Previous in-vitro studies with progesterone (P), curcumin (cur), and vitamins gave the idea for a combination treatment of human melanoma cells. The aim was to study the combined effects of curcumin, vitamins (D3 and A), and steroids (P4 and RU-486) on human melanoma (BLM, 1205Lu, WM238) cell growth and IL-8 secretion in vitro. Human melanoma cells were incubated in a 96 well plate with a single (solo) compound or a double combo of the compounds or a triple combo of the compounds in various combinations. Supernatants were subjected to Elisarray and quantitation of the cytokine IL-8 secreted by the cells. Results from the three cell lines (BLM, 1205Lu and WM238) were compared. Solo or single compound treatment of Cur, P, RU, A and D3 resulted in 40 to 57% of BLM cell growth compared to its untreated control cell growth at 100%. Similarly, 1205Lu cell line treatments resulted in 37 to 59% growth compared to its untreated control cell growth at 100%. A double combo of compounds such as Cur+D3, Cur+P, treatments resulted in 19 to 42% of BLM cell growth and 17 to 37% of 1205Lu cell growth. Whereas WM238 cell line showed an 18 – 22% cell growth. A triple combo such as Cur+P+RU, treatments resulted in 21 to 30% of BLM cell growth and 19 to 30% of 1205Lu cell growth. Whereas WM238 cell line resulted in 19 to 20% cell growth. Elisarray of the supernatants pointed to the following two combo treatments A+RU, Cur+P+RU as having a basal secretion profile of various proinflammatory cytokines by the three cell lines with IL-8 as the only regulated cytokine. Hence, IL-8 in the supernatants of the two combo treatments was quantitated by Elisa. IL-8 secretion by the BLM cell line was 46.1 pg/ml and 31.08 pg/ml respectively in the supernatants of the two combos A+RU and Cur+P+RU. Whereas IL-8 secretion by the 1205Lu cell line was 17.5 and 8.0 pg/ml respectively and IL-8 secretion by the WM238 cell line was 0.515 and 0.926 pg/ml respectively. Conclusion: Measurements based on cell growth, Elisarray and IL-8 quantitation indicated the two combos A+RU and Cur+P+RU fared well on the three melanoma cell lines, suggesting a combination of compounds was more effective than the individual treatment of a compound.

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