alpha-Bungarotoxin blocks the nicotinic receptor mediated increase in cell number in a neuroendocrine cell line.
Exposure of H69 small cell lung carcinoma cells to nicotinic agonists resulted in a significant increase (up to 100%) in cell number after 6 to 12 days. The effect of nicotine (10(-8) M to 10(-4) M) was both dose and time dependent as was that of another nicotinic agonist cytisine (10(-6) M to 10(-4) M). Interestingly, both the nicotine and cytisine induced increases in H69 cell number were blocked by alpha-bungarotoxin, as well as d-tubocurarine a nicotinic blocker which appears to interact with most nicotinic receptors. These results suggest that the nicotine induced increase in cell number is mediated through an interaction at the nicotinic alpha-bungarotoxin receptor. This idea is further supported by experiments which show (1) that H69 cells possess high affinity alpha-bungarotoxin sites (Kd = 25 nM, Bmax = 10.4 fmol/10(6) cells) with the characteristics of a nicotinic alpha-bungarotoxin receptor and (2) that the potencies of nicotinic receptor ligands in the alpha-bungarotoxin binding assay were similar to those observed in the functional studies. Northern analysis showed that mRNA for alpha 7, a putative nicotinic alpha-bungarotoxin binding subunit, and for alpha 5 were present in H69 cells. The present data provide further evidence that nicotine increases cell number in small cell lung carcinoma and are the first to show that this effect is mediated through an interaction at the nicotinic alpha-bungarotoxin receptor population. These results suggest that the alpha-bungarotoxin site may be involved in modulating proliferative responses in neuroendocrine derived SCLC cells.
Quik
M
M
Department of Pharmacology, McGill University, Montreal, Quebec, Canada.
Chan
J
J
Patrick
J
J
D000470:Alkaloids / Q000037:antagonists & inhibitors / Q000494:pharmacology; D001392:Azocines; D015152:Blotting, Northern; D002038:Bungarotoxins / Q000494:pharmacology*; D018288:Carcinoma, Small Cell / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D006801:Humans; D007457:Iodine Radioisotopes; D008175:Lung Neoplasms / Q000378:metabolism; D009490:Neurosecretory Systems / Q000166:cytology / Q000378:metabolism*; D009538:Nicotine / Q000037:antagonists & inhibitors / Q000494:pharmacology; D018733:Nicotinic Antagonists / Q000494:pharmacology*; D011807:Quinolizines; D012333:RNA, Messenger / Q000096:biosynthesis; D011978:Receptors, Nicotinic / Q000187:drug effects*; D014403:Tubocurarine / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D000093843:Quinolizidine Alkaloids
D000470:Alkaloids; D001392:Azocines; D002038:Bungarotoxins; D007457:Iodine Radioisotopes; D009538:Nicotine; D018733:Nicotinic Antagonists; D011807:Quinolizines; D012333:RNA, Messenger; D011978:Receptors, Nicotinic; D014403:Tubocurarine; C004712:cytisine; D000093843:Quinolizidine Alkaloids