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Author Spotlight: Oxygen-Independent Assays to Measure Mitochondrial Function in Mammals
Published on: May 19, 2023
Mitochondrial DNA depletion causes decreased ROS production and resistance to apoptosis
Hulin Chen1, Junling Wang2, Zhongrong Liu1
1Department of Dermatology, Guangzhou General Hospital of Guangzhou Military Command (Liuhuaqiao Hospital), Guangzhou, Guangdong 510010, P.R. China.
Abstract:
Mitochondrial DNA (mtDNA) depletion occurs frequently in many diseases including cancer. The present study was designed in order to examine the hypothesis that mtDNA‑depleted cells are resistant to apoptosis and to explore the possible mechanisms responsible for this effect. Parental human osteosarcoma 143B cells and mtDNA‑deficient (Rho˚ or ρ˚) 206 cells (derived from 143B cells) were exposed to different doses of solar-simulated ultraviolet (UV) radiation. The effects of solar irradiation on cell morphology were observed under both light and fluorescence microscopes. Furthermore, apoptosis, mitochondrial membrane potential (MMP) disruption and reactive oxygen species (ROS) production were detected and measured by flow cytometry. In both cell lines, apoptosis and ROS production were clearly increased, whereas MMP was slightly decreased. However, apoptosis and ROS production were reduced in the Rho˚206 cells compared with the 143B cells. We also performed western blot analysis and demonstrated the increased release of cytosolic Cyt c from mitochondria in the 143B cells compared with that in the Rho˚206 cells. Thus, we concluded that Rho˚206 cells exhibit more resistance to solar‑simulated UV radiation‑induced apoptosis at certain doses than 143B cells and this is possibly due to decreased ROS production.
Insights
Mitochondrial DNA-depleted cells show increased resistance to ultraviolet radiation-induced apoptosis. This resistance in Rho˚206 cells is linked to reduced reactive oxygen species production compared to parental 143B cells.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- Mitochondrial DNA (mtDNA) depletion is common in various diseases, including cancer.
- Understanding the role of mtDNA in cellular responses to stress is crucial for disease research.
Purpose of the Study:
- To investigate if mtDNA-depleted cells exhibit resistance to apoptosis.
- To explore the underlying mechanisms of this potential resistance to apoptosis.
Main Methods:
- Comparison of parental 143B cells and mtDNA-deficient Rho˚206 cells.
- Exposure to solar-simulated ultraviolet (UV) radiation.
- Assessment of apoptosis, mitochondrial membrane potential (MMP), and reactive oxygen species (ROS) via flow cytometry and Western blot.
Main Results:
- UV radiation increased apoptosis and ROS production in both cell lines.
- Rho˚206 cells showed reduced apoptosis and ROS production compared to 143B cells.
- Lower release of cytosolic cytochrome c was observed in Rho˚206 cells.
Conclusions:
- mtDNA-deficient Rho˚206 cells are more resistant to UV-induced apoptosis than 143B cells.
- Decreased ROS production may explain the enhanced apoptosis resistance in mtDNA-depleted cells.
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