<i>In vitro</i> effects of diclofenac on proliferation and MYC expression in the human melanoma cell line MelIm.

<p>The human melanoma cell line MelIm was incubated with different concentrations of diclofenac (A), aspirin (ASA, B), and NS-398 (C), respectively, and proliferation was determined after 24 h. Results represent the mean +/− standard deviation of 12 (diclofenac) and 3 (ASA, NS-398) independent experiments, respectively. (D) MelIm were incubated for 24 h with or without diclofenac. Apoptotic cells were stained with Annexin-V-FITC/ 7-AAD and analyzed by flow cytometry. Results represent the mean +/− standard deviation of 3 independent experiments. (E-G) MYC, STAT3, HIF1a and HIF2a protein expression were determined in cell lysates of MelIm incubated for 2 or 24 h with or without diclofenac (E,F) or ASA (G). The effect of diclofenac on MYC promoter activity was determined by transient transfection of a 2632-bp MYC promoter fragment (H). MelIm were transfected in 6-well-plates and diclofenac was added after 5 h. Luciferase activity was determined 24 h after transfection. Results represent the mean +/− standard deviation of 3 independent experiments.