Prostate Tumor Xenograft Model

Approximately 4-week-old male NOD.CB17-Prkdcscid/J mice were purchased from the Jackson Laboratory (Bar Harbor, MI, USA). All animal procedures were verified and approved by the Institutional Animal care and Use Committee of UTMDACC. To study primary prostate tumor formation, 2 × 10⁶ RFP–luciferase-labeled DU145 cells were injected s.c. on both the flanks of 6-week-old male NOD/SCID mice. Once palpable tumors were discernable, tumor-bearing mice were randomly segregated into four groups, and drug treatment was initiated every 24 h for 5 days per week. SB203580 (0.2 μmol in 100 μl per ~20 g mouse), Enzalutamide (10 mg/kg) or a combination of both drugs (or vehicle) were administered s.c., and tumor growth was assessed as described previously.^{17 (link)} Investigators were blinded to the group allocation while assessing experimental outcomes. At the end of the treatment period, tumors were excised, average diameter calculated using calipers and tumor weight noted. Tumors were then processed for RNA isolation and/or fixed in formalin, paraffin-embedded, sectioned and stained with hematoxylin/eosin and pATF2, AR and FOXC2 antibodies.