Benzyl Isothiocyanate Induces Growth Inhibition and Cell Apoptosis in Human Melanoma A375.S2 Cells

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Title:	Benzyl Isothiocyanate Induces Growth Inhibition and Cell Apoptosis in Human Melanoma A375.S2 Cells
Authors:	Liu-Wei Wu
Contributors:	Department of Biotechnology
Keywords:	Benzyl Isothiocyanate, Melanoma, Apoptosis
Date:	2010
Issue Date:	2010-11-08 01:10:44 (UTC+0)
Publisher:	Asia University
Abstract:	Skin cancer is one of the common carcinomatous diseases. It is becoming increasingly common in younger populations.Recently, many reports indicated that increased intake of cruciferous vegetables may be effective in preventing the risk of cancer and to reduce the incidence of cancer. Benzyl isothiocyanate (BITC), a compound presented in cruciferous vegetables, had shown to induce cell cycle arrest and apoptosis in many cancer cells. There is no report to show BITC inhibited the growth of A375.S2 human skin cancer cells. In this study BITC affecting apoptosis in A375.S2 cells was investigated. MTT assay was used to measure cell viability of A375.S2 cells after BITC treatment. DNA damage was determined by DNA fragmentation assay, DAPI staining and Comet assay. Flow cytometric analysis was performed to investigate the levels of mitochondrial membrane potential (ΔΨm), the production of reactive oxygen species (ROS), intracellular Ca2+ release and cell cycle distribution. Finally, we used the flow cytometry to examine caspase-3 activity and Annexin V affinity assay for apoptosis. In western blotting assay, we found cytochrome c, AIF and Endo G were released from mitochondria and activated downstream pathway to cause cell apoptosis. Our results showed that BITC treatment for 24 h significantly reduced the cell survival with an IC50 of 10±0.5 μM. BITC induced cell cycle arrest at G2/ M phase in A375.S2 cells. Moreover, BITC also caused DNA damage, decreased ΔΨm and increased ROS and intracellular Ca2+ levels. These observations indicate that BITC could induce apoptosis in human melanoma A375.S2 cell.
Appears in Collections:	[Department of Biotechnology] Theses & dissertations

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