Effects of imipramine on ion channels and proliferation of IGR1 melanoma cells.
Human IGR1 cells are a model for malignant melanoma. Since progression through the cell cycle is accompanied by transient cell hyperpolarization, we studied the properties of potassium and chloride ion channels and their impact on cell growth. The major potassium current components were mediated by outward rectifying ether à go-go (hEAG) channels and Ca2+-activated channels (KCa) of the IK/SK type. The major chloride channel component was activated by osmotic cell swelling (Clvol). To infer about the contribution of these channels to proliferation, specific inhibitors are required. Since there is no specific blocker for hEAG available, we used the tricyclic antidepressant imipramine, which blocked all channels mentioned, in combination with blockers for KCa (charybdotoxin) and Clvol (DIDS and pamoic acid). Incubation of IGR1 cells for 48 hr in 10-15 mM imipramine reduced DNA synthesis and metabolism without significant effects on apoptosis. hEAG channels were most sensitive to imipramine (IC50: 3.4 microM at +50 mV), followed by KCa (13.8 microM at +50 mV) and Clvol (12 microM at -100 mV), indicating that hEAG expression may be of importance for proliferation of melanoma cells. The contribution of KCa channels could be excluded, as 500 nM charybdotoxin, which completely blocked KCa, had no effect on proliferation. The impact of Clvol also seems to be minor, because 500 microM pamoic acid, which completely blocked Clvol, did not affect proliferation either.
Gavrilova-Ruch
O
O
Research Unit Molecular and Cellular Biophysics, Medical Faculty of the Friedrich Schiller University Jena, Drackendorfer Strasse 1, D-07747 Jena, Germany.
Schönherr
K
K
Gessner
G
G
Schönherr
R
R
Klapperstück
T
T
Wohlrab
W
W
Heinemann
S H
SH
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002455:Cell Division / Q000187:drug effects; D018999:Charybdotoxin / Q000494:pharmacology; D018118:Chloride Channels / Q000187:drug effects / Q000502:physiology; D004247:DNA; D006801:Humans; D007099:Imipramine / Q000494:pharmacology*; D015640:Ion Channel Gating / Q000502:physiology; D007473:Ion Channels; D008546:Melanoma, Experimental / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009284:Naphthols / Q000494:pharmacology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000187:drug effects* / Q000502:physiology; D024681:Potassium Channels, Calcium-Activated / Q000037:antagonists & inhibitors* / Q000502:physiology; D014407:Tumor Cells, Cultured
D018118:Chloride Channels; D007473:Ion Channels; D009284:Naphthols; D015221:Potassium Channels; D024681:Potassium Channels, Calcium-Activated; D018999:Charybdotoxin; C004368:pamoic acid; D004247:DNA; D007099:Imipramine; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid