RT Journal Article
T1 Knockout of PRDX6 induces mitochondrial dysfunction and cell cycle arrest at G2/M in HepG2 hepatocarcinoma cells.
A1 Lopez-Grueso, Maria Jose
A1 Lagal, Daniel Jose
A1 Garcia-Jimenez, Alvaro Fernando
A1 Tarradas, Rosa Maria
A1 Carmona-Hidalgo, Beatriz
A1 Peinado, Jose
A1 Requejo-Aguilar, Raquel
A1 Barcena, Jose Antonio
A1 Padilla, Carmen Alicia
K1 CRISPR-Cas9
K1 Carbohydrate metabolism
K1 Cell cycle
K1 Glucose metabolism
K1 Lipokines
K1 Mitochondria
K1 NRF2
K1 PCNA
K1 Peroxiredoxin 6
K1 Proteomics
K1 Redox proteome
AB Peroxiredoxin 6 (PRDX6) has been associated with tumor progression and cancer metastasis. Its acting on phospholipid hydroperoxides and its phospholipase-A2 activity are unique among the peroxiredoxin family and add complexity to its action mechanisms. As a first step towards the study of PRDX6 involvement in cancer, we have constructed a human hepatocarcinoma HepG2PRDX6-/- cell line using the CRISPR/Cas9 technique and have characterized the cellular response to lack of PRDX6. Applying quantitative global and redox proteomics, flow cytometry, in vivo extracellular flow analysis, Western blot and electron microscopy, we have detected diminished respiratory capacity, downregulation of mitochondrial proteins and altered mitochondrial morphology. Autophagic vesicles were abundant while the unfolded protein response (UPR), HIF1A and NRF2 transcription factors were not activated, despite increased levels of p62/SQSTM1 and reactive oxygen species (ROS). Insulin receptor (INSR), 3-phosphoinositide-dependent protein kinase 1 (PDPK1), uptake of glucose and hexokinase-2 (HK2) decreased markedly while nucleotide biosynthesis, lipogenesis and synthesis of long chain polyunsaturated fatty acids (LC-PUFA) increased. 254 Cys-peptides belonging to 202 proteins underwent significant redox changes. PRDX6 knockout had an antiproliferative effect due to cell cycle arrest at G2/M transition, without signs of apoptosis. Loss of PLA2 may affect the levels of specific lipids altering lipid signaling pathways, while loss of peroxidase activity could induce redox changes at critical sensitive cysteine residues in key proteins. Oxidation of specific cysteines in Proliferating Cell Nuclear Antigen (PCNA) could interfere with entry into mitosis. The GSH/Glutaredoxin system was downregulated likely contributing to these redox changes. Altogether the data demonstrate that loss of PRDX6 slows down cell division and alters metabolism and mitochondrial function, so that cell survival depends on glycolysis to lactate for ATP production and on AMPK-independent autophagy to obtain building blocks for biosynthesis. PRDX6 is an important link in the chain of elements connecting redox homeostasis and proliferation.
PB Elsevier
YR 2020
FD 2020-09-22
LK http://hdl.handle.net/10668/16387
UL http://hdl.handle.net/10668/16387
LA en
NO López-Grueso MJ, Lagal DJ, García-Jiménez ÁF, Tarradas RM, Carmona-Hidalgo B, Peinado J, et al. Knockout of PRDX6 induces mitochondrial dysfunction and cell cycle arrest at G2/M in HepG2 hepatocarcinoma cells. Redox Biol. 2020 Oct;37:101737
NO This research has been financed by grants from the Spanish Ministry of Economy and Competitiveness (BFU2016-80006-P) and the Andalusian Government (Consejería de Economía, Innovacion, ´ Ciencia y Empleo, BIO-0216). BC-H and RMT have been financed by Programa de Empleo Joven, FEDER/Junta de Andalucía, EJ17-BIO216 and EJI-17- BIO216, respectively. D.J.L. is recipient of a predoctoral fellowship Mod. 6.2–2018 from the University of Cordoba.
DS RISalud
RD Apr 12, 2025