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Gamma‐tocopherol induces apoptosis in cultured human salivary gland cancer cells

In the study reported here, we investigated both the possible antiproliferative and proapoptotic effects of either alpha‐tocopherol or gamma‐tocopherol in vitro upon a human salivary gland cancer cell line (ACC3). Cells were maintained in culture in RPMI media with added fetal bovine serum, calf serum, antibiotics, glutamine and fungicide. When ready for experimental use, cells were grown in a final concentration of 10, 25 or 50 micromolar of either alpha‐tocopherol or gamma‐tocopherol, and evaluated at 4 or 7 days. Cells were analyzed for their proliferative activity by cell counts and MTT assay. The presence of apoptosis was detected by DNA extraction, followed by size fractionation to look for DNA laddering. Apoptosis in cells was also detected and quantified using a TUNEL reaction mixture from Roche Diagnostics, followed by image analysis of cells using a fluorescence microscope. Cell counts and MTT assays showed that both test compounds had an inhibitory effect upon the growth of ACC3 cells; however, DNA size analysis, TUNEL assay and fluorescence image analysis indicated that apoptosis was induced by gamma‐tocopherol but not by alpha‐tocopherol.