Abstract
The ERBB2 oncogene encodes a trans-membrane glycoprotein (p185) with tyrosine kinase activity, which is closely related to epidermal growth factor receptor (EGFR). All four members of the family have been reported to be expressed in breast cancer cells in vitro. ERBB2 expression predicts poor prognosis in cancer. The present study has evaluated the association of the expression levels of ERBB2 with mammary tumor development in dogs and checked the heterologous expression of ERBB2 protein in prokaryotic system. Gene expression was measured using qRT-PCR. Mammary tissue samples were collected from healthy canine mammary gland (n = 10) and from tumor subjects (n = 10). ERBB2 expression was found to be upregulated (5.659 times) in canine mammary tumor state suggesting malignant behavior of canine mammary tumor association with increased transcription of ERBB2 gene. Ectodomain encoding ERBB2 gene was cloned in E. coli and the recombinant protein produced was successfully purified by Ni–NTA affinity chromatography which was further confirmed by western blotting. Recombinant ERBB2 protein can be used as a marker for prognosis of canine mammary tumor.
Similar content being viewed by others
References
Akiyama T, Sudo C, Ogawara H, Toyoshima K, Yamamoto T (1986) The product of the human c-ERBB2 gene: a 185-kilodalton glycoprotein with tyrosine kinase activity. Science 232:1644–1646
Plowman GD, Whitney GS, Neubauer MG (1990) Molecular cloning and expression of an additional epidermal growth factor receptor-related gene. Proc Natl Acad Sci 87:4905–4909
Plowman GD, Culouscou JM (1993) Whitney GS (1993) Ligand specific activation of HER4/p180erB4, a fourth member of the epidermal growth factor receptor family. Proc Natl Acad Sci 90:1746–1750
Lupu R, Cardillo M, Harris L, Hijazi M, Rosenberg K (1995) Interaction between ERBB-receptors and heregulin in breast cancer tumor progression and drug resistance. Semin Cancer Biol 6:135–145
Gschwind A, Fischer OM, Ullrich A (2004) The discovery of receptor tyrosine kinases: targets for cancer therapy. Nat Rev Cancer 4:361–370
Schlessinger J (2004) Common and distinct elements in cellular signaling via EGF and FGF receptors. Science 306:1506–1507
Bertucci F, Borie N, Ginestier C, Groulet A, Charafe-Jauffret E, Adélaïde J, Geneix J, Bachelart L, Finetti P, Koki A, Hermitte F, Hassoun J, Debono S, Viens P, Fert V, Jacquemier J, Birnbaum D (2004) Identification and validation of an ERBB2 gene expression signature in breast cancers. Oncogene 23(14):2564–2575
Landgraf R (2007) HER2 therapy. HER2 (ERBB2): functional diversity from structurally conserved building blocks. Breast Cancer Res 9(1):202
Bernstein JL, Godbold JH, Raptis G, Watson MA, Levinson B, Aaronson SA, Fleming TP (2005) Identification of mammaglobin as a novel serum marker for breast cancer. Clin Cancer Res 11:6528–6535
Benjamin S, Lee A, Saunders W (1999) Classification and behavior of canine mammary epithelial neoplasms based on life-span observations in beagles. Vet Pathol 36(5):423–436
Ettinger SJ, Feldman EC (1995) Textbook of veterinary internal medicine, 4th edn. W.B. Saunders Company, Philadelphia
Klopfleisch R, Lenze D, Hummel M, Gruber AD (2011) The metastatic cascade is reflected in the transcriptome of metastatic canine mammary carcinomas. Vet J 190:236–243
Klopfleisch R, Klose P, Weise C, Bondzio A, Multhaup G, Einspanier R, Gruber AD (2010) The proteome of metastatic canine mammary carcinomas: similarities and differences to human breast cancer. J Proteome Res 9(12):6380–6391
Karayannopoulou M, Kaldrymidou E, Constantinidis TC, Dessiris A (2005) Histological grading and prognosis in dogs with mammary carcinomas: application of a human grading method. J Comp Pathol 133:246–252
Chomczynski P, Sacchi N (1987) Single step method of RNA isolation by guanidium thiocyanate–phenol-chloroform extraction. Anal Biochem 162:156–159
Peruzzi D, Mori F, Conforti F, Lazzaro D, De Rinaldis E, Nicola La Monica G, Aurisicchio L (2009) MMP-11: a novel target antigen for cancer immunotherapy. Clin Cancer Res 15:4104–4109
Abasht B, Kaiser MG, Van der Poel J, Lamont SJ (2009) Genetic lines differ in Toll-like receptor gene expression in spleen of chicks inoculated with Salmonella enterica serovar Enteritidis. Poult Sci 88:744–749
Livak KJ, Schmittgen TD (2001) Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2-[Delta][Delta]CT Method. Methods 25(4):402–408
Sambrook J, Russel DW (2001) Molecular cloning: a laboratory manual, 3rd edn. Cold spring harbor laboratory press, New York
Towbin H, Staehelin T, Gordin J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Siczak-Kuta A, Tomaszewska R, Rudnicka-Sosin L, Oko K, Stachura J (2007) Evaluation of HER2/neu gene amplification in patients with invasivebreast carcinoma. Comparison of in situ hybridization methods. Pol J Pathol 58(1):41–50
Slamon DJ, Clark GM, Wong SG, Levin WJ, Ullrich A, McGuire WL (1987) Human breast cancer: correlation of relapse and survival with amplification of the HER-2/neu oncogene. Science 235(4785):177–182
Zhou Y, Zhang J, Jin H, Chen Z, Wu Q, Li W, Yue M, Luo C, Wang M (2013) Prokaryotic expression and refolding of EGFR extracellular domain and generation of phage display human scFv against EGFR. Biomed Pharmacother 67:737–743
O’Connor S, Li E, Majors BS, He L, Placone J, Baycin D, Betenbaugh MJ, Hristova K (2009) Increased expression of the integral membrane protein ErbB2 in Chinese hamster ovary cells expressing the anti-apoptotic gene Bcl-xL. Prot Exp Pur 67(1):41–47
Biburger M, Weth R, Wels WS (2005) A Novel Bispecific Tetravalent Antibody Fusion Protein to Target Costimulatory Activity for T-cell Activation to Tumor Cells Overexpressing ErbB2/HER2. J Mol Biol 346:1299–1311
Acknowledgments
The authors are thankful to the Department of Biotechnology, India, for providing funds to carry out the research work. None of the authors of this paper has a financial or personal relationship with other people or organisations that could inappropriately influence or bias the content of the paper.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Chaudhary, N., Sunil Kumar, B.V., Bhardwaj, R. et al. Expression of Erythroblastic Leukemia Viral Oncogene Homolog 2 (ERBB2) from Canine Mammary Tumor. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 85, 801–806 (2015). https://doi.org/10.1007/s40011-014-0455-z
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s40011-014-0455-z