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Antiproliferative properties of lipids extracted from grain sorghum (GS) dry distiller’s grain (DDG) were analyzed to determine the feasibility of developing GS coproducts as a source for human health dietary ingredients. The lipid extract of GSDDG was delivered to human colon carcinoma (Caco-2) cells by solubilizing 0−1000 μg/mL of GS-DDG lipids in 100 μg/mL increments with micelles. A significant reduction in cell viability (25−50%) resulted at treatment levels of 400−1000 μg/mL GS-DDG lipids (p < 0.05). Alternatively, total protein levels of cells treated with 400, 500, and 600 μg/mL of GS-DDG lipid were not significantly different from the control, indicating cell growth during the treatment period. Total cell counts for the control were not significantly different from the GS-DDG lipid treated cells, but dead cell counts increased by 10% for the latter sample with a concomitant increase of the intercellular protein lactate dehydrogenase leakage (30−40%) in the medium. Preliminary analysis by the fluorescence-activated cell method (FACs) demonstrated that nonviable cells were in either the early apoptotic, late apoptotic, or necrotic stage post-treatment with 400, 500, and 600 μg/mL GS-DDG lipids. Physiochemical characterization of the GS-DDG lipids used for the antiproliferation study showed the presence of vitamin E (predominantly γ-tocopherol), triacylglycerides (predominantly linoleic acid), policosanols, aldehydes, and sterols (predominantly campesterol and stigmasterol), each of which or as synergistic/additive group of constituents may be responsible for the antiproliferative effect.