Insights into Cadmium-Induced Carcinogenesis through an In Vitro Study Using C3H10T1/2Cl8 Cells: The Multifaceted Role of Mitochondria.
Monica OldaniAnna Maria VillaMarta ManzoniPasquale MelchiorettoPaolo ParentiEugenio MontiPaola FusiMatilde Emma ForcellaChiara UraniPublished in: International journal of molecular sciences (2021)
In this paper, we report the metabolic characterization of two foci, F1 and F3, obtained at the end of Cell Transformation Assay (CTA), performed by treating C3H10T1/2Cl8 mouse embryo fibroblasts with 1 μM CdCl2 for 24 h. The elucidation of the cadmium action mechanism can be useful both to improve the in vitro CTA and to yield insights into carcinogenesis. The metabolism of the two foci was investigated through Seahorse and enzyme activity assays; mitochondria were studied in confocal microscopy and reactive oxygen species were detected by flow cytometry. The results showed that F1 focus has higher glycolytic and TCA fluxes compared to F3 focus, and a more negative mitochondrial membrane potential, so that most ATP synthesis is performed through oxidative phosphorylation. Confocal microscopy showed mitochondria crowded in the perinuclear region. On the other hand, F3 focus showed lower metabolic rates, with ATP mainly produced by glycolysis and damaged mitochondria. Overall, our results showed that cadmium treatment induced lasting metabolic alterations in both foci. Triggered by the loss of the Pasteur effect in F1 focus and by mitochondrial impairment in F3 focus, these alterations lead to a loss of coordination among glycolysis, TCA and oxidative phosphorylation, which leads to malignant transformation.
Keyphrases
- reactive oxygen species
- cell death
- flow cytometry
- oxidative stress
- endoplasmic reticulum
- high glucose
- heavy metals
- high throughput
- diabetic rats
- cell cycle arrest
- protein kinase
- mesenchymal stem cells
- signaling pathway
- risk assessment
- bone marrow
- endothelial cells
- climate change
- mass spectrometry
- human health
- combination therapy