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High Doses of Multiple Antioxidant Vitamins: Essential Ingredients in Improving the Efficacy of Standard Cancer Therapy

Center for Vitamins and Cancer Research (K.N.P., A.K., W.C.C.), Department of Radiology, School of Medicine, University of Colorado Health Sciences Center, Denver, CO
All India Institutes of Medical Sciences (V.K.), New Delhi, INDIA

View larger version (160K): [in a new window]   Fig. 1. Melanoma cells (105) were plated in tissue culture dishes (60 mm), and d--tocopheryl succinate (-TS) and sodium succinate plus ethanol were added to separate cultures 24 hours after plating. Drugs and medium were changed at 2 and 3 days after treatment. Photomicrographs were taken 4 days after treatment. Control cultures showed fibroblastic cells as well as round cells in clumps; a) cultures treated with ethanol (1%) and sodium succinate (5–6 µg/ml) also exhibited fibroblastic morphology with fewer round cells; b) -TS treated cultures c) 5 µg/mL, and d) 6 mg/mL, showed a dramatic change in morphology [20] x300.

View larger version (31K): [in a new window]   Fig. 2. Neuroblastoma cells (NBP2) were plated in tissue culture dishes (60 mm), and the cells were gamma-irradiated 24 hours after plating. Vitamin E succinate or the solvent (ethanol 0.25% and sodium succinate 5 µg/mL) was added immediately before irradiation. The drugs and medium were changed after 2 days of treatment. The number of cells per dish was determined after 3 days of treatment. Each experiment was repeated at least twice involving three samples per treatment. The average value (172±7x104) of untreated control NB cells was considered 100%, and the growth in treated cultures was expressed as a % of untreated controls. The bar at each point is standard error of the mean [41].

View larger version (21K): [in a new window]   Fig. 3. Neuroblastoma cells (50,000 per dish) were plated in tissue culture dishes (60 mm), and 5-Fluorouracil (5-FU) (0.08 µg/mL) plus sodium ascorbate or sodium ascorbate alone was added 24 hours after plating. The drug and medium were changed every day, and the number of cells per dish was determined 3 days after treatment. Each value represents the mean of six to nine samples±standard deviation [12].

View larger version (21K): [in a new window]   Fig. 4. Neuroblastoma cells (NBP2) (50,000 per dish) were plated in tissue culture dishes (60 mm), and vincristine and aqueous preparation of vitamin E (dl- tocopheryl acetate) were added 24 hours later. Drugs and medium were changed 2 days after treatment. The cell number and the number of trypan blue-stained cells were determined 3 days after treatment. The number of stained cells was subtracted from the total number of cells to obtain viable cells per dish. The average of control cultures was considered 100%. Each value represents an average of at least six samples. The bar of each point is standard deviation. The bars not shown in figure were equal to sizes of symbols [28].

View larger version (139K): [in a new window]   Fig. 5. Photomicrographs of neuroblastoma cells (NBP2) in culture after treatment with RO20-1724, an inhibitor of cyclic nucleotide phosphodiesterase, and a polar carotenoid originally referred to as beta-carotene [52]. Control a) 4 days after plating (50,000 cells/60-mm dish) showing mostly round cells; polar carotenoid (20 µg/mL-treated cells; b) 4 days after treatment showing no significant change in morphology; RO20-1724 (200 µg/mL)-treated cells; c) 4 days after treatment revealing increased number of cells with neurites; cells treated with RO20-1724 plus polar carotenoid d) for a period of 4 days showing more differentiated cells than those produced by RO20-1724 treatment alone; cells treated with RO20-1724 plus polar carotenoid for a period of 8 days e) and f) 11 days showing extensive network of neurites [53] x200.

View larger version (115K): [in a new window]   Fig. 6. Photomicrographs of murine melanoma (B16) cells were taken 4 days after treatment. Control culture contained cells with varied morphology (a). The melanoma cells treated with vitamin E succinate (4 µg/mL) were elongated, had long cytoplasmic processes, and were arranged alongside each other (b). Melanoma cells treated with RO20-1724, an inhibitor of cyclic nucleotide phosphodiesterase (100 µg/mL) were large and elongated, had some long processes, and were arranged alongside each other (c). A combination of RO20-1724 and vitamin E succinate (d) increased the level morphologic differentiation more than that produced by individual agents [54] x450.