Mechanism Underlying Shikonin-induced Apoptosis and Cell Cycle Arrest on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line
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Shikonin, a major ingredient in the traditional Chinese herbLithospermumerythrorhizon, exhibits multiple biologicalfunctions including antimicrobial, anti-inflammatory, andantitumor effects. It has recently been reported that shikonindisplays antitumor properties in many cancers. This study wasaimed to investigate whether shikonin could inhibit oralsquamous carcinoma cell (OSCC) growth via mechanisms ofapoptosis and cell cycle arrest. The effects of shikonin on theviability and growth of OSCC cell line, SCC25 cells wereassessed by MTT assay and clonogenic assays, respectively. Hoechst staining and DNA electrophoresis indicated that theshikonin-treated SCC25 cells were undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy,flow cytometry, MMP activity, and proteasome activity alsosupported the finding that shikonin induces apoptosis. Shikonin treatment of SCC25 cells resulted in a time- anddose-dependent decrease in cell viability, inhibition of cellgrowth, and increase in apoptotic cell death. The treatedSCC25 cells showed several lines of apoptotic manifestationas follows: nuclear condensation; DNA fragmentation;reduced MMP and proteasome activity; decrease in DNAcontents; release of cytochrome c into cytosol; translocation ofAIF and DFF40 (CAD) onto the nuclei; a significant shift inBax/Bcl-2 ratio; and activation of caspase-9, -7, -6, and -3, aswell as PARP, lamin A/C, and DFF45 (ICAD). Shikonintreatment also resulted in down-regulation of the G1 cellcycle-related proteins and up-regulation of p27KIP1. Takentogether, our present findings demonstrate that shikoninstrongly inhibits cell proliferation by modulating theexpression of the G1 cell cycle-related proteins, and that itinduces apoptosis via the proteasome, mitochondria, andcaspase cascades in SCC25 cells.
Abstract
Shikonin, a major ingredient in the traditional Chinese herbLithospermumerythrorhizon, exhibits multiple biologicalfunctions including antimicrobial, anti-inflammatory, andantitumor effects. It has recently been reported that shikonindisplays antitumor properties in many cancers. This study wasaimed to investigate whether shikonin could inhibit oralsquamous carcinoma cell (OSCC) growth via mechanisms ofapoptosis and cell cycle arrest. The effects of shikonin on theviability and growth of OSCC cell line, SCC25 cells wereassessed by MTT assay and clonogenic assays, respectively. Hoechst staining and DNA electrophoresis indicated that theshikonin-treated SCC25 cells were undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy,flow cytometry, MMP activity, and proteasome activity alsosupported the finding that shikonin induces apoptosis. Shikonin treatment of SCC25 cells resulted in a time- anddose-dependent decrease in cell viability, inhibition of cellgrowth, and increase in apoptotic cell death. The treatedSCC25 cells showed several lines of apoptotic manifestationas follows: nuclear condensation; DNA fragmentation;reduced MMP and proteasome activity; decrease in DNAcontents; release of cytochrome c into cytosol; translocation ofAIF and DFF40 (CAD) onto the nuclei; a significant shift inBax/Bcl-2 ratio; and activation of caspase-9, -7, -6, and -3, aswell as PARP, lamin A/C, and DFF45 (ICAD). Shikonintreatment also resulted in down-regulation of the G1 cellcycle-related proteins and up-regulation of p27KIP1. Takentogether, our present findings demonstrate that shikoninstrongly inhibits cell proliferation by modulating theexpression of the G1 cell cycle-related proteins, and that itinduces apoptosis via the proteasome, mitochondria, andcaspase cascades in SCC25 cells.
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