Arsenic trioxide-induced apoptosis through oxidative stress in cells of colon cancer cell lines

Yoshihito Nakagawa, Yukihiro Akao, Hiroshi Morikawa, Ichiro Hirata, Kenichi Katsu, Tomoki Naoe, Nobuko Ohishi, Kunio Yagi

Research output: Contribution to journalArticle

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Abstract

Exposure of three colon cancer cell lines, SW480, DLD-1, and COLO201, to arsenic trioxide in the medium induced a marked concentration-dependent suppression of cell growth. The intracellular contents of reduced glutathione (GSH) in these cell lines tended to be inversely correlated with the sensitivity of the cells to arsenic trioxide. Among the cell lines, SW480 cells underwent apoptosis at the low arsenic trioxide concentration of 2 μM, which was prevented by pretreatment of the cells with N-acetylcysteine and was enhanced by buthionine sulfoximine. The production of reactive oxygen intermediates which were examined by 2′,7′-dichlorodihydrofluorescein diacetate (H2DCF-DA), increased with time after treatment with arsenic trioxide. The apoptosis was executed by the activation of caspase 3, which was shown by Western blot, enzymatic activity, and apoptosis inhibition assay. The mitochondrial membrane potential of adherent apoptotic SW480 cells and the cells from intermediate layer separated by density gradient centrifugation, both of which showed the active form of caspase 3 by Western blot analysis, was not lost. The overexpression of Bcl-2 protein in SW480 cells could not prevent the apoptosis induced by the treatment with arsenic trioxide. All these findings indicate that arsenic trioxide-induced apoptosis in SW480 cells is executed by the activation of caspase 3 without mediating by mitochondria under the overproduction of reactive oxygen species.

Original languageEnglish
Pages (from-to)2253-2269
Number of pages17
JournalLife Sciences
Volume70
Issue number19
DOIs
Publication statusPublished - 29-03-2002

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Oxidative stress
Colonic Neoplasms
Oxidative Stress
Cells
Apoptosis
Cell Line
Caspase 3
Chemical activation
Buthionine Sulfoximine
Mitochondria
Centrifugation
Western Blotting
Acetylcysteine
Cell growth
Glutathione
Density Gradient Centrifugation
arsenic trioxide
Mitochondrial Membrane Potential
Assays
Reactive Oxygen Species

All Science Journal Classification (ASJC) codes

  • Pharmacology

Cite this

Nakagawa, Yoshihito ; Akao, Yukihiro ; Morikawa, Hiroshi ; Hirata, Ichiro ; Katsu, Kenichi ; Naoe, Tomoki ; Ohishi, Nobuko ; Yagi, Kunio. / Arsenic trioxide-induced apoptosis through oxidative stress in cells of colon cancer cell lines. In: Life Sciences. 2002 ; Vol. 70, No. 19. pp. 2253-2269.
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Nakagawa, Y, Akao, Y, Morikawa, H, Hirata, I, Katsu, K, Naoe, T, Ohishi, N & Yagi, K 2002, 'Arsenic trioxide-induced apoptosis through oxidative stress in cells of colon cancer cell lines', Life Sciences, vol. 70, no. 19, pp. 2253-2269. https://doi.org/10.1016/S0024-3205(01)01545-4

Arsenic trioxide-induced apoptosis through oxidative stress in cells of colon cancer cell lines. / Nakagawa, Yoshihito; Akao, Yukihiro; Morikawa, Hiroshi; Hirata, Ichiro; Katsu, Kenichi; Naoe, Tomoki; Ohishi, Nobuko; Yagi, Kunio.

In: Life Sciences, Vol. 70, No. 19, 29.03.2002, p. 2253-2269.

Research output: Contribution to journalArticle

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AU - Nakagawa, Yoshihito

AU - Akao, Yukihiro

AU - Morikawa, Hiroshi

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AU - Naoe, Tomoki

AU - Ohishi, Nobuko

AU - Yagi, Kunio

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N2 - Exposure of three colon cancer cell lines, SW480, DLD-1, and COLO201, to arsenic trioxide in the medium induced a marked concentration-dependent suppression of cell growth. The intracellular contents of reduced glutathione (GSH) in these cell lines tended to be inversely correlated with the sensitivity of the cells to arsenic trioxide. Among the cell lines, SW480 cells underwent apoptosis at the low arsenic trioxide concentration of 2 μM, which was prevented by pretreatment of the cells with N-acetylcysteine and was enhanced by buthionine sulfoximine. The production of reactive oxygen intermediates which were examined by 2′,7′-dichlorodihydrofluorescein diacetate (H2DCF-DA), increased with time after treatment with arsenic trioxide. The apoptosis was executed by the activation of caspase 3, which was shown by Western blot, enzymatic activity, and apoptosis inhibition assay. The mitochondrial membrane potential of adherent apoptotic SW480 cells and the cells from intermediate layer separated by density gradient centrifugation, both of which showed the active form of caspase 3 by Western blot analysis, was not lost. The overexpression of Bcl-2 protein in SW480 cells could not prevent the apoptosis induced by the treatment with arsenic trioxide. All these findings indicate that arsenic trioxide-induced apoptosis in SW480 cells is executed by the activation of caspase 3 without mediating by mitochondria under the overproduction of reactive oxygen species.

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