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Orthotopic Implantation and Peripheral Immune Cell Monitoring in the II-45 Syngeneic Rat Mesothelioma Model
Published on: October 2, 2015
Volume-regulated Cl- channels in human pleural mesothelioma cells
Giuliano Meyer1, Simona Rodighiero, Fabiana Guizzardi
1Department of Biomolecular Sciences and Biotechnologies, University of Milan, Via Celoria 26, I-20133 Milan, Italy. giuliano.meyer@unimi.it
Abstract:
Anion channels in human mesothelial and mesothelioma cell lines were characterized by patch-clamp and biomolecular approaches. We found an outwardly rectifying anionic current which was inactivated at positive voltages and inhibited by extracellular adenosine 5'-triphosphate (ATP). Mesothelial and mesothelioma cells behaved differently concerning current inactivation properties. Inactivation is more pronounced and has a steeper onset in mesothelial cells. Different reversal potentials, in asymmetrical Cl(-) solutions, that could be attributed to a different selectivity of the channel, have been observed in the two cell lines. Mesothelioma cell single-channel analysis indicates that the number of the same active anion channel (3-4 pS) increased under hypoosmotic conditions. Immunocytochemistry experiments showed the presence of ICln protein in the cytosol and in the plasma membrane. Western blot analysis revealed an increase of ICln in the membrane under hypotonic conditions, an event possibly related to the activation of Cl(-) channels.
Insights
Human mesothelial and mesothelioma cells exhibit distinct anion channel properties, with differences in current inactivation and selectivity. These findings shed light on cellular chloride channel regulation.
Area of Science:
- Cell Biology
- Ion Channel Physiology
- Cancer Research
Background:
- Anion channels play critical roles in cellular function, including volume regulation and membrane potential maintenance.
- Mesothelial cells form a protective lining, while mesothelioma is a cancer arising from these cells.
- Understanding ion channel differences between normal and cancerous cells is crucial for therapeutic development.
Purpose of the Study:
- To characterize and compare anion channel properties in human mesothelial and mesothelioma cell lines.
- To investigate the role of adenosine 5'-triphosphate (ATP) in modulating these channels.
- To explore the involvement of ICln protein in anion channel activity.
Main Methods:
- Patch-clamp electrophysiology to record anionic currents.
- Biomolecular techniques including immunocytochemistry and Western blot analysis.
- Assessment of channel behavior under varying voltage, ionic, and osmotic conditions.
Main Results:
- An outwardly rectifying anionic current, inactivated at positive voltages and inhibited by extracellular ATP, was identified.
- Distinct inactivation properties and reversal potentials were observed between mesothelial and mesothelioma cells, suggesting differences in channel selectivity.
- Hypoosmotic conditions increased the number of active 3-4 pS anion channels in mesothelioma cells.
- ICln protein was detected in both cell types, with increased membrane presence under hypotonic conditions.
Conclusions:
- Human mesothelial and mesothelioma cells possess distinct anion channel characteristics, particularly in current inactivation and selectivity.
- Extracellular ATP modulates anion channel activity, with differential effects observed between cell types.
- The ICln protein is implicated in anion channel regulation, potentially mediating responses to osmotic stress.
