摘要
胰腺癌早期诊断困难,恶性程度高,手术切除率低,预后极差。在美国1997年死于胰腺癌的患者超过28000人,居癌症死亡的第5位。胰腺癌确诊后平均5年生存率低于5%。近年来随着各种治疗技术和诊断方法的发展,肿瘤的诊断率和生存率正在逐年提高,但胰腺癌的预后仍然很差。因此对于胰腺癌分子机制的研究,特别是寻找胰腺癌诊断和治疗的靶点及与胰腺癌调控相关的信号传导通路具有重要的意义。
随着人类基因计划的初步完成,蛋白质组学成为目前国际上的研究热点。与基因相比,蛋白质是人体的各种生命活动的直接执行者。通过对肿瘤组织与正常组织所表达的蛋白质的对比分析,可以找出其中的差异蛋白质。这些差异蛋白质通过其所在的信号传导通路调控肿瘤的发生和发展。
国际上出现的各种先进的实验技术和分析方法大大促进了蛋白质组学的发展。首先是高通量蛋白质筛选技术,如二维电泳、组织芯片、PBSⅡ/C型表面增强激光解吸离子化飞行时间质谱系统、pathway array分析等,这些方法的特点是可以同时检测大量的蛋白质,提高检测效率。高通量蛋白筛选技术能使我们在短时间内获得大量的数据,但传统的统计方法无法分析如此大量的数据。国际上通用的解决办法是将基因分析的技术(如软件和数据库等)用于分析蛋白质的功能和信号传导通路,可以有效的解决这一问题。基因芯片显著性分析(SAM)、基因芯片分析软件包(BRB-ArrayTools)、信号网络分析(Ingenuity Pathway Analysis, IPA)等技术目前常用于高通量蛋白质数据的分析。
使用表皮生长因子受体(EGFR)抑制剂治疗结直肠癌、非小细胞肺癌和乳腺癌的临床效果已经得到公认,目前EGFR抑制剂在胰腺癌治疗中的作用成为国内外研究的热点。西妥昔单抗和埃罗替尼已经被FDA批准用于临床治疗晚期胰腺癌。只有K-ras基因野生型的患者对EGFR抑制剂治疗敏感,因为K-ras基因突变会持续激活EGFR信号传导路的下游,从而使EGFR抑制剂的抑制作用不能向下传递。在这一过程中EGFR通路及其它信号传导通路的蛋白质表达情况国内外未见相关报道。
我们采用pathway array分析方法,筛选出胰腺癌组织和正常胰腺组织之间的差异蛋白,通过SAM、BRB-ArrayTools、IPA等统计分析软件,确定了12个有统计学意义的差异蛋白及其主要涉及的信号传导通路。同时测定了胰腺癌标本的K-ras基因突变情况,并对其对蛋白质表达的影响进行了分析。我们也结合标本的临床资料,探讨了胰腺癌患者蛋白质表达与年龄、性别、种族和淋巴结转移情况之间的关系。
Purpose:
Pancreatic cancer is the fourth leading death of cancer among men and women in the United States The overall survival rate is less than 1% at 5 years with most patients dying within one year[9]. The lack of diagnosis early method and effective treatment is the main problem to pancreatic cancer. We aim to identify protein signatures of pancreatic cancer for diagnosis and therapy.
Materials and Method:
we employed protein pathway array method to assess the protein expression of 11 paires pancreatic cancer samples,5 pancreatic samples and 2 normal pancreatic samples. Next, we use high resolution melting (HRM) curve analysis for KRAS mutation of all pancreatic cancer samples. In addition, Pathways associated with these expression changes were identified using the Ingenuity Pathway Analysis tool.
Results and Conclusion:
8 proteins were upregulated and 4 proteins were downregulated in pancreatic cancer compared with and normal tissues. KRAS mutations were identified in 9 of 15 tumor tissues (60%). p-PKCα/βⅡin K-ras positive upregulation. diffirent express protein mainly affects cell cycle, cellar Growth and Proliferation, Tissue Morphology network functions.IPA result demonstrated the proteins of our study is correlation with lots of signaling pathway.
