer.J Biol Chem,2003,278(8):6482-6489.

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