Journal of Semnan University of Medical Sciences
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Outline
Expression and characterization of recombinant human epidermal growth factor receptor antigene (HER-2) as an indicator of breast cancer in yeast fermented systems
Authors
Abstract
Introduction: Immunotherapy has been practiced for more than two decades in the treatment of cancer. In this method, monoclonal antibodies bind to tumor specific antigens and inhibit their function. Therefore, scientists are looking for special targets on cancer cells that can be differentiate cancer cells from normal cells. In some cancers, including breast cancer human epidermal growth factor receptor-2 (HER-2) is significantly higher than the normal level. The goal for this project was to obtain expression with high quality of HER-2 with high activity in yeast expression system and its use in future studies. Materials and Methods: In order to express the extracellular domain of the HER-2 receptor we have used pPICZα and Pichia pastoris wild type X33 as a sequence vector and host cell respectively. Expression of HER-2 was analyzed using Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis 7% and accuracy of expression was confirmed by ELISA. Results: The expression of Her-2 antigen was evaluated by 7% SDS-PAGE with 185 KDa protein ladder. The results indicated that the expression of Her-2 antigen has been increased by IPTG induction. Moreover, the activity of Her-2 antigen was studied by ELISA technique using commercial anti Her-2 monoclonal antibody (Herceptin) and Her-2 antigen has shown high affinity to Herceptin. Conclusion: In this study, we have produced the Her-2 antigen in yeast expression system. With regards to yeast expression system advantages including secretory expression, protein folding, protein glycosylation, and activity of expressed protein in a wide range of pH, the Her-2 antigen which expressed in yeast host has great quality and will be used in future studies
Highlights
References
- 1.Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T, et al. Cancer statistics, 2008. Cancer J Clin 2008; 58: 71-96.
- 2.von Minckwitz G, Du Bois A, Schmidt M, Maass N, Cufer T, de Jongh FE, et al. Trastuzumab beyond progression in human epidermal growth factor receptor 2positive advanced breast cancer: A German Breast Group 26/Breast International Group 03-05 study. J Clin Oncol 2009; 27: 1999-2006.
- 3.Coussens L, Yang-Feng TL, Liao YC, Chen E, Gray A, McGrath J, et al. Tyrosine kinase receptor with extensive homology to EGF receptor shares chromosomal location with neu oncogene. Science 1985; 230: 1132-1139.
- 4.Ross JS, Fletcher JA, Bloom KJ, Linette GP, Stec J, Symmans WF, et al. Targeted therapy in breast cancer the HER-2/neu gene and protein. Mol Cell Proteomics 2004; 3: 379-398.
- 5.Brandt-Rauf PW, Pincus MR, Carney WP. The c-erbB-2 protein in oncogenesis: molecular structure to molecular epidemiology. CRO 1994; 5.
- 6.Ferguson KM, Darling PJ, Mohan MJ, Macatee TL, Lemmon MA. Extracellular domains drive homobut not heterodimerization of erbB receptors. EMBO 2000; 19: 4632-4643.
- 7.Mitsudomi T, Yatabe Y. Epidermal growth factor receptor in relation to tumor development: EGFR gene and cancer. FEBS 2010; 277: 301-308.
- 8.Olayioye MA, Neve RM, Lane HA, Hynes NE. The ErbB signaling network: receptor heterodimerization in development and cancer. EMBO 2000; 19: 3159-3167.
- 9.Yarden Y, Sliwkowski MX. Untangling the ErbB signalling network. Nat Rev Mol Cell Biol 2001; 2: 127-137.
- 10.Cho HS, Mason K, Ramyar KX, Stanley AM, Gabelli SB, Denney DW, et al. Structure of the extracellular region of HER2 alone and in complex with the Herceptin Fab. Nature 2003; 421: 756-760.
- 11.Tzahar E, Waterman H, Chen X, Levkowitz G, Karunagaran D, Lavi S, et al. A hierarchical network of interreceptor interactions determines signal transduction by Neu differentiation factor/neuregulin and epidermal growth factor. Mol Cell Biol 1996; 16: 5276-5287.
- 12.Mishani E, Hagooly A. Strategies for molecular imaging of epidermal growth factor receptor tyrosine kinase in cancer. J Nucl Med 2009; 50: 1199-1202.