Eight proteins (FAS, cdk2, cdc2 p34, EGFR, Akt, Vimentin, E-cadherin ICAM-1) were up-regulated and four protein (cyclin B1, Cdc25B, H-Ras, HIF-3α,) were down-regulated in pancreatic cancer compared with normal tissues.these differentially expressed proteins affect cell proliferation, cell cycle and apoptosis ect.these proteins can be used to early diagnosis and therapeutic target.
引文
[1]Jemal A, Siegel R, Ward E, et al. Cancer statistics,2008. CA Cancer J Clin,2008,58(2):71-96.
[2]病理生理学(第七版):11.
[3]Stolte M. The new Vienna classification of epithelial neoplasia of the gastro intestinal tract:advantages and disadvantages. Virchow sArch,2003,442: 992106.
[4]Takaori K, Hruban RH, Maitra A, et al. Pancreatic intraepithelial neoplasia. Pancreas,2004,28:257-262.
[5]Iacobuzio-Donahue CA, Hruhan RH. Gene expression in neoplasms of pancreas:applications to diagnostic pathology. Adv Anat Pathol, 2003, 3:125-134.
[6]Hruban RH, Wilentz RE, Kern SE Genetic progression in the pancreatic ducts. Am. J. Pathol.,156:1821-1825,2000.
[7]Eberhard DA, Johnson BE, Amler LC, et al.(2005) Mutations in the epidermal growth factor receptor and in KRAS are predictive and prognostic indicators in patients with non-small-cell lung cancer treated with chemotherapy alone and in combination with erlotinib. J Clin Oncol 23:5900-5909.
[8]Karapetis CS, Khambata-Ford S, Jonker DJ, O'Callaghan CJ, Tu D, Tebbutt NC, Simes RJ, Chalchal H, Shapiro JD, Robitaille S et al. (2008) K-ras mutations and benefit from cetuximab in advanced colorectal cancer. N Engl J Med 359,1757-1765. CrossRef,PubMed,ChemPort,Web of Science(?) Times Cited:1855Amado RG, Wolf M, Peeters M, Van Cutsem E, Siena S, Freeman DJ, Juan T, Sikorski R, Suggs S, Radinsky R et al. (2008) Wild-type KRAS is required for panitumumab efficacy in patients with metastatic colorectal cancer. J Clin Oncol 26,1626-1634. CrossRef,PubMed,ChemPort,Web of Science(?) Times Cited:293.
[9]Eberhard DA, Johnson BE, Amler LC, et al.(2005) Mutations in the epidermal growth factor receptor and in KRAS are predictive and prognostic indicators in patients with non-small-cell lung cancer treated with chemotherapy alone and in combination with erlotinib. J Clin Oncol 23:5900-5909.
[10]Slamon DJ, Clark GM, Wong SG,et al:Human breast cancer: Correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235:177-182,1987.
[11]Dati C, Muraca R, Tazartes O et al.c-erb-2 and ras expreassion levels in breast cancer are correlated and show a cooperative association with unfavorale clinical outcome. Int J Cancer, 1991; 47 (6):833.
[12]ImaiY, TsurutaniN, OdaH, et al p16INK4 genemutations are re latively frequent in ampullary carc inom as[J]. Jpn J Cancer Res,1997,88(10): 941-946.
[13]Attri J, Sr in ivasan R, M ajumdar S, et a.l A lte rations o f tumo r suppresso r gene p16INK4a in panc reatic ducta 1 carcinom a[J]. BMC G astroenterol,2005,5(28):22.
[14]Shiraham a S, Matsuyosh i J, Yamamo to S, et a.l A Iteration of the K-ras and p16/CDKN2 gene in endo crine pancreatic carcinom as[J]. N ippon R insho,2004,62 (5):889-893.
