In vitro effects of ascorbic acid on viability and metabolism of patients’ osteosarcoma stem cells

• Published in: Acta Pharmaceutica Vol. 72; no. 4; pp. 599 - 613
• Main Authors: Jovičić, Marijana Šimić, Pušić, Maja, Antunović, Maja, Ledinski, Maja, Librenjak, Lucija, Kolundžić, Robert, Ribičić, Tomislav, Trkulja, Vladimir, Urlić, Inga
• Format: Journal Article Web Resource
• Language: English
• Published: Poland Sciendo 01.12.2022; De Gruyter Poland; Hrvatsko farmaceutsko društvo
• Subjects: Antineoplastic Agents - pharmacology; Antineoplastic Agents - therapeutic use; Antitumor agents; Ascorbic acid; Biopsy; Bone cancer; cancer stem cells; Cell culture; Cell Line, Tumor; Cell Proliferation; Cytotoxicity; Drug resistance; Glycolysis; HEK293 Cells; Humans; Incubation; Mesenchyme; Metabolism; Neoplastic Stem Cells - metabolism; Neoplastic Stem Cells - pathology; Osteosarcoma; Osteosarcoma - drug therapy; Osteosarcoma - metabolism; Osteosarcoma - pathology; Oxidative phosphorylation; Patients; Phosphorylation; Sarcoma; Stem cells; Toxicity; tumor cell metabolism; ascorbic acid; cancer stem cells; tumor cell metabolism; osteosarcoma
• ISSN: 1846-9558; 1330-0075; 1846-9558
• DOI: 10.2478/acph-2022-0040

Stagnation in novelties of osteosarcoma (OS) treatment indicates the need for new therapeutic methods. OS cancer stem cells (OS-CSC) are taught to have the ability to self-renew and develop mechanisms of anticancer drug resistance, and this is why it is difficult to eradicate them. Their metabolism has been recognized as a potential target of therapeutic action. Ascorbic acid (AA) is considered to act pro-oxidative against OS-CSC by oxidative effect and by inhibition of glycolysis. This study examined an impact of AA on OS-CSC metabolism isolated from patients’ biopsies, with the aim of better understanding of OS-CSC metabolism and the action of AA on OS-CSC. OS-CSC were isolated using a sphere culture system and identified as stem cells using Hoechst 33342 exclusion assay. Determination of the dominant type of metabolism of OS-CSC, parental OS cells, human mesenchymal stem cells (hMSC) and U2OS OS lineage before and after AA treatment was done by Seahorse XF (Agilent). Cytotoxicity of high-dose AA was confirmed by the MTT test and was proven for all the examined cell types as well as HEK293. Seahorse technology showed that OS-CSC can potentially use both glycolysis and oxidative phosphorylation (OXPHOS), and can turn to glycolysis and slow metabolic potential in unfavorable conditions such as incubation in AA.