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. 2020 Nov 19;21(22):8760. doi: 10.3390/ijms21228760

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Propranolol influences adaptation of cells to hypoxic microenvironment and decreases level of CA IX. (A) Immunohistochemical analysis of CA IX and Ki-67 expression and localization in HCT116 spheroids sections. CA IX was expressed in the hypoxic centre and perinecrotic regions, a strong expression was observed in the regions very close to the spheroid surface. Expression of CA IX was detected in areas of proliferative Ki-67 positive cells. Localization and distribution of CA IX in spheroids is, independently of propranolol treatment, polarized and non-uniform. Proliferative (Ki-67 positive) and CA IX positive cells are localized in the field opposite to necrotic cells area. (B) Immunofluorescent analysis of CA IX expression in HCT116 and HT29 spheroids. Reduced expression of CA IX was confirmed also by immunofluorescence. The central regions of the HCT116 and HT29 spheroid were necrotic after 5 days of β-AR inhibition, while the integrity of the control spheroids remained intact.