[15]Jin X, Ngugend D, Zhang WW, e t a.l Ce 11 cyc le arrest and inhibition of tum or ce 11 pro liferation by the p16 INK4 gene med ia ted by an adenov irus vector[J]. Cancer Res,1995,55(15):3250-3253.
[16]Datto M, Wang XF. The Smads:transcriptional regulation and mouse models. Cytokine Growth Factor Rev 2000; 11:37-48.
[17]Proteomic profiling in pancreatic cancer with and without lymph node metastasis.
[18]Proteomic and Tissue Array Profiling Identifies Elevated Hypoxia-Regulated Proteins in Pancreatic Ductal Adenocarcinoma.
[19]Expression of Ezrin and Phosphorylated Ezrin (pEzrin) in Pancreatic Ductal Adenocarcinoma.
[20]Proteomic analysis of pancreatic ductal adenocarcinoma compared with normal adjacent pancreatic tissue and pancreatic benign cystadenoma.
[21]Parkin DM, Bray F, Ferlay J, Pisani P. Global cancer statistics,2002. CA Cancer J Clin 2005; 55:74-108.
[22]Agency for Healthcare Research and Quality (AHRQ).Screening for Pancreatic Cancer. Recommendation Statement. U.S.Department of Health & Human Services. Washington, DC, USA,2004.
[23]Brentnall TA, Bronner MP, Byrd DR, Haggitt RC, Kimmey MB.Early diagnosis and treatment of pancreatic dysplasia in patients with a family history of pancreatic cancer. Ann Intern Med 1999; 131:247-55.
[24]Canto MI, Goggins M, Yeo CJ, Griffin C, Axilbund JE, Brune K, et al. Screening for pancreatic neoplasia in high-risk individuals:an EUS-based approach. Clin Gastroenterol Hepatol 2004; 2:606-21.
[25]Canto MI. Screening for pancreatic neoplasia in high-risk individuals: who, what, when, how? Clin Gastroenterol Hepatol 2005; 3(7 Suppl. 1):S46-8.
[26]Wang SJ, Gao Y, Chen H, Kong R, Jiang HC, Pan SH, Xue DB, Bai XW,Sun B (2010) Dihydroartemisinin inactivates NF-kappaB and potentiates the anti-tumor effect of gemcitabine on pancreatic cancer both in vitro and in vivo. Cancer Lett 293(1):99-108.
[27]Yasumoto K, Koizumi K, Kawashima A, Saitoh Y, Arita Y, Shinohara K, Minami T, Nakayama T, Sakurai H, Takahashi Y, Yoshie O, Saiki I (2006) Role of the CXCL12/CXCR4 axis in peritoneal carcinomatosis of gastric cancer. Cancer Res 66(4):2181-2187.
[28]Yokoi K, Fidler IJ (2004) Hypoxia increases resistance of human pancreatic cancer cells to apoptosis induced by gemcitabine. Clin Cancer Res 10(7):2299-2306.
[29]Wang Z, Li Y, Kong D, Banerjee S, Ahmad A, Azmi AS, Ali S, Abbruzzese JL, Gallick GE, Sarkar FH (2009) Acquisition of epithelial-mesenchymal transition phenotype of gemcitabine-resistant pancreatic cancer cells is linked with activation of the notch signaling pathway. Cancer Res 69(6):2400-2407.
[30]Wong HH, Lemoine NR (2009) Pancreatic cancer:molecular pathogenesis and new therapeutic targets. Nat Rev Gastroenterol Hepatol 6(7):412-422.
[31]Tachikui H, Navet AF and Ozawa M:Identification of theCa(2+)-binding domains in reticulocalbin, an endoplasmicreticulum resident Ca(2+)-binding protein with multiple EF-hand motifs. J Biochem 121:145-149,1997.
