Antiproliferative and apoptotic effects of anti-human HB-EGF neutralizing polyclonal antibodies in vitro
DOI:
https://doi.org/10.32471/exp-oncology.2312-8852.vol-42-no-1.14145Keywords:
antibodies., cancer treatment., HB-EGF, mitogenic activation, receptorsAbstract
Summary. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the epidermal growth factor family and has a variety of physiological and pathophysiological functions. Also, HB-EGF plays a pivotal role in progression of different tumors. So, HB-EGF seems to be a target molecule for the treatment of some cancer types. Aim: To obtain HB-EGF neutralizing polyclonal antibodies and test their anti-proliferative properties in vitro. Materials and Methods: Lab rabbits and mice were used for immunization with recombinant HB-EGF. The effect of generated polyclonal antibodies on viability and apoptosis of human epidermoid carcinoma derived A431 cell line was assessed using MTT and Annexin V-propidium iodide assays. Results: Rabbit polyclonal anti-HB-EGF serum could block binding of soluble HB-EGF to epidermal growth factor receptor/human epidermal growth factor receptor. Also, anti-HB-EGF antibodies could bind to surface of A431 cells which express abnormally high levels of membrane bound proHB-EGF and its receptor. It has been shown that immune serum with polyclonal antibodies against HB-EGF was able to block the mitogenic activation of the cells with HB-EGF and cause apoptotic cell death. Conclusion: Inhibition of HB-EGF activity with neutralizing polyclonal antibodies can effectively inhibit mitogenic activation and cause apoptosis of cancer cells with significant epidermal growth factor receptor overexpression.
References
Higashiyama S, Abraham JA, Miller J, et al. A heparin-binding growth factor secreted by macrophage-like cells that is related to EGF. Science 1991; 251: 936–9.
Higashiyama S, Lau K, Besner GE, et al. Structure of heparin-binding EGF-like growth factor. Multiple forms, primary structure, and glycosylation of the mature protein. J Biol Chem 1992; 267: 6205–12.
Roskoski R. The ErbB/HER family of protein-tyrosine kinases and cancer. Pharmacol Res 2014; 79: 34–74.
Prince RN, Schreiter ER, Zou P, et al. The heparin-binding domain of HB-EGF mediates localization to sites of cell-cell contact and prevents HB-EGF proteolytic release. J Cell Sci 2010; 123: 2308–18.
Mitamura T, Higashiyama S, Taniguchi N, et al. Diphtheria toxin binds to the epidermal growth factor (EGF)-like domain of human heparin-binding EGF-like growth factor/diphtheria toxin receptor and inhibits specifically its mitogenic activity. J Biol Chem 1995; 270: 1015–9.
Dao DT, Anez-Bustillos L, Adam RM, et al. Heparin-binding epidermal growth factor-like growth factor as a critical mediator of tissue repair and regeneration. Am J Pathol 2018; 188: 2446–56.
Iwamoto R, Yamazaki S, Asakura M, et al. Heparin-binding EGF-like growth factor and ErbB signaling is essential for heart function. Proc Natl Acad Sci U S A 2003; 100: 3221–6.
Jessmon P, Leach RE, Armant DR. Diverse functions of HBEGF during pregnancy. Mol Reprod Dev 2009; 76: 1116–27.
Ota I, Higashiyama S, Masui T, et al. Heparin-binding EGF-like growth factor enhances the activity of invasion and metastasis in thyroid cancer cells. Oncol Rep 2013; 30: 1593–600.
Yotsumoto F, Yagi H, Suzuki SO, et al. Validation of HB-EGF and amphiregulin as targets for human cancer therapy. Biochem Biophys Res Commun 2008; 365: 555–61.
Yotsumoto F, Sanui A, Fukami T, et al. Efficacy of ligand-based targeting for the EGF system in cancer. Anticancer Res 2009; 29: 4879–85.
Shen Y, Ruan L, Lian C, et al. Discovery of HB-EGF binding peptides and their functional characterization in ovarian cancer cell lines. Cell Death Discov 2019; 5: 82.
Nam SO, Yotsumoto F, Miyata K, et al. Anti-tumor effect of intravenous administration of CRM197 for triple-negative breast cancer therapy. Anticancer Res 2016; 36: 3651–7.
Tsujioka H, Fukami T, Yotsumoto F, et al. A possible clinical adaptation of CRM197 in combination with conventional chemotherapeutic agents for ovarian cancer. Anticancer Res 2011; 31: 2461–5.
Yagi H, Yotsumoto F, Sonoda K, et al. Synergistic anti-tumor effect of paclitaxel with CRM197, an inhibitor of HB-EGF, in ovarian cancer. Int J Cancer 2009; 124: 1429–39.
Dateoka S, Ohnishi Y, Kakudo K. Effects of CRM197, a specific inhibitor of HB-EGF, in oral cancer. Med Mol Morphol 2012; 45: 91–7.
Miyamoto S, Iwamoto R, Furuya A, et al. A novel anti-human HB-EGF monoclonal antibody with multiple antitumor mechanisms against ovarian cancer cells. Clin Cancer Res 2011; 17: 6733–41.
Sato S, Drake AW, Tsuji I, et al. A potent anti-HB-EGF monoclonal antibody inhibits cancer cell proliferation and multiple angiogenic activities of HB-EGF. PLoS One 2012; 7: e51964.
Hamaoka M, Chinen I, Murata T, et al. Anti-human HB-EGF monoclonal antibodies inhibiting ectodomain shedding of HB-EGF and diphtheria toxin binding. J Biochem 2010; 148: 55–69.
Iwamoto R, Takagi M, Akatsuka J, et al. Characterization of a novel anti-human HB-EGF monoclonal antibody applicable for paraffin-embedded tissues and diagnosis of HB-EGF-related cancers. Monoclon Antib Immunodiagn Immunother 2016; 35: 73–82.
Tsuji I, Sato S, Otake K, et al. Characterization of a variety of neutralizing anti-heparin-binding epidermal growth factor-like growth factor monoclonal antibodies by different immunization methods. MAbs 2012; 4: 732–9.
Sato S, Kamada H, Watanabe T, et al. Identification of the cancer cell proliferation and survival functions of proHB-EGF by using an anti-HB-EGF antibody. PLoS One 2013; 8: e54509.
Lipman NS, Jackson LR, Trudel LJ, et al. Monoclonal versus polyclonal antibodies: distinguishing characteristics, applications, and information resources. ILAR J 2005; 46: 258–68.
Korotkevych NV, Kolibo DV, Labyntsev AJ, et al. Obtaining of recombinant human heparin binding EGF-like growth factor and perspectives of its application in biotechnology. Biotekhnologia 2010; 3: 44–54.
Korotkevych NV, Labyntsev AJ, Kolybo DV, et al. Obtaining and characterization of recombinant fluorescent derivatives of soluble human HB-EGF. Biotechnol Acta 2014; 7: 46–53.
Schindelin J, Arganda-Carreras I, Frise E, et al. Fiji: an open-source platform for biological-image analysis. Nat Methods 2012; 9: 676–82.
Zhang Q, Chen G, Liu X, et al. Monoclonal antibodies as therapeutic agents in oncology and antibody gene therapy. Cell Res 2007; 17: 89–99.
Miyamoto S, Yagi H, Yotsumoto F, et al. Heparin-binding epidermal growth factor-like growth factor as a new target molecule for cancer therapy. Adv Exp Med Biol 2008; 622: 281–95.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2023 Experimental Oncology
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
