Effect of mtDNA depletion from C6 glioma cells and characteristics of the generated C6ρ0 cells

Mol Med Rep. 2021 Apr;23(4):265. doi: 10.3892/mmr.2021.11904. Epub 2021 Feb 12.

Abstract

Malignant tumors of the central nervous system (CNS) are among the types of cancer with the poorest prognosis and glioma is the commonest primary CNS tumor. A mitochondrial DNA (mtDNA)‑depleted cell line C6ρ0 was generated from C6 glioma cells after long‑term exposure to ethidium bromide and 2',3'‑dideoxycytidine in order to determine the effect of mtDNA damage on cell proliferation and pathological changes in glioma cells. Single cell clones were isolated and identified after 42 days of incubation. Repopulated cybrids were formed when the clonal C6ρ0 cells were fused with rat platelets and no difference was observed in their growth in a selective medium without uridine and pyruvate compared with the growth of the parent C6 cells. Disruption of mtDNA resulted in changes in mitochondrial morphology, decreased cell proliferation, reduced intracellular reactive oxygen species and intracellular ATP, along with decreased mtDNA and mitochondrial membrane potential in C6ρ0 cells compared with the C6 cells. Taken together, C6ρ0 cells without mtDNA were established for the first time and their characteristics were compared with parent cells. This C6ρ0 cell line could be used to explore the contribution of mitochondrial dysfunction and mtDNA mutations in the pathogenesis of glioma.

Keywords: C6 glioma; ρ0; mitochondrial DNA; cytoplasmic hybrids.

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Brain Neoplasms / genetics*
  • Brain Neoplasms / metabolism
  • Brain Neoplasms / pathology
  • Cell Line, Tumor
  • Cell Proliferation / genetics
  • DNA, Mitochondrial / genetics*
  • DNA, Mitochondrial / metabolism
  • Glioma / genetics*
  • Glioma / metabolism
  • Glioma / pathology
  • Intracellular Space / metabolism
  • Membrane Potential, Mitochondrial / genetics
  • Membrane Potential, Mitochondrial / physiology
  • Mitochondria / genetics*
  • Mitochondria / metabolism
  • Rats
  • Reactive Oxygen Species / metabolism

Substances

  • DNA, Mitochondrial
  • Reactive Oxygen Species
  • Adenosine Triphosphate

Grants and funding

The present study was supported by grants from the National Science Foundation of China (grant no. 31870333), Qinghai Provincial Science Foundation (grant no. 2019-ZJ-7023), Qinghai Province International Cooperation Project (grant no. 2018-HZ-812), the Taishan Scholar Program of Shandong Province (grant no. tshw201502046) and the Shuangbai Project of Yantai and Youth Innovation Promotion Association, CAS.