Effect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy

Xun Huang,1,2 Jiexiang He,2,3 Huan-tian Zhang,1 Kai Sun,2,3 Jie Yang,1 Huajun Wang,1 Hongxin Zhang,2,3 Zhenzhao Guo,2,3 Zhen-gang Zha,1 Changren Zhou2,31Department of Bone and Joint Surgery, The First Affiliated Hospital of Jinan University, 2Department of Materials Science and Engineering, Jinan Un...

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Autores principales:	Huang X, He JX, Zhang HT, Sun K, Yang J, Wang HJ, Zhang HX, Guo ZZ, Zha ZG, Zhou CR
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2017
Materias:	AFM tumour dynamic force spectroscopy kinetic and thermodynamic interactions affinity nanoindentation Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/d65d859fd69d4285835d6cce323c8e9e
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spelling	oai:doaj.org-article:d65d859fd69d4285835d6cce323c8e9e2021-12-02T07:20:22ZEffect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy1178-2013https://doaj.org/article/d65d859fd69d4285835d6cce323c8e9e2017-12-01T00:00:00Zhttps://www.dovepress.com/effect-of-dacarbazine-on-cd44-in-live-melanoma-cells-as-measured-by-at-peer-reviewed-article-IJNhttps://doaj.org/toc/1178-2013Xun Huang,1,2 Jiexiang He,2,3 Huan-tian Zhang,1 Kai Sun,2,3 Jie Yang,1 Huajun Wang,1 Hongxin Zhang,2,3 Zhenzhao Guo,2,3 Zhen-gang Zha,1 Changren Zhou2,31Department of Bone and Joint Surgery, The First Affiliated Hospital of Jinan University, 2Department of Materials Science and Engineering, Jinan University, 3Engineering Research Center of Artificial Organs and Materials, Ministry of Education, Guangzhou, ChinaAbstract: CD44 ligand–receptor interactions are known to be involved in regulating cell migration and tumor cell metastasis. High expression levels of CD44 correlate with a poor prognosis of melanoma patients. In order to understand not only the mechanistic basis for dacarbazine (DTIC)-based melanoma treatment but also the reason for the poor prognosis of melanoma patients treated with DTIC, dynamic force spectroscopy was used to structurally map single native CD44-coupled receptors on the surface of melanoma cells. The effect of DTIC treatment was quantified by the dynamic binding strength and the ligand-binding free-energy landscape. The results demonstrated no obvious effect of DTIC on the unbinding force between CD44 ligand and its receptor, even when the CD44 nanodomains were reduced significantly. However, DTIC did perturb the kinetic and thermodynamic interactions of the CD44 ligand–receptor, with a resultant greater dissociation rate, lower affinity, lower binding free energy, and a narrower energy valley for the free-energy landscape. For cells treated with 25 and 75 µg/mL DTIC for 24 hours, the dissociation constant for CD44 increased 9- and 70-fold, respectively. The CD44 ligand binding free energy decreased from 9.94 for untreated cells to 8.65 and 7.39 kcal/mol for DTIC-treated cells, which indicated that the CD44 ligand–receptor complexes on DTIC-treated melanoma cells were less stable than on untreated cells. However, affinity remained in the micromolar range, rather than the millimolar range associated with nonaffinity ligands. Hence, the CD44 receptor could still be activated, resulting in intracellular signaling that could trigger a cellular response. These results demonstrate DTIC perturbs, but not completely inhibits, the binding of CD44 ligand to membrane receptors, suggesting a basis for the poor prognosis associated with DTIC treatment of melanoma. Overall, atomic force microscopy-based nanoscopic methods offer thermodynamic and kinetic insight into the effect of DTIC on the CD44 ligand-binding process.Keywords: atomic force microscopy, tumor, dynamic force spectroscopy, kinetic and thermodynamic interactions, affinity, nanoindentationHuang XHe JXZhang HTSun KYang JWang HJZhang HXGuo ZZZha ZGZhou CRDove Medical PressarticleAFMtumourdynamic force spectroscopykinetic and thermodynamic interactionsaffinitynanoindentationMedicine (General)R5-920ENInternational Journal of Nanomedicine, Vol Volume 12, Pp 8867-8886 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	AFM tumour dynamic force spectroscopy kinetic and thermodynamic interactions affinity nanoindentation Medicine (General) R5-920
