Profiling of disseminated cells and metastases in MMTV-PyMT hyperplasia transplant model.

Abstract

<p>Representative brightfield (A) and GFP fluorescence (A′) images of the lungs of mice with advanced carcinoma (18 week outgrowths). Disseminated tumors cells were detected as single, GFP-positive cells in the lungs as early as 8-weeks post-transplant. (B) Representative image of a single disseminated tumor cell in the lung of an 18-week-tumor-bearing mouse. Green indicates GFP and blue indicates DAPI. (C) Whole mount image of lung metastases. Mice with 18-week tumor outgrowths were anesthetized and the primary tumor was surgically removed. Mice were sacrificed when they developed the first signs of respiratory distress, indicating the growth of lung metastases (typically 8–10 weeks after primary tumor removal). (D) Number of genes differentially expressed by 2-fold (light blue) and 4-fold (dark blue) in microarray dataset comparisons. (E) Hierarchical clustering of chromatin modifying genes in tumor populations. Tumor populations from adenoma (red), carcinoma (green), disseminated cells (blue), and metastases (purple) were FACS-sorted for profiling. Inset shows clustering of H3K4, H3K9, and H3K27 methyltransferases. Scale bars correspond to 1 mm (A, C) and 5 um (B).</p