Int J Oral Biol.  2011 Sep;36(3):155-162.

Apoptotic Effects of A Cisplatin and Eugenol Co-treatment of G361 Human Melanoma Cells

Affiliations
  • 1Department of Oral Anatomy, School of Dentistry, Pusan National University, Korea. parkbs@pusan.ac.kr

Abstract

Eugenol (4-allyl-2-methoxyphenol) is a naturally occurring phenolic compound that is widely used in dentistry as a component of zinc oxide eugenol cement that is commonly applied to the mouth environment. Cisplatin is one of the most potent known anticancer agents and shows significant clinical activity against a variety of solid tumors. This study was undertaken to investigate the synergistic apoptotic effects of co-treatments with eugenol and cisplatin on human melanoma (G361) cells. To investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment, an MTT assay was conducted. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and an analysis of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following this co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed. The results indicated that a co-treatment with eugenol and cisplatin induced multiple pathways and processes associated with an apoptotic response in G361 cells including nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, the increase and decrease of Bax and Bcl-2, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of 300 microM eugenol or 3 microM cisplatin for 24 h did not induce apoptosis. Our present data thus suggest that a combination therapy of eugenol and cisplatin is a potential treatment strategy for human melanoma.

Keyword

eugenol; cisplatin; apoptosis; G361 human melanoma cell line

MeSH Terms

Antineoplastic Agents
Apoptosis
Blotting, Western
Caspase 3
Caspase 7
Caspase 9
Cisplatin
Cytochromes c
Cytosol
Dentistry
DNA
DNA Fragmentation
Electrophoresis
Eugenol
Humans
Melanoma
Membrane Potential, Mitochondrial
Mouth
Phenol
Proteasome Endopeptidase Complex
Proteins
Zinc Oxide-Eugenol Cement
Antineoplastic Agents
Caspase 3
Caspase 7
Caspase 9
Cisplatin
Cytochromes c
DNA
Eugenol
Phenol
Proteasome Endopeptidase Complex
Proteins
Zinc Oxide-Eugenol Cement
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