AC3® exerts cytotoxic and anti-migratory activity and modulates the gene expression of TNF-α, inflammatory mediators, and components of the CD39/CD73/adenosine axis in cutaneous melanoma cell lines.

Cutaneous melanoma (CM) poses a critical public health concern due to its high recurrence rates and resistance to standard treatments. This study investigated the antineoplastic potential of Curcuma Longa L. extract enriched with Bisdemethoxycurcumin (AC3®) on human melanoma cell lines A375 and SK-MEL-28. The A375 and SK-MEL-28 cell lines were cultured according to the manufacturer's recommendations and were subsequently treated with different concentrations of AC3®. The MTT test and fluorescence microscopy were used to assess cell viability. The wound-healing assay was used to assess cell migration following compound treatment. To determine reactive oxygen species (ROS) levels, we used the fluorescence assay with 2,7-dichlorodihydrofluorescein diacetate. Subsequently, the molecular pathways associated with apoptosis were evaluated using RT-PCR, including modulation of the inflammatory profile and purinergic signaling components. AC3® effectively inhibited cell proliferation and migration in both melanoma cell lines. In A375 cells, the compound reduced ROS levels and significantly modulated apoptotic pathways, including upregulation of caspase-3 expression. Similarly, in SK-MEL-28 cells, increased expression of caspase-8 and caspase-3 indicated the activation of both extrinsic and intrinsic apoptotic mechanisms at the transcriptional level. Additionally, AC3® influenced the inflammatory response by increasing the expression of Tumor Necrosis Factor Alpha (TNF-α), Interleukin 6 (IL-6), and NLR Family Pyrin Domain Containing 3 (NLRP3) in A375 cells while downregulating IL-6 and NLRP3 expression in SK-MEL-28 cells, highlighting its effect on processes related to tumor development and progression. Furthermore, the compound modulates the expression and enzymatic activity of ectonucleoside triphosphate diphosphohydrolase 1 (CD39) and ecto-5'-nucleotidase (CD73), key regulators of purinergic signaling, suggesting its potential to affect immune evasion mechanisms in melanoma. These findings position the AC3® compound as promising for cutaneous melanoma by targeting critical pathways involved in tumor progression and cell survival.