title: Cytotoxicity of chronic exposure to 4 cigarette smoke condensates in 2 cell lines.
abstract: Tobacco use is the leading preventable cause of death. The cytotoxicity of cigarette smoke condensate (CSC), the particulate fraction of cigarette smoke without the vapor phase, has mostly been tested in short-term in vitro studies lasting from a few hours to a few days. Here, we assessed the toxicity of CSCs from 2 reference cigarettes, 3R4F and CM6, using a primary human small airway epithelial (PSAE) cell line by quantifying adenosine 5'-triphosphate (ATP), 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxy-methoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS), total glutathione (reduced glutathione [GSH] + oxidized glutathione [GSSG]), and lactate dehydrogenase (LDH) release over the course of 28 days. The CSCs, 0.3 to 10 μg/mL, promoted cell proliferation at 120 hours of exposure, but demonstrated cytotoxicity at days 14 and 28. Interestingly, CSCs, 0.3 to 3 μg/mL, showed a cell death effect at day 14 but induced cell proliferation at day 28. Consistently, transformation associated with morphological changes began by day 14 and the transformed cells grew dramatically at day 28. The LDH assay appeared to be sensitive for assessing early cell damage, whereas the ATP, MTS, and GSH assays were more suitable for determining later stage CSCs-induced cytotoxicity. The ATP assay showed greater sensitivity than the MTS and GSH assays. We also assessed the toxicity of CSCs in an human Telomerase Reverse Transcriptase (hTERT)-immortalized Barrett esophagus cell line (CP-C). The CP-C cells demonstrated dose- and time-dependent cytotoxicity over the course of 28 days but displayed higher resistance to CSCs than PSAE cells. This study demonstrates that CSCs cause cytotoxicity and induce transformation related to cell resistance and cell invasion properties.
authors: Division of Biochemical Toxicology, FDA/National Center for Toxicological Research, Jefferson, AR, USA.
authors: Honggang
authors: H
authors: Wang
authors: Division of Biochemical Toxicology, FDA/National Center for Toxicological Research, Jefferson, AR, USA.
authors: Beverly
authors: B
authors: Word
authors: Division of Biochemical Toxicology, FDA/National Center for Toxicological Research, Jefferson, AR, USA.
authors: Lascelles
authors: L
authors: Lyn-Cook
authors: Office of Science, FDA/Center for Tobacco Products, Rockville, MD, USA.
authors: Maocheng
authors: M
authors: Yang
authors: Division of Biochemical Toxicology, FDA/National Center for Toxicological Research, Jefferson, AR, USA.
authors: George
authors: G
authors: Hammons
authors: Division of Biochemical Toxicology, FDA/National Center for Toxicological Research, Jefferson, AR, USA beverly.lyn-cook@fda.hhs.gov.
authors: Beverly
authors: B
authors: Lyn-Cook
mesh_terms: D000255:Adenosine Triphosphate / Q000032:analysis; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000032:analysis; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D020545:Respiratory Mucosa / Q000737:chemistry / Q000187:drug effects*; D012907:Smoking / Q000009:adverse effects*; D013778:Tetrazolium Salts / Q000032:analysis; D013844:Thiazoles / Q000032:analysis
chemical_list: D013778:Tetrazolium Salts; D013844:Thiazoles; C070380:3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium; D000255:Adenosine Triphosphate; D007770:L-Lactate Dehydrogenase; D005978:Glutathione
keywords: PSAE cells; cell-specific responses; chronic exposure; cigarette smoke condensate; hTERT cells; in vitro cytotoxicity assay