[32]Tong A, Zhang H, Li Z, Gou L, Wang Z, Wei H, Tang M, Liang S, Chen L, Huang C and Wei Y:Proteomic analysis of liver cancer cells treated with suberonylanilide hydroxamic acid. Cancer Chemother Pharmacol 61: 791-802,2008.
[33]Morgan J, Hoekstra AV, Chapman-Davis E, Hardt JL, Kim JJ and Buttin BM:Synuclein-gamma (SNCG) may be a novel prognostic biomarker in uterine papillary serous carcinoma. Gynecol Oncol 114:293-298,2009.
[34]Bram RJ, Hung DT, Martin PK, Schreiber SL and Crabtree GR: Identification of the immunophilins capable of mediating inhibition of signal transduction by cyclosporin A and FK506:roles of calcineurin binding and cellular location. Mol Cell Biol 13:4760-4769,1993.
[35]Colgan J, Asmal M, Yu B and Luban J:Cyclophilin A-deficient mice are resistant to immunosuppression by cyclosporine. J Immunol 174: 6030-6038,2005.
[36]Zhu D, Cardenas ME and Heitman J:Calcineurin mutants render T lymphocytes resistant to cyclosporin A. Mol Pharmacol 50:506-511,1996.
[37]Singh VK, Zhou Y, Marsh JA, Uversky VN, Forman-Kay JD, Liu J and Jia Z:Synuclein-gamma targeting peptide inhibitor that enhances sensitivity of breast cancer cells to antimicrotubule drugs. Cancer Res 67: 626-633,2007.
[1]Buchholz, Maltel; Gress, Thomas M. Molecular changes in pancreatic cancer. olume 9, Number 10, October 2009, pp.1487-1497(11).
[2]Bos J (1989). "ras oncogenes in human cancer:a review". Cancer Res 49(17):4682-9.
[3]Pathologic Basis of Disease 8th ed..2010. p.282.
[4]Kras regulatory elements and exon 4A determine mutation specificity in lung cancer.
[5]M. Macaluso, G. Russo, C. Cinti, V. Bazan, N. Gebbia, A. Russo. Ras family genes:An interesting link between cell cycle and cancer. Journal of Cellular Physiology Volume 192, Issue 2, pages 125-130, August 2002.
[6]Krejs GJ. Pancreatic cancer:epidemiology and risk factors. Dig Dis. 2010;28(2):355-8. Epub 2010 Sep 1.
[7]Maisonneuve P, Lowenfels AB. Epidemiology of pancreatic cancer:an update. Dig Dis.2010;28(4-5):645-56. Epub 2010 Nov 18.
[8]Ahmedin Jemal, DVM, PhD, Rebecca Siegel, MPH, Elizabeth Ward, PhD, Yongping Hao, PhD, Jiaquan Xu, MD*, Taylor Murray and Michael J. Thun, MD, MS. Cancer Statistics,2008. CA Cancer J Clin 2008; 58:71-96
[9]National Cancer Institute. Pancreatic Cancer Treatment (PDQ(?)). Accessed 04 Feb 2010.
[10]Hutter, G., and Sinha, P. (2001) Proteomics for studying cancer cells and the development of chemoresistance. Proteomics 1,1233-1248
[11]Lohr, M., and Faissner, R. (2004) Proteomics in pancreatic disease. Pancreatology.4,67-75.
[12]Wulfkuhle, J. D., Liotta, L. A., and Petricoin, E. F. (2003) Proteomic applications for the early detection of cancer. Nat. Rev. Cancer 3,267-275.
[13]Baron A, Moore PS, Scarpa A:DNA array/microarrays in oncological research with focus on pancreatic cancer. Adv Clin Path 2001; 5:115-120.
[14]Sato N, Fukushima N, Maitra A, Iacobuzio-Donahue CA, van Heek NT, Cameron JL, Yeo CJ, Hruban RH, Goggins M:Gene expression profiling identifies genes associated with invasive intraductal papillary mucinous neoplasms of the pancreas. Am J Pathol 2004; 164:903-914.
