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Reactive oxygen species as mediators of cell adhesion
1Universita' degli Studi di Firenze, Dipartimento di Scienze Biochimiche, Firenze, Italy. paola.chiarugi@unifi.it
Abstract:
The intracellular production of reactive oxygen species (ROS) has a fundamental importance in both cell proliferation and apoptosis induction. Moreover, many experimental and epidemiological evidence indicate that ROS contribute to the initiation and promotion of carcinogenesis, and that drugs or treatments aimed to reduce the tissue content of ROS can be chemopreventive and curative against cancer. Recently, important observations on the role of ROS as physiological regulators of intracellular signaling cascades activated by growth factors through their tyrosine-kinase receptors have shed new light on the possible mechanisms that can sustain the promoting activity of ROS. The downstream effect of ROS production is the reversible oxidation of proteins. Redox sensitive proteins include protein tyrosine phosphatases (PTPs) as the active-site cysteine is the target of specific oxidation, and this modification can be reversed by intracellular reducing agents. The reversible oxidation of PTPs family member was demonstrated firstly for PTP1B during EGF signaling and then for LMW-PTP and SHP-2 during PDGF stimulation. The inhibition exerted by ROS on tyrosine-phosphatases helps the propagation of RTK signals mediated by protein tyrosine phosphorylation, generally associated with the proliferative stimulus. Our new data are consistent with a model in which ROS take a role in integrin signaling, and in which synergistic activation of Rac-1 by growth factors and adhesion molecules translates in a critical increase of intracellular oxidants up to a threshold level where inhibition of the tyrosine phosphatase LMW-PTP takes place. In seeking for potential molecular mechanisms for oxidative signaling by integrins, we found that transient oxidation/inactivation of LMW-PTP, a known negative regulator of RTK signaling, occurred during fibroblast adhesion to matrix, with a kinetic which paralleled the generation of ROS. Moreover, overexpression of LMW-PTP in NIH-3T3 fibroblasts delayed cell attachment to the substrate. Finally, constitutively high levels of intracellular ROS, as are observed in cells expressing active Rac, would attenuate anchorage dependence for growth, by substituting for integrin signaling in non adherent cells.
Insights
Reactive oxygen species (ROS) regulate cell signaling by oxidizing protein tyrosine phosphatases (PTPs). This study reveals ROS
Area of Science:
- Cell Biology
- Biochemistry
- Cancer Research
Background:
- Reactive oxygen species (ROS) play critical roles in cell proliferation and apoptosis.
- ROS are implicated in cancer initiation and promotion, with ROS-reducing agents showing chemopreventive potential.
- ROS act as physiological regulators of signaling cascades initiated by growth factors via tyrosine-kinase receptors.
Purpose of the Study:
- To investigate the role of ROS in integrin signaling.
- To elucidate the molecular mechanisms underlying oxidative signaling mediated by integrins.
- To understand how ROS influence fibroblast adhesion and anchorage-dependent growth.
Main Methods:
- Investigated the synergistic activation of Rac-1 by growth factors and adhesion molecules.
- Monitored the transient oxidation and inactivation of LMW-PTP during fibroblast adhesion to the extracellular matrix.
- Assessed the impact of LMW-PTP overexpression on cell attachment kinetics in NIH-3T3 fibroblasts.
Main Results:
- Synergistic activation of Rac-1 leads to increased intracellular oxidants, causing LMW-PTP inhibition.
- Transient oxidation and inactivation of LMW-PTP occurred during fibroblast adhesion, paralleling ROS generation.
- Overexpression of LMW-PTP delayed fibroblast cell attachment to the substrate.
Conclusions:
- ROS are integral to integrin signaling, mediating fibroblast adhesion and potentially influencing anchorage dependence.
- Oxidative inactivation of LMW-PTP by ROS is a key mechanism in integrin-mediated signaling.
- Sustained high ROS levels may decouple anchorage dependence from integrin signaling, impacting cell growth.