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Natural Berry Extracts as a Treatment for Hemangiomas
Gayle M. Gordillo, MD, Ayan Biswas, PhD, Emma C. Clark, BA.
The Ohio State University, Columbus, OH, USA.

Purpose: Current therapies for infantile hemangiomas (IH) have high risk side effect profiles. A proprietary blend of natural berry extracts (NBE) was studied as a possible alternative. Using a validated murine model we have shown that tumor forming endothelial (EOMA) cells escape cell death through induction of multidrug resistance protein 1 (MRP1) to efflux oxidized glutathione out of the nucleus. We sought to determine whether NBE could inhibit MRP1 to induce EOMA cell death and promote tumor involution.
Methods: Calcein exclusion was used to measure MRP1 activity in NBE treated (50 ug/ml) EOMA cells. Oxidized and reduced glutathione were measured in subcellular fractions of NBE treated EOMA cells. NBE induced EOMA cell death was measured in vitro and in vivo using immunocytochemistry for activated Bax, Caspase 9 and cleaved Caspase 3. Mice were given a subcutaneous injection of NBE treated EOMA cells (5 x 106 cell/100 ul) to examine effects on tumor growth in vivo.
Results: NBE treatment significantly inhibited MRP1 activity and nuclear localization. This resulted in accumulation of oxidized glutathione in the nucleus with activation of the intrinsic apoptotic pathway as shown by significantly elevated Bax, Caspase 9 and cleaved Caspase 3. Mice injected with NBE treated EOMA cells developed smaller tumors in vivo with increased intrinsic apoptosis.
Conclusions: This is the first report that NBE inhibits MRP1 resulting in decreased tumor size. MRP1 is overexpressed in many different types of tumors. Thus, NBE may have applications for treatment of IH and many other tumors.


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