1. Antiproliferative and apoptotic effects of anti-human HB-EGF neutralizing polyclonal antibodies in vitro
- Author
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D. V. Kolybo, S. V. Komisarenko, O. I. Krynina, and A. A. Siromolot
- Subjects
Male ,Cancer Research ,medicine.medical_treatment ,Apoptosis ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Epidermal growth factor ,Cell Line, Tumor ,medicine ,Escherichia coli ,Animals ,Humans ,Epidermal growth factor receptor ,Cell Proliferation ,Mice, Inbred BALB C ,biology ,Chemistry ,Growth factor ,Immune Sera ,Antibodies, Monoclonal ,Molecular biology ,Antibodies, Neutralizing ,Recombinant Proteins ,Oncology ,Epidermoid carcinoma ,Cell culture ,Polyclonal antibodies ,030220 oncology & carcinogenesis ,biology.protein ,Female ,Immunization ,Rabbits ,Antibody ,A431 cells ,hormones, hormone substitutes, and hormone antagonists ,Heparin-binding EGF-like Growth Factor - Abstract
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.
- Published
- 2020