Int J Oral Biol.  2013 Sep;38(3):101-110.

Apoptotic Effects of Co-Treatment with a Chios Gum Mastic and Eugenol on G361 Human Melanoma Cells

• Jo JB
• Oh SH
• Kim IR
• Kim GC
• Kwak HH
• Park BS

Abstract

We investigated the synergistic apoptotic effects of co-treatments with Chios gum mastic (CGM) and eugenol on G361 human melanoma cells. An MTT assay was conducted to investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and analyses of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate expression and translocation of apoptosis-related proteins following CGM and eugenol co-treatment. Proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed.The results indicated that the co-treatment of CGM and eugenol induces multiple pathways and processes associated with an apoptotic response in G361 cells. These include nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, an increase of Bax and decrease of Bcl-2, a decreased DNA content, cytochrome c release 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 40 microg/ml CGM or 300 microM eugenol for 24 hours did not induce apoptosis. Our present data thus suggest that a combination therapy of CGM and eugenol is a potential treatment strategy for human melanoma.

Keyword

chios gum mastic; eugenol; apoptosis; human melanoma cells

MeSH Terms

Apoptosis
Blotting, Western
Caspase 3
Caspase 7
Caspase 9
Cytochromes c
Cytosol
DNA
DNA Fragmentation
Electrophoresis
Eugenol
Gingiva
Humans
Melanoma
Membrane Potential, Mitochondrial
Proteasome Endopeptidase Complex
Proteins
Resins, Plant
Caspase 3
Caspase 7
Caspase 9
Cytochromes c
DNA
Eugenol
Proteasome Endopeptidase Complex
Proteins
Resins, Plant