Abstract
Canine mammary gland tumor (MGT) is the commonest tumor in female dogs and a good animal model of human breast cancer. A group of newly identified genes encoding secreted frizzled-related proteins (SFRP) have been implicated in apoptosis regulation and tumorigenesis. Canine mammary tissues from 50 spontaneous MGTs and 10 normal mammary glands (MGs) were obtained from surgically excised specimens and analyzed for expression of SFRP2, β-catenin, and cyclin D1. By RT-PCR and in situ hybridization, SFRP2 gene was found abundantly expressed in neoplastic mammary tissues but not in normal mammary tissues, suggesting that SFRP2 may contribute as a tumor marker in canine MGTs. By immunohistochemical staining, the immunoreactivity of the SFRP2 protein was detected in more diverse areas than SFRP2 mRNA expression, including nuclei or/and cytoplasm and extracellular matrix of the tumor. In tumor masses, β-catenin lost its tight association with the membrane and diffused into the nucleus. The expression of β-catenin (79.4% positive) and cyclin D1 (71.4% positive) was also increased in MGTs. In the course of tumor progression, SFRP2 mRNA (p < 0.05) and β-catenin protein (p < 0.01) steadily increased but not in cyclin D1. The level of SFRP2 was linearly correlated with its downstream target β-catenin (p < 0.05), but not correlated with cyclin D1 (p < 0.5). As revealed in this study, the exclusive overexpression of SFRP2 in canine MGTs suggests that SFRP2 is a potential candidate gene for further investigation of mammary tumorigenesis and complex etiology of the canine model of mammary neoplasms.
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Nusse R, Varmus HE: Wnt genes. Cell 69: 1073–1087, 1992
Parr BA, McMahon AP: Wnt genes and vertebrate development. Genet Dev 4: 523–528, 1994
Moon RT, Brown JD, Torres M: WNTs modulate cell fate and behavior during vertebrate development. Trends Genet 13: 157–162, 1997
Gubb D, Garcia-Bellido A: A genetic analysis of the determination of cuticular polarity during development in Drosophila melanogaster. J Embryol Exp Morphol 68: 37–57, 1982
Bhanot P, Brink M, Samos CH, Hsieh JC, Wang Y, Macke JP, Andrew D, Nathans J, Nusse R: A new member of the frizzled family from Drosophila function as a Wingless receptor. Nature 382: 225–230, 1996
Finch PW, He X, Kelley MJ, Ñren A, Schaudies RP, Popescu NC, Rudikoff S, Aaronson SA, Varmus HE, Rubin JS: Puri-fication and molecular cloning of a secreted, frizzled-related antagonist of WNT action. Proc Natl Acad Sci USA 94: 6770–6775, 1997
Rattner A, Hsieh JC, Smallwood PM, Gilbert DJ, Copeland NG, Jenkins NA, Nathans J: A family of secreted proteins contains homology to the cysteine-rich ligand-binding domain of frizzled receptors. Proc Natl Acad Sci USA 94: 2859–2863, 1997
Melkonyan HS, Chang WC, Shapiro JP, Mahedevappa M, Fitzpatrick PA, Kiefer MC, Tomei LD, Urmansky SR: sARPs: a family of secreted apoptosis-related proteins. Proc Natl Acad Sci USA 94: 13636–13641, 1997
Wolf V, Ke G, Dharmarajan AM, Bielke W, Artuso L, Saurer S, Friis R: DDC-4, an apoptosis-associated gene, is a secreted frizzled relative. FEBS Lett 417: 385–389, 1997
Lin CT, Sargan DR: A method for generating subtractive cDNA libraries retaining clones containing repetitive elements. Nucl Acids Res 25: 4427–4428, 1997
Lin CT, Sargan DR: Generation and analysis of canine retinal ESTs: isolation and expression of retina-specific gene transcripts. Biochem Biophys Res Commun 282: 394–403, 2001
Lammers CH, D'Souza U, Qin ZH, Lee SH, Yajima S, Mouradian MM: Regulation of striatal dopamine receptors by estrogen. Synapse 34: 222–227, 1999
Cardone A, Angelini F, Varriale B: Autoregulation of estrogen and androgen receptor mRNAs and downregulation of androgen receptor mRNA by estrogen in primary cultures of lizard testis cells. Gen Comp Endocrinol 110: 227–236, 1998
Nickenig G, Baumer AT, Grohe C, Kahlert S, Strehlow K, Rosenkranz S, Stablein A, Beckers F, Smits JF, Daemen MJ, Vetter H, Bohm M: Estrogen modulates AT1 receptor gene expression in vitro and in vivo. Circulation 97: 2197–2201, 1998
Batra S, Al-Hijji J: Characterization of nitric oxide synthase activity in rabbit uterus and vagina: downregulation by estrogen. Life Sci 62: 2093–2100, 1998
Jones SE, Jomary C, Grist J, Stewart HJ, Neal MJ: Altered expression of secreted frizzled-related protein-2 in retinitis pigmentosa retinas. Invest Ophthalmol Vis Sci 41: 1297–1301, 2000
Roth W, Wild-Bode C, Platten M, Grimmel C, Melkonyan HS, Dichgans J, Weller M: Secreted frizzled-related proteins inhibit motility and promote growth of human malignant glioma cells. Oncogene 19: 4210–4220, 2000
Nerurkar VR, Chitale BV, Jalnapurkar BV, Naik SN, Lalitha VS: Comparative pathology of canine mammary tumors. J Comp Pathol 101: 389–397, 1989
