Effect of hyaluronic acid on the drug resistance of 3D in vitro models of brain tumors



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BACKGROUND: One of the key problems in the treatment of brain tumors is the development of drug resistance that appears during treatment, which significantly reduces the efficacy of standard chemotherapy methods. Three-dimensional cell models are a powerful tool in experimental oncology, allowing us to study the mechanisms of drug resistance in tumor cells, including the contribution of signaling pathways activated by extracellular matrix molecules.

AIM: To evaluate the effect of hyaluronic acid on the resistance of 3D in vitro brain tumor models to doxorubicin.

METHODS: 2D monolayer cultures were obtained from continuous brain tumor strains from laboratory animals stored in the unique collection of the A.P. Avtsyn Research Institute of Human Morphology, B.V. Petrovsky Russian Scientific Center of Surgery. 3D spheroids were formed under ultra-low adhesion conditions with or without hyaluronic acid. Doxorubicin cytotoxicity was determined using the CCK-8 assay. Doxorubicin efflux efficiency and CD44 protein production were determined using fluorimetry, and the proportion of CD44+ cells was determined using flow cytometry. The expression of genes associated with drug resistance was assessed using real-time PCR.

RESULTS: The IC50 values of doxorubicin were 180 nM for the 2D model of oligodendroglioma 51/7, 280 nM for glioblastoma 14-60-4, 500 nM for astrocytoma 10-17-2, and 2750 nM for neurinoma NGUK2. Doxorubicin at the IC50 concentration determined for the 2D models did not significantly affect the viability of cells within the spheroids, but a fourfold increase in its concentration resulted in a cytotoxic effect, with its severity depending on the degree of spheroid compaction. Hyaluronic acid dose-dependently increased the resistance of 3D brain tumor models to doxorubicin due to the activation of the CD44 receptor, more efficient drug efflux from cells, provided by changes in the expression of ABC transporter genes (Abcb1, Abcg2, Abcc1), as well as genes associated with DNA repair (Mgmt, Top2a).

CONCLUSION: The obtained data expand the understanding of the role of hyaluronic acid and the CD44 receptor in the formation of drug resistance in brain tumors, which allows for the creation of more relevant in vitro models. The results of the study are of practical importance for the development of strategies to overcome drug resistance, including targeted inhibition of ABC transporters or blockade of the interaction of hyaluronic acid and its receptor CD44.

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作者简介

Irina Arutyunyan

Avtsyn Research Institute of Human Morphology, Petrovsky National Research Centre of Surgery, Moscow, Russia; Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples’ Friendship University of Russia, Moscow, Russia

Email: labrosta@yandex.ru
ORCID iD: 0000-0002-4344-8943

MD, Cand. Sci. (Biology), Senior Researcher, Laboratory of Growth and Development

俄罗斯联邦, Moscow, Russia; Moscow, Russia

Georgy Lositsky

Avtsyn Research Institute of Human Morphology, Petrovsky National Research Centre of Surgery, Moscow, Russia; Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples’ Friendship University of Russia, Moscow, Russia

Email: glosierror404@gmail.com
俄罗斯联邦, Moscow, Russia; Moscow, Russia

Anna Soboleva

Avtsyn Research Institute of Human Morphology, Petrovsky National Research Centre of Surgery, Moscow, Russia; Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples’ Friendship University of Russia, Moscow, Russia

Email: annasobo@mail.ru
俄罗斯联邦, Moscow, Russia; Moscow, Russia

Sofia Aleksandrova

Avtsyn Research Institute of Human Morphology, Petrovsky National Research Centre of Surgery, Moscow, Russia; Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples’ Friendship University of Russia, Moscow, Russia

Email: sofia.aleksa@mail.ru
俄罗斯联邦, Moscow, Russia; Moscow, Russia

Dorzhu Balchir

Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples’ Friendship University of Russia, Moscow, Russia

Email: dbalchir@mail.ru
俄罗斯联邦, Moscow, Russia

Vera Kudelkina

Avtsyn Research Institute of Human Morphology, Petrovsky National Research Centre of Surgery, Moscow, Russia

Email: verakudelkina8047@gmail.com
俄罗斯联邦, Moscow, Russia

Andrey Makarov

Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples’ Friendship University of Russia, Moscow, Russia

Email: anvitmak@yandex.ru
俄罗斯联邦, Moscow, Russia

Andrey Elchaninov

Avtsyn Research Institute of Human Morphology, Petrovsky National Research Centre of Surgery, Moscow, Russia; Research Institute of Molecular and Cellular Medicine, Patrice Lumumba Peoples’ Friendship University of Russia, Moscow, Russia; National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov, Moscow, Russia

编辑信件的主要联系方式.
Email: elchandrey@yandex.ru
俄罗斯联邦, Moscow, Russia; Moscow, Russia; Moscow, Russia

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