Adrenomedullin expression in epithelial ovarian cancers and promotes HO8910 cell migration associated with upregulating integrin α5β1 and phosphorylating FAK and paxillin

Journal of Experimental & Clinical Cancer Research

Yi Zhang⁴  Fang Wen⁴  Xiaoli Zhang¹  Zhuang Han³  Yuan Miao¹  Siyang Zhang²  Boya Deng⁴ 

[1] Department of Pathology, The First Hospital and College of Basic Medical Sciences of China Medical University, Shenyang 110001, China;Center of Laboratory Technology and Experimental Medicine, China Medical University, Shenyang 110001, China;Department of Orthopaedics, The First Hospital of China Medical University, Shenyang 110001, China;Department of Gynecology, The First Hospital of China Medical University, Shenyang 110001, China

关键词: Paxillin;    FAK;    Integrin α5β1;    Migration;    Progression;    Carcinogenesis;    AM;    Epithelial ovarian cancer;   

Others  :  826764
DOI  :  10.1186/1756-9966-31-19

 received in 2012-01-17, accepted in 2012-03-09,  发布年份 2012

【 摘 要 】

Background

Epithelial ovarian cancer (EOC) is one of the leading causes of cancer deaths in women worldwide. Adrenomedullin (AM) is a multifunctional peptide which presents in various kinds of tumors.

Methods

In this study, we characterized the expression and function of AM in epithelial ovarian cancer using immunohistochemistry staining. Exogenous AM and small interfering RNA (siRNA) specific for AM receptor CRLR were treated to EOC cell line HO8910. Wound healing assay and flow cytometry were used to measure the migration ability and expression of integrin α5 of HO8910 cells after above treatments. Western blot was used to examine the phosphorylation of FAK and paxillin.

Results

We found that patients with high AM expression showed a higher incidence of metastasis, larger residual size of tumors after cytoreduction and shorter disease-free and overall survival time. Exogenous AM induced ovarian cancer cell migration in time- and dose- dependent manners. AM upregulated the expression of integrin α5 and phosphorylation of FAK, paxillin as well.

Conclusions

Our results suggested that AM contributed to the progression of EOC and had additional roles in EOC cell migration by activating the integrin α5β1 signaling pathway. Therefore, we presumed that AM could be a potential molecular therapeutic target for ovarian carcinoma.

