Mitochondrial connexin 43 modulates metabolic stress adaptation in glioma cell lines

Gervasi, Anna (author), Potokar, Maja (author), Jorgačevski, Jernej (author), Zorec, Robert (author), Parenti, Rosalba (author), Vicario, Nunzio (author)

Background Connexin 43 (CX43) is a hemichannel (HC)- and gap junction (GJ)-forming protein that mediates the exchange of small molecules between the intracellular and extracellular environments, as well as intercellular communication. In addition to this canonical role, recent studies have shown that its functions range from transcriptional regulation to intracellular homeostasis. The ability of CX43 to translocate into mitochondria suggests its involvement in energy metabolism. However, the functions of mitochondrial CX43 (mt-CX43) in neural cells remain unexplored. Methods Our study investigated the expression and localisation of mt-CX43 through western blot and immunofluorescence analyses in four immortalised human glioma cell lines: T98-G, A-172, CCF-STTG1, and U-87 MG. Additionally, targeted metabolomic analysis was conducted to assess changes in key metabolic pathways. Results Basal CX43 expression and extracellular stress factors, particularly cell density and extracellular pH fluctuations, significantly modulated the mitochondrial localisation of CX43. Inhibition of the heat shock protein 90 (HSP90) chaperone system by geldanamycin (GA) resulted in a marked reduction in mt-CX43, suggesting an import mechanism involving HSP90 and the translocase of the outer membrane (TOM) complex. In addition, the assessment of key metabolites revealed increased purine biosynthesis in T98-G cells exposed to GA treatment, characterised by lower basal CX43 expression and reduced mt-CX43 levels under stress conditions. Conversely, U-87 MG cells exhibited a stable NAD$^+$/NADH ratio and a significant increase in NADH levels, indicating a metabolic shift towards a more resilient state. Conclusions Our results suggest that mt-CX43 serves as a multifunctional regulator of metabolic adaptation and stress response in glioma cell lines. Our results extend the role of mt-CX43 as an essential factor in cellular metabolic plasticity, providing new insights into the modulation of metabolic imbalances and mitochondrial dysfunction.

• Language: English
• Keywords: CX43, pH, geldanamycin, glioblastoma, metabolism, purine
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: MF - Faculty of Medicine
• Publication status: Published
• Publication version: Version of Record
• Year: 2025
• Number of pages: 19 str.
• Numbering: Vol. 23, iss. 1, art. 512
• PID: 20.500.12556/RUL-181447
• UDC: 577:61
• ISSN on article: 1478-811X
• DOI: 10.1186/s12964-025-02523-2
• COBISS.SI-ID: 260784899
• Publication date in RUL: 08.04.2026

Record is a part of a journal

• Title: Cell communication and signaling
• Shortened title: Cell commun. signal.
• Publisher: Springer Nature
• ISSN: 1478-811X
• COBISS.SI-ID: 513859097

Licences

• License: CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
• Link: http://creativecommons.org/licenses/by-nc-nd/4.0/
• Description: The most restrictive Creative Commons license. This only allows people to download and share the work for no commercial gain and for no other purposes.

Secondary language

• Language: Slovenian
• Keywords: geldanamicin, glioblastom, metabolizem, purin