[15]J emal A, Siegel R, Ward E, et al. Cancer st at ist ics,2009[J]. CA Cancer J Clin,2009,59(4):225□249.
[16]Geer RJ, Brennan MF. Prognostic indicators for survival after resection of pancreatic adenocarcinoma. Am J Surg 1993; 165:68-72.
[17]annual review of pathology:mechanisms of disease.2008:3:157-188.
[18]Jeffery DA, Bogyo M Chemical proteomics and its application to drug discovery. (2003) Curr OpinBiotechnol 14(1):87-95.
[19]Jones S, Zhang X, Parsons DW, Lin JC, Leary RJ, Angenendt P, Mankoo P, Carter H, Kamiyama H, Jimeno A,Hong SM, Fu B, Lin MT, Calhoun ES, Kamiyama M,Walter K, Nikolskaya T, Nikolsky Y, Hartigan J, Smith DR,Hidalgo M, Leach SD, Klein AP, Jaffee EM, Goggins M,Maitra A, Iacobuzio-Donahue C, Eshleman JR, Kern SE,Hruban RH, Karchin R, Papadopoulos N, Parmigiani G,Vogelstein B, Velculescu VE, Kinzler KW. Core signaling pathways in human pancreatic cancers revealed by global genomic analyses. Science,2008,321(5897):1801-1806.
[20]Gill, G.N., et al. Monoclonal anti-epidermal growth factor receptor antibodies which are inhibitors of epidermal growth factor binding and antagonists of epidermal growth factor binding and antagonists of epidermal growth factor-stimulated tyrosine protein kinase activity. J. Biol. Chem.1984, 259(12),7755-7560.
[21]SWOG S0502:Phase Ⅲ Randomized Study of Gemcitabine With Versus Without Cetuximab as First-Line Therapy in Patients With Locally Advanced Unrestable or Metastatic Adenocarcinoma of the Pancreas. Clin. Adv. Hematol. Oncol.2004,2 (4),201-252.
[22]Goldstein, N.I., et al. Biological efficacy of a chimeric antibody to the epidermal growth factor receptor in a human tumor xenograft model. Clin. Cancer Res.1995,1 (11),1311-1318.
[23]Sato, J.D., et al. Biological effects in vitro of monoclonal antibodies to human epidermal growth factor receptors. Mol. Biol. Med.1983,1 (5), 511-529.
[24]Alberts, S.R.; Sinicrope, F.A.; Grothey, A. N0147:a randomized phase III trial of oxaliplatin plus 5-fluorouracil/leucovorin with or without cetuximab after curative resection of stage Ⅲ colon cancer. Clin. Colorectal Cancer.2005, 5 (3),211-213.
[25]Burtness, B., et al. Phase Ⅲ randomized trial of cisplatin plus placebo compared with cisplatin plus cetuximab in metastatic/recurrent head and neck cancer:an Eastern Cooperative Oncology Group study. J. Clin. Oncol.2005,23 (34),8646-8654.
[26]Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. CancerStatistics, 2009. CA Cancer J Clin 2009; 59:225-49.
[27]Grote VA, Becker S, Kaaks R. Diabetes mellitus type 2:an independent risk factor for cancer? Exp Clin Endocrinol Diabetes 2010; 118:4-8.
[28]Tsugane S, Inoue M. Insulin resistance and cancer:epidemiological evidence. Cancer Sci 2010; 101:1073-9.
[29]Olowokure O, Beg MS, Ali S, Tandra A, Safa M, Havlin K, Ahmad SA. Is diabetes mellitus (DM) associated with worse outcomes in pancreatic cancer (PC)? J Clin Oncol 2010; 28(15Suppl):4114.
[30]Shama MA,Tanaka M, Curley SA, Abbruzzese JL, Li D. Association of diabetes with perineural invasion and overall survival in surgically resected patients with pancreatic cancer. J Clin Oncol 2010; 28(15 Suppl):4117.
