title: The Effect of Cigarette Smoke on the Translocator Protein (TSPO) in Cultured Lung Cancer Cells.
abstract: Lung cancer is prevalent in cigarette smokers. The mitochondrial membrane translocator protein (TSPO), is thought to protect cells from free radical damage. We examined the effect of cigarette smoke (CS) (containing free radicals) alone and in the presence of saliva (containing redox active free iron), on survival of H1299 lung cancer cells and on their mitochondrial characteristics, and whether TSPO binding was influenced by CS and by saliva. We exposed H1299 cells to CS in the presence/absence of saliva and also characterized TSPO binding in the cells using [3H]PK 11195 as a radioligand. CS induced a significant drop in mitochondrial potential (ΔΨm), while addition of saliva did not lead to further loss of ΔΨm (42.5% vs. 39.85%). Scatchard analysis of the saturation curve of [3H]PK 11195 binding (0.2-6 nM final concentration) yielded a straight-line plot (R =  0.9). Average Bmax value was 3274 ± 787 fmol/mg of protein, and average Kd value was 9.2 ± 1.3 nM. Benzodiazepine diazepam partially prevented decrease in cell survival following exposure to CS and redox active iron containing media (saliva) while benzodiazepine clonazepam did not, indicating that this effect is TSPO-specific. Exposure of cells to CS resulted in alternation of biomolecules expressed by CLs peroxidation, reduction of TSPO binding, and depletion of the mitochondrial potential. This irreversible damage was enhanced in the presence of saliva. All these modulations may result in cellular death increase following CS exposure, enhanced in the presence of saliva.
authors.affiliation: Department of Neuroscience, Rappaport Family Institute for Research in the Medical Sciences, Technion-Israel Institute of Technology, 31096, Haifa, Israel.
authors.forename: Rafael
authors.initials: R
authors.lastname: Nagler
authors.affiliation: Department of Neuroscience, Rappaport Family Institute for Research in the Medical Sciences, Technion-Israel Institute of Technology, 31096, Haifa, Israel.
authors.forename: Shiri
authors.initials: S
authors.lastname: Cohen
authors.affiliation: Department of Neuroscience, Rappaport Family Institute for Research in the Medical Sciences, Technion-Israel Institute of Technology, 31096, Haifa, Israel.
authors.forename: Moshe
authors.initials: M
authors.lastname: Gavish
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D006801:Humans; D008175:Lung Neoplasms / Q000139:chemically induced / Q000235:genetics* / Q000473:pathology; D008928:Mitochondria / Q000187:drug effects / Q000473:pathology; D010084:Oxidation-Reduction / Q000187:drug effects; D018079:Receptors, GABA / Q000096:biosynthesis* / Q000235:genetics; D012463:Saliva / Q000187:drug effects / Q000378:metabolism; D012907:Smoking / Q000009:adverse effects*; D014026:Nicotiana / Q000009:adverse effects*
chemical_list: D018079:Receptors, GABA; C499229:TSPO protein, human
keywords: CELL SURVIVAL; CIGARETTE SMOKE; LUNG CANCER CELLS; TSPO