spellingShingle	AFM tumour dynamic force spectroscopy kinetic and thermodynamic interactions affinity nanoindentation Medicine (General) R5-920 Huang X He JX Zhang HT Sun K Yang J Wang HJ Zhang HX Guo ZZ Zha ZG Zhou CR Effect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy
description	Xun Huang,1,2 Jiexiang He,2,3 Huan-tian Zhang,1 Kai Sun,2,3 Jie Yang,1 Huajun Wang,1 Hongxin Zhang,2,3 Zhenzhao Guo,2,3 Zhen-gang Zha,1 Changren Zhou2,31Department of Bone and Joint Surgery, The First Affiliated Hospital of Jinan University, 2Department of Materials Science and Engineering, Jinan University, 3Engineering Research Center of Artificial Organs and Materials, Ministry of Education, Guangzhou, ChinaAbstract: CD44 ligand–receptor interactions are known to be involved in regulating cell migration and tumor cell metastasis. High expression levels of CD44 correlate with a poor prognosis of melanoma patients. In order to understand not only the mechanistic basis for dacarbazine (DTIC)-based melanoma treatment but also the reason for the poor prognosis of melanoma patients treated with DTIC, dynamic force spectroscopy was used to structurally map single native CD44-coupled receptors on the surface of melanoma cells. The effect of DTIC treatment was quantified by the dynamic binding strength and the ligand-binding free-energy landscape. The results demonstrated no obvious effect of DTIC on the unbinding force between CD44 ligand and its receptor, even when the CD44 nanodomains were reduced significantly. However, DTIC did perturb the kinetic and thermodynamic interactions of the CD44 ligand–receptor, with a resultant greater dissociation rate, lower affinity, lower binding free energy, and a narrower energy valley for the free-energy landscape. For cells treated with 25 and 75 µg/mL DTIC for 24 hours, the dissociation constant for CD44 increased 9- and 70-fold, respectively. The CD44 ligand binding free energy decreased from 9.94 for untreated cells to 8.65 and 7.39 kcal/mol for DTIC-treated cells, which indicated that the CD44 ligand–receptor complexes on DTIC-treated melanoma cells were less stable than on untreated cells. However, affinity remained in the micromolar range, rather than the millimolar range associated with nonaffinity ligands. Hence, the CD44 receptor could still be activated, resulting in intracellular signaling that could trigger a cellular response. These results demonstrate DTIC perturbs, but not completely inhibits, the binding of CD44 ligand to membrane receptors, suggesting a basis for the poor prognosis associated with DTIC treatment of melanoma. Overall, atomic force microscopy-based nanoscopic methods offer thermodynamic and kinetic insight into the effect of DTIC on the CD44 ligand-binding process.Keywords: atomic force microscopy, tumor, dynamic force spectroscopy, kinetic and thermodynamic interactions, affinity, nanoindentation
format	article
author	Huang X He JX Zhang HT Sun K Yang J Wang HJ Zhang HX Guo ZZ Zha ZG Zhou CR
author_facet	Huang X He JX Zhang HT Sun K Yang J Wang HJ Zhang HX Guo ZZ Zha ZG Zhou CR
author_sort	Huang X
title	Effect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy
title_short	Effect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy
title_full	Effect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy
title_fullStr	Effect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy
title_full_unstemmed	Effect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy
title_sort	effect of dacarbazine on cd44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy
publisher	Dove Medical Press
publishDate	2017
url	https://doaj.org/article/d65d859fd69d4285835d6cce323c8e9e
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Effect of dacarbazine on CD44 in live melanoma cells as measured by atomic force microscopy-based nanoscopy

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