RANK is an independent biomarker of poor prognosis in estrogen receptor-negative breast cancer and functionally contributes to increase tumor aggressiveness

Analyses of genes expression in estrogen receptor negative, RANK positive, breast cancer orthoxenografts after in vivo treatment (4 weeks) with RANKL , RANK-Fc or denosumab. Overall design: Dissociated breast tumor cells mixed 1:1 with Matrigel Basement Membrane (BD Biosciences) were transplanted orthotopically in the inguinal mammary gland of 10/12-week old NSG mice and when tumors reached 5 mm of diameter mice were randomized for mock, h-RANKL (0.75 mg/kg, 4-6 doses, twice per week; Amgen Inc), h-RANK-Fc treatment (10 mg/kg, three times per week; Amgen Inc), denosumab (10 mg/kg, three times per week; XGEVA®). Mice were sacrificed 24 h after the last treatment and RNA extracted from the tumors.