Fig. 8
Effect of therapeutic agents on the growth of TNBC PDX in vivo and ex vivo. (A) BC-AR485 growth curves: Tumors were transplanted by trocar into NSG female mice (n = 4/group). When the tumors reached a size of ~ 25 mm2, fulvestrant (FULVE; 250 mg/kg/week s.c.), nab-paclitaxel (Nab-PAX; 15 mg/kg/4 days per week i.v.), and pegylated liposomal doxorubicin (DOXO-L; 4.5 mg/kg/week i.v.) treatments were initiated and continued for 18 days. Tumor size was measured every 2–3 days and plotted. (B) Ki67 expression. At the end of the experiment, tumors were excised, processed for IHC studies and the expression of Ki67 was evaluated. The X̅ ± SD of 10 randomly fields were quantified and a representative image is shown. Black arrow shows an extensive area of necrosis (DOXO-L) and macronuclei induced treatment (Nab-PAX). (C) BC-AR687 growth curves. Tumors were transplanted by trocar into NSG female mice (n = 4/group). When the tumors reached a size of ~ 25 mm2, mice were treated with everolimus (EVERO-a, Afinitor Novartis) 5 mg/kg/5days a week p.o., alpelisib (ALPE-p, Piqray Novartis), 30mg/kg/5 days a week per os (p.o.) and rapamycin (RAPA, MCE) 17.5mg/kg/2 days a week intraperitoneally (i.p.). (D) Ki67 expression. The X̅ ± SD of 10 randomly fields were quantified and a representative image is shown. (E) Scheme of breast cancer tissue cultures (BCTC) from PDX. Tumors were excised, cut in thin slices with a Vibratome and pieces were cultured for 48 hs with therapeutic agents. Then tissues were formalin-fixed and embedded in paraffin to quantify pS6 and KI67 expression evaluated by immunohistochemistry (IHC). (F) The X̅ ± SD of 10 randomly fields were quantified and representative images are shown. Top: Ki67 index in control, DOXO- or PAX-treated cultures. Bottom: pS6 index in alpelisib (ALPE)- or everolimus (EVERO)-treated cultures. *: p < 0.05; **: p < 0.01; ***: p < 0.001; ****: p < 0.0001. Bar: 100 µM.