[31]Saruc M, Karaarslan M, Rasa K, Saygili O, Ince U, Baysal C, et al. Pancreatic cancer and glucose metabolism. Turk J Gastroenterol 2009; 20:257-60.
[32]Saruc M, Pour PM. Diabetes and its relationship to pancreatic carcinoma. Pancreas 2003; 26:381-7.
[33]Garg P, Gupta, R. Pancreatic cancer and diabetes:is diabetes a risk factor or the outcome? J Clin Oncol 2007; 25(18 Suppl):15123.
[34]Busaidy N, Yazbeck C, Shah P, Evans D, Li D, Geraci JM, Weiser M. Survival of resectable pancreatic cancer patients with diabetes. J Clin Oncol 2006; 24(18 Suppl):4098.
[35]Feng Y, Busaidy N, Yeung S. Differential impact of antidiabetic treatment on pancreatic cancer cell growth in cell culture conditions mimicking different stages in the natural history of diabetes mellitus type 2. J Clin Oncol 2008;26:(15 Suppl):4640.
[36]Jemal A, Siegel R, Ward E, Hao Y, Xu J, Thun MJ. Cancer Statistics, 2009. CA Cancer J Clin 2009; 59:225-49.
[37]G.V. Glinsky, A.B. Glinskii, A.J. Stephenson, R.M. Hoffman and W.L Gerald, Gene expression profiling predicts clinical outcome of prostate cancer 2, J Clin Invest 113 (2004), pp.913-923.
[38]G. Narla, A. Difeo and H.L. Reeves et al., A germline DNA polymorphism enhances alternative splicing of the KLF6 tumor suppressor gene and is associated with increased prostate cancer risk, Cancer Res 65 (2005), pp.1213-1222.
[39]A.C. Lockhart, M.L. Rothenberg and J.D. Berlin, Treatment for pancreatic cancer:current therapy and continued progress, Gastroenterology 128 (2005), pp.1642-1654.
[40]H.A. Burris III, M.J. Moore and J. Andersen et al., Improvements in survival and clinical benefit with gemcitabine as first-line therapy for patients with advanced pancreas cancer:a randomized trial, J Clin Oncol 15 (1997), pp. 2403-2413.
[41]S. Yea, G. Narla and X. Zhao et al., Ras promotes growth by alternative splicing-mediated inactivation of the KLF6 tumor suppressor in hepatocellular carcinoma, Gastroenterology 134 (2008), pp.1521-1531.
[42]G. Narla, A. Difeo and S. Yao et al., Targeted inhibition of the KLF6 splice variant, KLF6 SV1, suppresses prostate cancer cell growth and spread 2, Cancer Res 65 (2005), pp.5761-5768.
[43]G. Schneider, J.T. Siveke, F. Eckel and R.M. Schmid, Pancreatic cancer:basic and clinical aspects, Gastroenterology 128 (2005), pp. 1606-1625.
[44]G. Garcea, C.P. Neal, C.J. Pattenden, W.P. Steward and D.P. Berry, Molecular prognostic markers in pancreatic cancer:a systematic review, Eur J Cancer 41 (2005), pp.2213-2236.
[45]Mori-Iwamoto S, Kuramitsu Y, Ryozawa S, Mikuriya K, Fujimoto M, Maehara S, Maehara Y, Okita K, Nakamura K and Sakaida I:Proteomics finding heat-shock protein 27 as a biomarker for resistance of pancreatic cancer cells to gemcitabine. Int J Oncol 31:1345-1350,2007.
[46]Kuramitsu Y, Miyamoto H, Tanaka T, Zhang X, Fujimoto M, Ueda K, Tanaka T, Hamano K and Nakamura K:Proteomic differential display analysis identified up-regulated astrocytic phsphoprotein PEA-15 in human malignant pleural mesothelioma cell lines. Proteomics 9:5078-5089,2009.
