Nanobiotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran; Endocrine Research Center, Institute of Endocrinology and Metabolism, Iran University of Medical Sciences, Tehran, Iran.
Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, Iran.
Int J Biol Macromol. 2022 Oct 31;219:1122-1134. doi: 10.1016/j.ijbiomac.2022.08.150. Epub 2022 Aug 28.
High expression of receptor tyrosine-protein kinase erbB-3 (HER3) has been found in several malignancies such as breast cancer. In this study, we designed, produced and evaluated a new affitoxin consisting of a truncated form of diphtheria toxin and a HER3-binding affibody domains. The new affitoxin was expressed in Escherichia coli and purified by affinity chromatography. We evaluated the suitability of affitoxin to kill HER3 positive breast cancer cells with MTT and apoptosis assays. The protein synthesis inhibition was also evaluated. The IC value in HER3 negative cells is about 10 times more than HER3 positive cells in new design of affitoxin. The specificity of affitoxin for binding to HER3 positive cells was also investigated with binding assay with flow cytometry. The results show that, the new affitoxin is an anti-cancer molecule with specific binding to HER3 positive cells and may open a new window for the treatment of HER3-positive cancers.
受体酪氨酸蛋白激酶 erbB-3（HER3）的高表达已在多种恶性肿瘤中被发现，如乳腺癌。在这项研究中，我们设计、生产并评估了一种由白喉毒素的截断形式和 HER3 结合 affibody 结构域组成的新型亲和毒素。新的亲和毒素在大肠杆菌中表达，并通过亲和层析进行纯化。我们通过 MTT 和凋亡测定评估了亲和毒素杀死 HER3 阳性乳腺癌细胞的适用性。还评估了蛋白合成抑制。在新型设计的亲和毒素中，HER3 阴性细胞的 IC 值比 HER3 阳性细胞高约 10 倍。我们还通过流式细胞术的结合测定研究了亲和毒素与 HER3 阳性细胞结合的特异性。结果表明，新型亲和毒素是一种针对 HER3 阳性细胞的抗癌分子，可能为治疗 HER3 阳性癌症开辟新的途径。
