Copper Tolfenamic acid induces anti-proliferative activity effective against Medulloblastoma cells

Purpose: Medulloblastoma (MB) is the most common pediatric malignant brain tumor, comprising 20% of all childhood brain tumors. Between 250-500 children per year are diagnosed in the US alone. Standard therapies result in severe long-term morbidities. Therefore, there is an urgent need for inventing novel effective treatment strategies with lower side-effects. Our laboratory showed anti-cancer activity of Tolfenamic acid (TA) in pre-clinical model for MB. Recent studies showed higher pharmacological effect of TA when synthesized as a complex with copper (Copper-TA, Cu-TA). Our aim was to investigate the anti-cancer activity of Cu-TA against MB cell lines. We hypothesize that Cu-TA presents higher anti-cancer activity and is more effective than TA to induce cytotoxicity against MB cells. Methods: DAOY and D283 cells were obtained from ATCC and grown following standard cell culture conditions. Cells were treated with TA or Cu-TA and the cell viability was measured at 24 and 48 h post-treatment using a CellTiter-Glo kit. The induction of apoptosis was investigated by studying caspase activation using the Caspase 3/7-Glo kit. In addition, reactive oxygen species (ROS) involvement was measured by flow cytometry. Results: Both Cu-TA and TA treatment resulted in decreased cell viability. However, when compared to TA, Cu-TA was more effective at inducing anti-proliferative activity in MB cells. Cu-TA induces increased production of ROS. The anti-proliferative activity of Cu-TA was accompanied by an increase in Caspase 3/7 activity, suggesting the induction of apoptosis.

Conclusions: Cu-TA was more effective than TA. Therefore, it has potential as an effective anti-cancer agent for inhibiting MB cell growth. Further studies are needed to better understand Cu-TA’s mechanism of action.