title: Methyl vinyl ketone, a toxic ingredient in cigarette smoke extract, modifies glutathione in mouse melanoma cells.
abstract: Cigarette smoke contains many harmful chemicals, which contribute to the pathogenesis of smoking-related diseases such as chronic obstructive pulmonary disease, cancer and cardiovascular disease. The cytotoxicity of cigarette smoke is well documented, but the definitive mechanism behind its toxicity remains unknown. Ingredients in cigarette smoke are known to deplete intracellular glutathione (GSH), the most abundant cellular thiol antioxidant, and to cause oxidative stress. In the present study, we investigated the mechanism of cigarette smoke extract (CSE)-induced cytotoxicity in B16-BL6 mouse melanoma (B16-BL6) cells using liquid chromatography-tandem mass spectrometry. CSE and ingredients in cigarette smoke, methyl vinyl ketone (MVK) and crotonaldehyde (CA), reduced cell viability in a concentration-dependent manner. Also, CSE and the ingredients (m/z 70, each) irreversibly reacted with GSH (m/z 308) to form GSH adducts (m/z 378) in cells and considerably decreased cellular GSH levels at concentrations that do not cause cell death. Mass spectral data showed that the major product formed in cells exposed to CSE was the GSH-MVK adduct via Michael-addition and was not the GSH-CA adduct. These results indicate that MVK included in CSE reacts with GSH in cells to form the GSH-MVK adduct, and thus a possible reason for CSE-induced cytotoxicity is a decrease in intracellular GSH levels.
authors.affiliation: School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women's University.
authors.forename: Shizuyo
authors.initials: S
authors.lastname: Horiyama
authors.forename: Yuta
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Mayuko
authors.initials: M
authors.lastname: Hatai
authors.forename: Chie
authors.initials: C
authors.lastname: Honda
authors.forename: Kiyoko
authors.initials: K
authors.lastname: Suwa
authors.forename: Atsushi
authors.initials: A
authors.lastname: Ichikawa
authors.forename: Noriko
authors.initials: N
authors.lastname: Yoshikawa
authors.forename: Kazuki
authors.initials: K
authors.lastname: Nakamura
authors.forename: Masaru
authors.initials: M
authors.lastname: Kunitomo
authors.forename: Sachiko
authors.initials: S
authors.lastname: Date
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Masujima
authors.forename: Mitsuo
authors.initials: M
authors.lastname: Takayama
mesh_terms: D000447:Aldehydes / Q000302:isolation & purification / Q000633:toxicity*; D000818:Animals; D002074:Butanones / Q000302:isolation & purification / Q000633:toxicity*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D003603:Cytotoxins / Q000302:isolation & purification / Q000633:toxicity*; D005978:Glutathione / Q000378:metabolism*; D051379:Mice; D012906:Smoke / Q000009:adverse effects / Q000032:analysis*; D012907:Smoking / Q000009:adverse effects / Q000378:metabolism
chemical_list: D000447:Aldehydes; D002074:Butanones; D003603:Cytotoxins; D012906:Smoke; C012796:2-butenal; C057920:3-buten-2-one; D005978:Glutathione