Our previous work on combo treatments of human melanoma (BLM) cell line pointed to the two combos A+RU and Cur+P+RU as the best combination to decrease cell growth, a basal secretion profile of pro-inflammatory cytokines and a decrease in the major pro-inflammatory cytokine IL-8 secretion by the cells. In order to confirm these findings, experiments were repeated on the 1205Lu cell line. Cells were incubated in a 96 well plate along with a combination of compounds for 48 hrs. After 48 hrs, cell growth was assayed using MTT and supernatants were collected for Elisarray and IL-8 quantitation by Elisa. Single or solo compound treatment resulted in cell growth in the range of 37 to 59% compared to untreated control 1205Lu cell growth at 100%. A double combo of compounds such as Cur+D3, Cur+P, Cur+RU, D3+P, D3+RU, P+RU, A+D3, A+P, A+RU resulted in cell growth in the range of 17 to 37%. A triple combo of compounds such as CUR+P+RU, Cur+A+D3, Cur+D3+RU resulted in cell growth in the range of 19 to 30%. Elisarray of the supernatants from various treatments pointed to A+RU, A+D3, Cur+D3+RU, Cur+P+RU as the combos having a basal secretion profile of pro-inflammatory cytokines. Quantitation of IL-8 by Elisa suggested A+RU and Cur+P+RU as the two combos having a minimal amount of IL-8 secretion 17.5 pg/ml and 8.0 pg/ml respectively.
Conclusion: These experiments suggested that the combination of compounds really decreased cell growth and IL-8 secretion. Two combos A+RU and Cur+P+RU were found to work well on both (BLM and 1205Lu) cell lines. These experiments will be repeated on other human melanoma cell lines for the confirmation of the above findings.
