Mechanism underlying Chios gum mastic-induced apoptosis on SCC25 human tongue squamous cell carcinoma cell line
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Chios gum mastic (CGM) is a resin produced from thestem and leaves of Pistiacia lentiscus L var chia, a plantwhich grows only on Chios Island in Greece. CGM has beenused for many centuries as a dietary supplement and folkmedicine for stomach and duodenal ulcers in manyMediterranean countries and is known also to induce cellcycle arrest and apoptosis in some cancer cells. In this study,we further investigated the induction and mechanismsunderlying the apoptotic response to CGM treatment in theSCC25 human tongue squamous cell carcinoma cell line. The viability of SCC25 cells, human normal keratinocytes(HaCaT cells) and human gingival fibroblasts (HGF-1cells), and the growth inhibition of SCC25 cells wereassessed by MTT assay and clonogenic assay, respectively. Staining with Hoechst and hemacolor dyes and TUNELassays were employed to detect SCC25 cells undergoingapoptosis. SCC25 cells were treated with CGM, and thiswas followed by western blotting, immunocytochemistry,confocal microscopy, FACScan flow cytometry, MMPactivity and proteasome activity analyses. CGM treatmentof SCC25 cells was found to result in a time- and dosedependentdecrease in cell viability, a dose-dependentinhibition of cell growth, and apoptotic cell death. Interestingly,CGM showed a remarkable level of cytotoxicity inSCC25 cells but not in normal cells. Tested SCC25 cells alsoshowed several lines of apoptotic manifestation. Takentogether, our present findings demonstrate that CGM stronglyinhibits cell proliferation by modulating the expression of G1cell cycle-related proteins and induces apoptosis via theproteasome, mitochondria and caspase cascades in SCC25 cells.
Abstract
Chios gum mastic (CGM) is a resin produced from thestem and leaves of Pistiacia lentiscus L var chia, a plantwhich grows only on Chios Island in Greece. CGM has beenused for many centuries as a dietary supplement and folkmedicine for stomach and duodenal ulcers in manyMediterranean countries and is known also to induce cellcycle arrest and apoptosis in some cancer cells. In this study,we further investigated the induction and mechanismsunderlying the apoptotic response to CGM treatment in theSCC25 human tongue squamous cell carcinoma cell line. The viability of SCC25 cells, human normal keratinocytes(HaCaT cells) and human gingival fibroblasts (HGF-1cells), and the growth inhibition of SCC25 cells wereassessed by MTT assay and clonogenic assay, respectively. Staining with Hoechst and hemacolor dyes and TUNELassays were employed to detect SCC25 cells undergoingapoptosis. SCC25 cells were treated with CGM, and thiswas followed by western blotting, immunocytochemistry,confocal microscopy, FACScan flow cytometry, MMPactivity and proteasome activity analyses. CGM treatmentof SCC25 cells was found to result in a time- and dosedependentdecrease in cell viability, a dose-dependentinhibition of cell growth, and apoptotic cell death. Interestingly,CGM showed a remarkable level of cytotoxicity inSCC25 cells but not in normal cells. Tested SCC25 cells alsoshowed several lines of apoptotic manifestation. Takentogether, our present findings demonstrate that CGM stronglyinhibits cell proliferation by modulating the expression of G1cell cycle-related proteins and induces apoptosis via theproteasome, mitochondria and caspase cascades in SCC25 cells.
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