【 授权许可】

   
2012 Deng et al; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Permuth-Wey J, Sellers TA: Epidemiology of ovarian cancer. Methods Mol Biology (Clifton, NJ) 2009, 472:413-437.
• [2]Vang R, Shih I-M, Kurman RJ: Ovarian Low-grade and High-grade Serous Carcinoma Pathogenesis, Clinicopathologic and Molecular Biologic Features, and Diagnostic Problems. Adv Anat Pathol 2009, 16(5):267-282.
• [3]Lengyel E: Ovarian cancer development and metastasis. Am J Pathol 2010, 177(3):1053-1064.
• [4]Kitamura K, Kangawa K, Kawamoto M, Ichiki Y, Nakamura S, Matsuo H, Eto T: Adrenomedullin: a novel hypotensive peptide isolated from human pheochromocytoma. Biochem Biophys Res Commun 1993, 192(2):553-560.
• [5]Wimalawansa SJ: Amylin, calcitonin gene-related peptide, calcitonin, and adrenomedullin: A peptide superfamily. Crit Rev Neurobiol 1997, 11(2-3):167-239.
• [6]Zudaire E, Martinez A, Cuttitta F: Adrenomedullin and cancer. Regul Pept 2003, 112(1- 3):175-183.
• [7]Abasolo I, Montuenga LM, Calvo A: Adremomedullin prevents apoptosis in prostate cancer cells. Regul Pept 2006, 133(1-3):115-122.
• [8]Martinez A, Vos M, Guedez L, Kaur G, Chen Z, Garayoa M, Pio R, Moody T, Stetler-Stevenson WG, Kleinman HK, et al.: The effects of adrenomedullin overexpression in breast tumor cells. J Natl Cancer Inst 2002, 94(16):1226-1237.
• [9]Hata K, Takebayashi Y, Akiba S, Fujiwaki R, Iida K, Nakayama K, Nakayama S, Fukumoto M, Miyazaki K: Expression of the adrenomedullin gene in epithelial ovarian cancer. Mol Hum Reprod 2000, 6(10):867-872.
• [10]Miller MJ, Martinez A, Unsworth EJ, Thiele CJ, Moody TW, Elsasser T, Cuttitta F: Adrenomedullin expression in human tumor cell lines. Its potential role as an autocrine growth factor. J Biol Chem 1996, 271(38):23345-23351.
• [11]Giacalone PL, Vuaroqueaux V, Daures JP, Houafic L, Martin PM, Laffargue F, Maudelonde T: Expression of adrenomedullin in human ovaries, ovarian cysts and cancers - Correlation with estrogens receptor status. Eur J Obstet Gynecol Reprod Biol 2003, 110(2):224-229.
• [12]Zhang Y, Zhang S, Shang H, Pang X, Zhao Y: Basic fibroblast growth factor upregulates adrenomedullin expression in ovarian epithelial carcinoma cells via JNK-AP-1 pathway. Regul Pept 2009, 157(1-3):44-50.
• [13]Springer TA, Wang JH: The three-dimensional structure of integrins and their ligands, and conformational regulation of cell adhesion. Cell Surface Receptors 2004, 68:29.
• [14]Buczek-Thomas JA, Chen N, Hasan T: Integrin-mediated adhesion and signalling in ovarian cancer cells. Cell Signal 1998, 10(1):55-63.
• [15]Reuning U: Integrin alpha v beta 3 Promotes Vitronectin Gene Expression in Human Ovarian Cancer Cells by Implicating Rel Transcription Factors. J Cell Biochem 2011, 112(7):1909-1919.
• [16]Sawada K, Mitra AK, Radjabi AR, Bhaskar V, Kistner EO, Tretiakova M, Jagadeeswaran S, Montag A, Becker A, Kenny HA, et al.: Loss of E-cadherin promotes ovarian cancer metastasis via alpha(5)-integrin, which is a therapeutic target. Cancer Res 2008, 68(7):2329-2339.
• [17]Mitra AK, Sawada K, Tiwari P, Mui K, Gwin K, Lengyel E: Ligand-independent activation of c-Met by fibronectin and alpha(5)beta(1)-integrin regulates ovarian cancer invasion and metastasis. Oncogene 2011, 30(13):1566-1576.
• [18]Morozevich G, Kozlova N, Cheglakov I, Ushakova N, Berman A: Integrin alpha 5 beta 1 controls invasion of human breast carcinoma cells by direct and indirect modulation of MMP-2 collagenase activity. Cell Cycle 2009, 8(14):2219-2225.
• [19]Ramakrishnan V, Bhaskar V, Law DA, Wong MHL, DuBridge RB, Breinberg D, O'Hara C, Powers DB, Liu G, Grove J, et al.: Preclinical evaluation of an anti-alpha5beta1 integrin antibody as a novel anti-angiogenic agent. J Exp Ther Oncol 2006, 5(4):273-286.
• [20]Ricart AD, Tolcher AW, Liu G, Holen K, Schwartz G, Albertini M, Weiss G, Yazji S, Ng C, Wilding G: Volociximab, a Chimeric Monoclonal Antibody that Specifically Binds alpha(5)beta(1) Integrin: A Phase I, Pharmacokinetic, and Biological Correlative Study. Clin Cancer Res 2008, 14(23):7924-7929.
• [21]Bell-McGuinn KM, Matthews CM, Ho SN, Barve M, Gilbert L, Penson RT, Lengyel E, Palaparthy R, Gilder K, Vassos A, et al.: A phase II, single-arm study of the anti-alpha 5 beta 1 integrin antibody volociximab as monotherapy in patients with platinum-resistant advanced epithelial ovarian or primary peritoneal cancer. Gynecol Oncol 2011, 121(2):273-279.
• [22]Bearz A, Tell G, Formisano S, Merluzzi S, Colombatti A, Pucillo C: Adhesion to fibronectin promotes the activation of the p125(FAK)/Zap-70 complex in human T cells. Immunology 1999, 98(4):564-568.
• [23]Shi Q, Boettiger D: A novel mode for integrin-mediated signaling: Tethering is required for phosphorylation of FAK Y397. Mol Biology Cell 2003, 14(10):4306-4315.
• [24]Tanaka T, Yamaguchi R, Sabe H, Sekiguchi K, Healy JM: Paxillin association in vitro with integrin cytoplasmic domain peptides. FEBS Lett 1996, 399(1-2):53-58.
• [25]Bellis SL, Miller JT, Turner CE: Characterization of tyrosine phosphorylation of paxillin in-vitro by focal adhesion kinase. J Biol Chem 1995, 270(29):17437-17441.
• [26]Schaller MD, Otey CA, Hildebrand JD, Parsons JT: Focal adhesion kinase and paxillin bind to peptides mimicking beta-integrin cytoplasmic domains. J Cell Biology 1995, 130(5):1181-1187.
• [27]Petit V, Boyer B, Lentz D, Turner CE, Thiery JP, Valles AM: Phosphorylation of tyrosine residues 31 and 118 on paxillin regulates cell migration through an association with CRK in NBT-II cells. J Cell Biology 2000, 148(5):957-969.
• [28]Tsubouchi A, Sakakura J, Yagi R, Mazaki Y, Schaefer E, Yano H, Sabe H: Localized suppression of RhoA activity by Tyr31/118-phosphorylated paxillin in cell adhesion and migration. J Cell Biology 2002, 159(4):673-683.
• [29]Kioon M-DA, Asensio C, Ea H-K, Uzan B, Cohen-Solal M, Liote F: Adrenomedullin increases fibroblast-like synoviocyte adhesion to extracellular matrix proteins by upregulating integrin activation. Arthritis Research & Therapy 2010, 12(5):R190. BioMed Central Full Text