- 13.Press MF, Slamon DJ, Flom KJ, Park J, Zhou J-Y, Bernstein L. Evaluation of HER-2/neu gene amplification and overexpression: comparison of frequently used assay methods in a molecularly characterized cohort of breast cancer specimens. J Clin Oncol 2002; 20: 3095-3105.
- 14.Romond EH, Perez EA, Bryant J, Suman VJ, Geyer Jr CE, Davidson NE, et al. Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer. N Engl J Med 2005; 353: 1673-1684.
- 15.Telli ML, Hunt SA, Carlson RW, Guardino AE. Trastuzumab-related cardiotoxicity: calling into question the concept of reversibility. J Clin Oncol 2007; 25: 3525-3533.
- 16.Ali SM, Carney WP, Esteva FJ, Fornier M, Harris L, Kstler W, et al. Serum HER2/neu and relative resistance to trastuzumabbased therapy in patients with metastatic breast cancer. Cancer 2008; 113: 1294-1301.
- 17.Cregg JM, Cereghino JL, Shi J, Higgins DR. Recombinant protein expression in Pichia pastoris. Mol Biotechnol 2000; 16: 23-52.
- 18.Yousefian S, Ranaei siadat O, Dehnavi E, Borjian Burojeni M, Nikzad jamnani F. Secretive expression of bacterial -xylosidase gene including hexahistidin-tag in Pichia pastoris. Koomesh 2013; 14: 389-395.
- 19.Akbari Eidgahi MR, Nasr R, Shaebani AA, Moghbeli M. Isolation and cloning of the subunit of human follicle stimulating Hormone (hFSH) with its native gene's signal sequence in methylotroph yeast Pichia pastoris pPIC9 shuttle vector. Koomesh 2006; 8: 11-18.
- 20.Cregg JM, Tolstorukov I, Kusari A, Sunga J, Madden K, Chappell T. Expression in the yeast Pichia pastoris. Methods Enzymol 2009; 463: 169-189.
- 21.Sheikholeslami F, Rasaee MJ, Shokrgozar MA, Dizaji MM, Rahbarizadeh F, Ahmadvande D. Isolation of a novel nanobody against HER-2/neu using phage displays technology. Lab Med 2010; 41: 69-76.
- 22.Sohi AN, Rajabibazl M, Rasaee M, Omidfar K. The use of camel antibodies in development of EGFRvIII enzyme-linked immunosorbent assay. Appl Biochem Microbiol 2015; 51: 374-380.
- 23.Kraus MH, Popescu N, Amsbaugh S, King CR. Overexpression of the EGF receptor-related proto-oncogene erbB-2 in human mammary tumor cell lines by different molecular mechanisms. EMBO 1987; 6: 605.
- 24.Corso S, Ghiso E, Cepero V, Sierra JR, Migliore C, Bertotti A, et al. Activation of HER family members in gastric carcinoma cells mediates resistance to MET inhibition. Mol Cancer 2010; 9.
- 25.Slamon DJ, Godolphin W, Jones LA, Holt JA, Wong SG, Keith DE, et al. Studies of the HER-2/neu proto-oncogene in human breast and ovarian cancer. Science 1989; 244: 707-712.
- 26.Tuefferd M, Couturier J, Penault-Llorca F, Vincent-Salomon A, Brot P, Guastalla J-P, et al. HER2 status in ovarian carcinomas: a multicenter GINECO study of 320 patients. PloS One 2007; 2: e1138.
- 27.Wilking U, Karlsson E, Skoog L, Hatschek T, Lidbrink E, Elmberger G, et al. HER2 status in a population-derived breast cancer cohort: discordances during tumor progression. Breast Cancer Res Treat 2011; 125: 553-561.
- 28.Boettner M, Prinz B, Holz C, Stahl U, Lang C. High-throughput screening for expression of heterologous proteins in the yeast Pichia pastoris. J Biotechnol 2002; 99: 51-62.
- 29.Vlahopoulos S, Gritzapis AD, Perez SA, Cacoullos N, Papamichail M, Baxevanis CN. Mannose addition by yeast Pichia Pastoris on recombinant HER-2 protein inhibits recognition by the monoclonal antibody herceptin. Vaccine 2009; 27: 4704-4708.##.
Copyright
© 2016, Author(s). This open-access article is available under the Creative Commons Attribution 4.0 (CC BY 4.0) International License (https://creativecommons.org/licenses/by/4.0/), which allows for unrestricted use, distribution, and reproduction in any medium, provided that the original work is properly cited.
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