Destexhe E, Lespagnard L, Degeyter M, Heymann R, Coignoul F: Immunohistochemical identification of myoepithelial, epithelial, and connective tissue cells in canine mammary tumors. Vet Pathol 30: 146–154, 1993
Donnay I, Rauis J, Devleeschouwer N, Wouters-Ballman P, Leclercq G, Verstegen J: Comparison of estrogen and progesterone receptor expression in normal and tumor mammary tissues from dogs. Am J Vet Res 56: 1188–1194, 1995
Middleton LP, Chen V, Perkins GH, Pinn V, Page D: Histopathology of breast cancer among African American women. Cancer 97(Suppl 1): 253–257, 2003
Martin PM, Cotard M, Mialot JP, Andre F, Raynaud JP: Animal models for hormone-dependent human breast cancer. Relationship between steroid receptor profiles in canine and feline mammary tumors and survival rate. Cancer Chemother Pharmacol 12: 13–17, 1984
Donnay I, Rauis J, Wouters-Ballman P, Devleeschouwer N, Leclercq G., Verstegen JP: Receptors for oestrogen, progesterone and epidermal growth factor in normal and tumorous canine mammary tissues. J Reprod Fertil 47: 501–512, 1993
Lee JL, Chang CJ, Chueh LL, Lin CT: Expression of secreted frizzled related protein 2 in a primary canine mammary tumor SFRP2 is highly expressed in canine MGTs 149 cell line: a candidate tumor marker for mammary tumor cells. In Vitro Cell Dev Biol-Anim 39: 357-364
Remmele W, Schicketanz KH: Immunohistochemical determination of estrogen and progesterone receptor content in human breast cancer. Computer-assisted image analysis (QIC score) vs. subjective grading IRS. Pathol Res Pract 189: 862–866, 1993
Lin WC, Shen BJ, Tsay YG, Yen HC, Lee SC, Chang CJ: Transcriptional activation of C/EBPbeta gene by c-Jun and ATF2. DNA Cell Biol 21: 551–560, 2002
Chang CJ, Chen YL, Lee SC: Coactivator TIF1beta interacts with transcription factor C/EBPbeta and glucocorticoid receptor to induce alpha1-acid glycoprotein gene expression. Mol Cell Biol 18: 5880–5887, 1998
Sambrook J, Gritsch E, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, New York, 1989
Bergstein I, Brown AMC: In: Bow-cock AM (ed) Breast Cancer: Molecular Genetics, Pathogenesis and Therapeutics. Human Press, Totowa, NJ, 1999, pp 181–198
Morin PJ, Sparks AB, Korinek V, Barker N, Clevers H, Vogelstein B, Kinzler KW: Activation of â-catenin-Tcf signaling in colon cancer by mutations in α-catenin or APC. Science 275: 1787–1790, 1997
He TC, Sparks AB, Rago C, Hermeking H, Zawel L, da Costa LT, Morin PJ, Vogelstein B, Kinzler KW: Identification of c-MYC as a target of the APC pathway. Science 281: 1509–1512, 1998
Tetsu O, McCormick F: α-Catenin regulates expression of cyclin D1 in colon carcinoma cells. Nature 398: 422–426, 1999
Roose J, Huls G, van Beest M, Moerer P, van der Horn K, Goldschmeding R, Logtenberg T, Clevers H: Synergy between tumor suppressor APC and the â-catenin-Tcf4 target Tcf1. Science 285: 1923–1926, 1999
Ingvarsson S: Molecular genetics of breast cancer progression. Semin Cancer Bio 9: 277–288, 1999
Ross JS, Fletcher JA: HER-2/neu (c-erb-B2) gene and protein in breast cancer. Am J Clin Pathol 112: S53–S67, 1999
Simpson JF, Page DL: The p53 tumor suppressor gene in ductal carcinoma in situ of the breast. Am J Pathol 156: 5–6, 2000
Lin TP, Guzman RC, Osborn RC, Thordarson G, Nandi S: Role of endocrine, autocrine, and paracrine interactions in the development of mammary hyperplasia inWnt-1 transgenic mice. Cancer Res 52: 4413–4419, 1992
Chen S, Guttridge DC, You Z, Zhang Z, Fribley A, Mayo MW, Kitajewski J, Wang CY: Wnt-1 signaling inhibits apoptosis by activating â-catenin/T cell factor-mediated transcription. J Biol Chem 152: 87–96, 2001
Wang TC, Cardiff RD, Zukerburg L, Lees E, Arnold A, Schmidt EV: Mammary hyperplasia and carcinoma in MMTV-cyclin D1 transgenic mice. Nature 369: 669–671, 1994
Bodrug SE, Warner BJ, Bath ML, Lindeman GJ, Harris AW, Adams JM: Cyclin D1 transgene impedes lymphocyte maturation and collaborates in lymphomagenesis with the myc gene. EMBO J 13: 2124–2130, 1994
Bafico A, Gazit A, Pramila T, Finch PW, Yaniv A, Aaronson SA: Interaction of frizzled related protein (FRP) with wnt ligands and the frizzled receptor suggests alternative mechanisms for FRP inhibition of wnt signaling. J Biol Chem 274: 16180–16187, 1999
Xu W, D'Amore PA, Sokol SY: Functional and biochemical interactions of Wnts with FrzA, a secreted Wnt antagonist. Development 125: 4767–4776, 1998
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Lee, JL., Chang, CJ., Wu, SY. et al. Secreted Frizzled-related Protein 2 (SFRP2) is Highly Expressed in Canine Mammary Gland Tumors but not in Normal Mammary Glands. Breast Cancer Res Treat 84, 139–149 (2004). https://doi.org/10.1023/B:BREA.0000018412.83348.ff
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DOI: https://doi.org/10.1023/B:BREA.0000018412.83348.ff