The aim of this study is based on the evaluation of anticancer, anti‐inflammatory activities, and cellular uptake of hyaluronic acid nanohydrogel of quercetin tested alone and in combination to a macrolide derivative of rapamycin RAD001 (everolimus) on hormone‐responsive breast cancer cell line MCF‐7. Biological investigations were focused on the receptor mediated cellular internalization of the nanohydrogel and its abilities to reduce secretion of several cytokines (IL‐8, IL‐6, IL‐19), VEGF, and metalloproteases (MMP‐2, MMP‐9) under pro‐inflammatory conditions. Nanohydrogel show a CD44 dependent endocytosis with evident time dependent cytoplasmatic accumulation with abilities to reduce secretion of all cytokines of ∼60% compared to untreated cells. Combination of formulated quercetin and everolimus leads to a synergistic cytotoxic effects with a Combination Index of 0.38. These results highlights the importance of synergistic effect of the hyaluronic acid nanohydrogel of quercetin with everolimus in the regulation of human breast cancer cell proliferation and emphasize the antitumor and anti‐inflammatory properties of the nanocarrier. J. Cell. Physiol. 232: 2063–2074, 2017. © 2016 Wiley Periodicals, Inc. Hyaluronic acid nanohydrogel of Quercetin has anti‐anticancer and antinflammatory activity in human breast cancer cells. Combination of Everolimus and hyaluronic acid nanohydrogel of Quercetin leads to synergistic cytotoxicity in MCF‐7 call line.