[47]Tanaka T, Kuramitsu Y, Fujimoto M, Naito S, Oka M and Nakamura K:Down-regulation of two isoforms of ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCH-L1) correlates with high metastatic potentials of human SN12C renal cell carcinoma cell clones. Electrophoresis 29:2651-2659,2008.
[48]Takashima M, Kuramitsu Y, Yokoyama Y, Iizuka N, Harada T, Fujimoto M, Sakaida I, Okita K, Oka M and Nakamura K:Proteomic analysis of autoantibodies in patients with hepatocellular carcinoma. Proteomics 6: 3894-3900,2006.
[49]Maehara S, Tanaka S, Shimada M, Shirabe K, Saito Y, Takahashi K and Maehara Y:Selenoprotein P, as a predictor for evaluating gemcitabine resistance in human pancreatic cancer cells. Int J Cancer 112:184-189,2004.
[50]Alexios S Strimpakos, Konstantinos N Syrigos, Muhammad W Saif, Translational Research in Pancreatic Cancer, J Pancreas (Online) 2010 Mar 5; 11(2):124-127
[51]Nagata S, Suda T. Immunol Today,1995,16:39-42.
[52]Itoch N, Nagata S. J Biol Chem,1993,268:10932-10938.
[53]Koepp DM, Harper JW, Elledge SJ. How the cyclin become a cyclin: regulated proteolysis in the cell cycle [J]. Cell,1999,97(4):431-434
[54]Liu E, Li X, Yan F, et al. Cyclin-dependent kinases phosphorylate human Cdtl and induce its degradation [J]. J Biol Chem,2004,279(17): 17283-17288
[55]Sullivan SM, Holyoak T. Enzymes with lid-gated active sites must operate by an induced fit mechanism instead of conformational selection[J]. Proc Natl Acad Sci USA,2008,105(37):13829-1383
[56]Okazaki K, Takada S. Dynamic energy landscape view of coupled binding and protein conformational change:Induced-fit versus population-shift mechanisms [J]. Proc Natl Acad Sci USA,2008,105(32):11182-11187
[57]Yang LW, Eyal E, Bahar I, et al. Principal component analysis of native ensembles of biomolecular structures (PCA_NEST):Insights into functional dynamics[J]. Bioinformatics,2009,25(5):606-614
[58]Huang SY, Zou X. Efficient molecular docking of NMR structures: Application to HIV-1 protease[J]. Protein Sci,2007,16(1):43-51
[59]Friess H,Wang L,Zhu Z, et al. Growth factor receptors are diff erent ially expressed in can cers of the papilla of vat er and pancreas. Ann Surg,1999, 230(6):767-775.
[60]Watanabe S,sato K,Okazaki Y,et al.Activation of PI3K-AKT pathway in oral epithelial dysplasia and early cancer of tongue.Bull Tokyo Dent Coll,2009,50(3):125-133.
[61]Al-Saad S, Donnem T,Al-Shibli K, et al.Diverse prognostic roles of Akt isoforms, PTEN and PI3k in tumor epithelial cells and stromal comp artment in non-small cell lung cancer.Anticancer Res,2009,29(10):4175-4183.
[62]Hoekstra AV, Sefton EC,Berry E,et al.Progestins activate the AKT pathway in leiomyoma cells and promote survival.J Clin Enadocrinol Metab,2009,94(5):1768-1774.
[63]Yamamoto S.Tomita Y,Hoshida Y,etal.Prognostic significance of activated Akt expression in pancreat ic ductal adenocarcinoma.Clin Cancer Res,2004,10(8):2846-2850.
[64]Chadha KS Khoury T,Yu J,et al.Activated Akt and Erk expression and survival after surgery in pancreatic carcinoma Ann Surg Oncol,2006,13 (7):933-939.
[65]Pham NA,Tsao MS,Cao P,et al.Dissociation of gemcitabine sensitivity and protein kinase B signaling in pancreatic ductal adenocarcinoma models. Pancreas,2007,35(3):e 16-26.
