Methyl Vinyl Ketone, a Toxic Ingredient in Cigarette Smoke Extract, Modifies Glutathione in Mouse Melanoma Cells

Shizuyo Horiyama; Yuta Takahashi; Mayuko Hatai; Chie Honda; Kiyoko Suwa; Atsushi Ichikawa; Noriko Yoshikawa; Kazuki Nakamura; Masaru Kunitomo; Sachiko Date; Tsutomu Masujima; Mitsuo Takayama

doi:10.1248/cpb.c14-00109

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Methyl Vinyl Ketone, a Toxic Ingredient in Cigarette Smoke Extract, Modifies Glutathione in Mouse Melanoma Cells

Shizuyo Horiyama , Yuta Takahashi, Mayuko Hatai, Chie Honda, Kiyoko Suwa, Atsushi Ichikawa, Noriko Yoshikawa, Kazuki Nakamura, Masaru Kunitomo, Sachiko Date, Tsutomu Masujima, Mitsuo Takayama

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Shizuyo Horiyama Corresponding author
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Yuta Takahashi
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Department of Pharmacy, Gunma University Hospital
Mayuko Hatai
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Chie Honda
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Kiyoko Suwa
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Atsushi Ichikawa
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Noriko Yoshikawa
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Kazuki Nakamura
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Masaru Kunitomo
School of Pharmacy and Pharmaceutical Sciences, Mukogawa Women’s University
Sachiko Date
Riken Quantitative Biology Center, OLABB, Osaka University
Tsutomu Masujima
Riken Quantitative Biology Center, OLABB, Osaka University
Mitsuo Takayama
International Graduate School of Arts and Sciences, Yokohama City University

Keywords: cigarette smoke extract (CSE), glutathione (GSH), methyl vinyl ketone (MVK), B16-BL6 mouse melanoma (B16-BL6) cell, LC/MS, LC/MS/MS

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2014 Volume 62 Issue 8 Pages 772-778

DOI https://doi.org/10.1248/cpb.c14-00109

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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.

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