Proliferation of human lens epithelial cells (HLE-B3) is inhibited by blocking of voltage-gated calcium channels.
Calcium, as an integral part of a large number of cellular regulatory pathways, is selective in the control of specific cell functions like the start of G1 phase in cell cycle. Cell proliferation has been suggested to depend on increasing intracellular calcium levels. A major regulatory pathway for intracellular calcium is the calcium influx into the cell via voltage-gated calcium channels. T-type and L-type calcium channels are substantially present in human lens epithelial cell (hLEC), and total calcium currents are inhibited by mibefradil. Here, the hypothesis was tested if calcium influx via Ca(v) channels regulates proliferation in epithelial cells. Cell proliferation was determined by cell culture assays using the L- and T-type Ca(v) channel blockers mibefradil and verapamil as modulators for calcium influx. Calcium influx was investigated using the Manganese quench technique. Western blot experiments were accomplished under standard conditions using antibodies against MAPK 3. Mibefradil as well as verapamil impaired cell proliferation, but in different concentration ranges. Furthermore, the activation of MAPK 3 was reduced by both antagonists. Calcium influx was also reduced in the presence of both blockers. We conclude that the transmembrane influx of Ca(2+) through Ca(v) channels contributes to the regulation of hLEC proliferation, identifying Ca(v) channel blockers as potential therapeutic substances in ocular diseases.
Meissner
Anja
A
Institute of physiology, University of Rostock, Gertrudenstrasse 9, Rostock, 18055, Germany. anja.meissner@web.de
Noack
Thomas
T
D000465:Algorithms; D015153:Blotting, Western; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D015220:Calcium Channels / Q000187:drug effects* / Q000502:physiology*; D020746:Calcium Channels, L-Type / Q000187:drug effects / Q000502:physiology; D020747:Calcium Channels, T-Type / Q000187:drug effects / Q000502:physiology; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000502:physiology; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D004847:Epithelial Cells / Q000502:physiology*; D006801:Humans; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology; D007903:Lens Capsule, Crystalline / Q000166:cytology / Q000502:physiology; D007908:Lens, Crystalline / Q000166:cytology*; D020748:Mibefradil / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014700:Verapamil / Q000494:pharmacology
D002121:Calcium Channel Blockers; D015220:Calcium Channels; D020746:Calcium Channels, L-Type; D020747:Calcium Channels, T-Type; D020748:Mibefradil; D014700:Verapamil; D002118:Calcium