[66]Covello KL, SimonMC, Keith B. Targeted replacement of hypoxia-inducible factor-1 alpha by a hypoxia-inducible factor-2alpha knock-in allele promotes tumor growth[J]. Cancer Res,2005,65:2277-2286.
[67]Tian H, McKnight SL, RussellDW. Endothelial PAS domain protein 1(EPAS1),a transcription factor selectively expressed in endothelial cells[J]. Genes Dev,1997,11:72-82.
[68]Hara S, Hamada J, Kobayashi C, et al. Expression and characterization of hypoxia-inducible factor (HIF)-3alpha in human kidney: supp ression of H IF-mediated gene expression by HIF-3alpha[J]. Biochem Biophys Res Commun,2001,287:808-813.
[69]Semenza GL,Wang GL. A nuclear factor induced by hypoxia via de novo protein synthesis binds to the human erythropoietin gene enhancer at a site required for transcrip tionalactivation[J]. Mol Cell Biol,1992,12: 5447.5454.
[70]Gu YZ, Moran SM, Hogenesch JB, et al Molecular characterization and chromosomal localization of a third alpha-class hypoxia inducible factor subunit, HIF3alpha[J]. Gene Expr,1998,7:205-213.
[71]杨盛力,陈孝平,张万广,刘利平,梁慧芳,任利,占大钱. HIF-3a在肝癌组织中的表达及临床意义[J]肝胆外科杂志,2008,3:229-231.
[72]Facina G, Lopes- Costa P V, Dos Santos A R, et al.Immunohistochemical expression of E- cadherin in sclerosing adenosis, ductal carcinoma in situ and invasive ductal carcinoma of the breast[J].Diagn Cytopathol,2010,38 (11):235-238.
[73]Sasaki K, Natsugoe S, Ishigami S, et al.Significance of Twist expression and its association with E- cadherin in esophageal squamous cell carcinoma[J]. J Exp Clin Cancer Res,2009,21:158.
[74]张军,姜希宏,马榕,等.乳腺癌E-cadherin的表达与术野脱落癌细胞关系的研究[J].山东医科大学学报,2000,38(1):53
[75]蔡建春,江绍基,陈治平,等.钙依赖细胞粘附分子与胃癌的生长方式和分化程度的关系[J].中华病理学杂志,1994,23(3):132
[76]er rara N, Henzed WJ. Pituitar y follicular cells secrete a novel hepar in-binding grow th factor specific for vascular endot helial cells [J]. Bio chem Biophys Res Commun,1989,161(2):851
[77]Senger DR, Connolly DT, Van-de-water L, et al. Purification and NH2-terminal amino acid sequence of guinea pig tumor secreted vascular permeability facto r [J]. Cancer Res,1990,50(6):1774
[78]Kuehn R, Lelkes PI, Bloechle C, et al. Angiogenesis, angiogenic growth factors, and cell adhesion molecules are upregulated in chronic pancreatic diseases:angiogenesis in chronic pancreatitis and in pancreatic cancer. Pancreas,1999,18:96-103.
[79]Wang J, Springer TA. Structural specializations of immunoglobulin superfamily members for adhesion to integrins and viruses. Immunol Rev 1998; 163:197-215
[80]Kalogeris TJ, Laroux FS, Cockrell A, Ichikawa H, Okayama N, Phifer TJ, Alexander JS, Grisham MB. Effect of selective proteasome inhibitors on TNF-induced activation of primary and transformed endothelial cells. Am J Physiol 1999; 276:C856-864
[81]Lyons PD, Benveniste EN. Cleavage of membrane-associated ICAM-1 from astrocytes:involvement of a metalloprotease. Glia 1998; 22: 103-112
[82]Wakatsuki T, Kimura K, Kimura F, Shinomiya N, Ohtsubo M, Ishizawa M, Yamamoto M. A distinct mRNA encoding a soluble form of ICAM-1 molecule expressed in human tissues. Cell Adhes Commun 1995; 3: 283-292