title: Tobacco particulate matter is more potent than nicotine at upregulating nicotinic receptors on SH-SY5Y cells.
abstract: The effect of total particulate matter (TPM) from cigarette smoke on the expression and binding properties of nicotinic acetylcholine receptors (nAChRs) was investigated using a human neuroblastoma cell line (SH-SY5Y). TPM but not nicotine on its own inhibited cell growth at nicotine concentrations above 5 microM. To examine effects on nAChR expression, intact cells were incubated with 3H-epibatidine, and a Bmax of 13 fmoles/10(5) cells (7.8 x 10(4) binding sites/cell) was measured in unexposed cells as well as in cells treated with 2 microM nicotine alone or with TPM containing 2 microM nicotine. Using Scatchard analysis, we measured a Kd of 0.3 nM for 3H-epibatidine binding to nAChRs. This Kd was increased to 1.3 nM by addition of nicotine or TPM extract, both at 2 microM nicotine. Bmax, however, was unaffected, suggesting competitive binding of nicotine to its receptor. Short-term and prolonged 3-day exposures of SH-SY5Y cells to either TPM or nicotine at nicotine concentrations ranging from 0.2 microM to 20 microM increased specific binding, suggesting upregulation of nAChR expression. Most significant, binding was consistently greater in cells pretreated with TPM than in cells pretreated with nicotine. We conclude that TPM contains compounds that are toxic to cells at high concentrations (cell growth inhibition) but that do not compete with nicotine for binding to nAChRs (Scatchard analysis). These non-nicotinic compounds are capable of increasing the expression of one or more of the nAChR subunits. Furthermore, our cell culture assay provides a useful in vitro model for assessing the relative addictiveness of different tobacco products, including that of non-nicotine components.
authors.affiliation: Institute of Environmental Science and Research Ltd., Porirua, New Zealand.
authors.forename: Vikki
authors.initials: V
authors.lastname: Ambrose
authors.forename: John H
authors.initials: JH
authors.lastname: Miller
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Dickson
authors.forename: Scott
authors.initials: S
authors.lastname: Hampton
authors.forename: Penelope
authors.initials: P
authors.lastname: Truman
authors.forename: Rodney A
authors.initials: RA
authors.lastname: Lea
authors.forename: Jefferson
authors.initials: J
authors.lastname: Fowles
mesh_terms: D001665:Binding Sites; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects; D009447:Neuroblastoma; D009538:Nicotine / Q000494:pharmacology*; D018722:Nicotinic Agonists / Q000494:pharmacology*; D018408:Patch-Clamp Techniques; D011978:Receptors, Nicotinic / Q000187:drug effects* / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects*
chemical_list: D018722:Nicotinic Agonists; D011978:Receptors, Nicotinic; D009538:Nicotine