CFTR modulates programmed cell death by decreasing intracellular pH in Chinese hamster lung fibroblasts
| Autor: | Jean Michel Blasi, Laurent Counillon, Chantal Poujeol, Philippe Poujeol, Michel Tauc, Herve Barriere |
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| Přispěvatelé: | Physiologie cellulaire et moléculaire des systèmes intégrés (PCMSI), Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2001 |
| Předmět: |
MESH: Hydrogen-Ion Concentration
Physiology MESH: Cricetinae 8-Bromo Cyclic Adenosine Monophosphate Cystic Fibrosis Transmembrane Conductance Regulator Angiogenesis Inhibitors Apoptosis MESH: Iodides Cystic fibrosis MESH: Recombinant Proteins 0302 clinical medicine MESH: Cricetulus MESH: Reverse Transcriptase Polymerase Chain Reaction Cricetinae MESH: 8-Bromo Cyclic Adenosine Monophosphate MESH: Animals Lung MESH: Angiogenesis Inhibitors MESH: Cystic Fibrosis Transmembrane Conductance Regulator 0303 health sciences MESH: Kinetics MESH: Lovastatin biology Reverse Transcriptase Polymerase Chain Reaction MESH: Gluconates Hydrogen-Ion Concentration Recombinant Proteins Cystic fibrosis transmembrane conductance regulator Cell biology medicine.anatomical_structure Biochemistry 030220 oncology & carcinogenesis Programmed cell death Sodium-Hydrogen Exchangers Intracellular pH Hamster DNA Fragmentation Transfection Gluconates Chinese hamster Cell Line 03 medical and health sciences Cricetulus Chlorides Chloride Channels [SDV.MHEP.PHY]Life Sciences [q-bio]/Human health and pathology/Tissues and Organs [q-bio.TO] medicine MESH: DNA Fragmentation Animals MESH: Lung Lovastatin MESH: Chlorides Fibroblast 030304 developmental biology MESH: Sodium-Hydrogen Antiporter MESH: Transfection MESH: Apoptosis MESH: Chloride Channels Cell Biology Fibroblasts Iodides medicine.disease biology.organism_classification MESH: Cell Line Kinetics MESH: Fibroblasts Nitrobenzoates MESH: Nitrobenzoates biology.protein |
| Zdroj: | American Journal of Physiology-Cell Physiology American Journal of Physiology-Cell Physiology, American Physiological Society, 2001, 281 (3), pp.C810-24 ResearcherID |
| ISSN: | 1522-1563 0363-6143 |
| DOI: | 10.1152/ajpcell.2001.281.3.c810 |
| Popis: | To study the potential influence of cystic fibrosis conductance regulator (CFTR) on intracellular pH regulation during apoptosis induction, we used PS120 Chinese hamster lung fibroblasts devoid of the Na+/H+exchanger (NHE1 isoform) transfected with constructs, allowing the expression of CFTR and/or NHE1. Kinetics of lovastatin-induced apoptosis were measured by orcein staining, double staining with Hoechst-33258, propidium iodide, DNA fragmentation, and annexin V labeling. In PS120 control cells, the percentage of apoptotic cells after 40 h of lovastatin treatment was 23 ± 3%, whereas in PS120 CFTR-transfected cells, this percentage was 40 ± 4%. In PS120 NHE1 cells, the transfection with CFTR did not modify the percentage of apoptotic cells after 40 h (control: 19 ± 3%, n = 8; CFTR: 17 ± 1%, n = 8), indicating that blocking intracellular acidification by overexpressing the Na+/H+exchanger inhibited the enhancement of apoptosis induced by CFTR. In all cell lines, the initial pH values were identical (pH = 7.46 ± 0.04, n = 9), and treatment with lovastatin led to intracellular acidification. However, the pH value after 40 h was lower in PS120 CFTR-transfected cells (pH = 6.85 ± 0.02, n= 10) than in PS120 cells (pH = 7.15 ± 0.03, n = 10). To further investigate the origin of this increased intracellular acidification observed in CFTR-transfected cells, the activity of the DIDS-inhibitable Cl−/HCO[Formula: see text] exchanger was studied. 8-Bromoadenosine 3′,5′-cyclic monophosphate incubation resulted in Cl−/HCO[Formula: see text] exchanger activation in PS120 CFTR-transfected cells but had no effect on PS120 cells. Together, our results suggest that CFTR can enhance apoptosis in Chinese hamster lung fibroblasts, probably due to the modulation of the Cl−/HCO[Formula: see text] exchanger, resulting in a more efficient intracellular acidification. |
| Databáze: | OpenAIRE |
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