The In-Vitro Effect of Curcumin and Arsenic Trioxide on The Level Of NF-kB and Induction of Apoptosis in B16 Cell Lines

Abstract

Background: Malignant Melanoma (MM) is one of aggressive skin tumors that had increase in incidence during the past two decades. Curcumin is a natural product that had been utilized for a variety of maladies like rheumatic, cardiac, neurological and numerous other disorders while arsenic trioxide ‘ArT’ (not approved for treatment of MM) is well known cytotoxic agent. NF-kB is a specific complicated pertinacious element that participates in controlling of DNA transcription, regulating cytokine expression and, hence, cell survival. It was found that high level of NF-kB is highly associated with occurrence of MM. Induction of programmed cell-death (apoptosis) is another target, through which; destruction of tumor cells can be triggered.Aim of the study: to demonstrate the effectiveness of curcumin on expression of NF-kB and induction of apoptosis as compared to the use of arsenic trioxide on B16 cell line.Material and Methods: in this study we utilized B16 cell lines that divided into four categories (curcumin treated, ArT treated, curcumin combined with ArT, and control categories). The first 3 categories where treated with serial concentrations of each drug (at concentrations of 1, 2, 4, 8, 16, and 32 µg/ml), then the supernatant of 96 well of cell culture plate was used for ELISA detection of NF-kB level while cell pellet was used for detection of percentage of apoptotic cells.Results: there was significant reduction of NF-kB level when curcumin was used from 390±2.646 pg/ml to 223.67±11.02 (at P<0.001) while ArT reduced NF-kB from 356+27.683 to 231.67+10.07 pg/ml (at P<0.001). The percentages of apoptotic cells elevated with using of curcumin from 37.4±0.8 to 99.93+0.06% (at P<0.001), while ArT increase in percentages of apoptotic cells from 11.1±0.2% to 99.17+0.17% (at P<0.001). Conclusion: Curcumin had very potent concentration dependent anti-cancer effects against B16 MM cell line. ArT had similar concentration dependent anti-cancer activity but to less extent effect than curcumin against B16 melanoma cell line. Combination of curcumin with ArT had no significant potentiation effect for activity of curcumin on reduction of NF-kB level or increase in the percentage of apoptotic cells.Keywords: curcumin, arsenic trioxide, NF-kB, flow cytometry, apoptotic cell, B16 cell line.