Evaluation of a Synergistic Effect of L-Arginine on the Anticancer Activity of Doxorubicin by Using a Co-culture System

In the early stage of tumor development, tumor-associated macrophages (TAM) works to suppress tumor growth by secreting soluble factors including nitric oxide (NO). L-Arginine (Arg) is a substrate of nitric oxide synthase (NOS) expressed in TAM. Here we examined whether NO produced from Arg by macrophages works to enhance the effect of the anti-cancer drug, doxorubicin (Dox) by using a co-culture system of cancer cells with macrophages. By employing colorimetric analyses methods (Griess Reagent and Cell Counting kit-8), we found that NO produced from Arg by co-cultured macrophages could enhance the cytotoxic effect of Dox to cancer cells. Moreover, we found that augmentation is affected by the order of the addition of Arg and Dox. A prior addition of Arg to Dox and simultaneous addition showed the same enhancement effect, but a prior addition of Dox to Arg abolished the augmentation. This suggests that the co-administration of Arg with Dox would be an effective treatment to improve chemo-therapies.