Modeling Metastatic Lung and Breast Cancer

Parental (that is, scrambled shRNA) and FGFR1-manipulated D2.A1 cells were injected into the lateral tail veins of 5-week-old female BALB/C mice (The Jackson Laboratory, Bar Harbor, ME, USA). Where indicated, mice bearing D2.A1 pulmonary tumors were treated daily with BGJ-398 (ChemieTek, Indianapolis, IN, USA) or PF-573271 (PF271; Pfizer Pharmaceuticals, New York, NY, USA) at 50 mg/kg by oral gavage. Alternatively, Cdh1 reporter 4T1 cells (1 × 10⁴ cells) were engrafted onto the mammary fat pads of 4-week-old BALB/c mice. Circulating 4T1 tumor cells were isolated from the inferior vena cava at the time of necropsy using 3% sodium citrate. Following lysis of red blood cells, circulating tumor cells were selected for with 5 μg/ml Zeocin (the selectable marker for firefly luciferase). Luciferase-expressing NME cells (1 to 2 × 10⁶ cells) were engrafted onto the mammary fat pads of 5-week-old female nu/nu mice. All bioluminescent images were captured using a Xenogen IVIS 200 preclinical imaging system (Caliper Life Sciences/PerkinElmer, Hopkinton, MA, USA) within the Small Animal Imaging Resource Center at the Case Comprehensive Cancer Center as previously described [5 (link),21 (link),23 (link)].