Figure 2
CCL2-mediated progression of patient derived breast xenografts is associated with increased CCR2, phospho-SMAD3 and phospho-p42/44MAPK expression. N = 4 mice (8 mammary fat pads total) were injected each with DCIS cells from patients IC-041717-1 and IC-031317. N = 2 mice (4 mammary fats total) were injected with DCIS cells from patient IC-022316. Mice were treated with placebo or recombinant CCL2 for up to 22 weeks. Primary breast xenografts were co-immunofluoresence stained for CK5/CK19 (red) with (A) α-sma, (B) PCNA, (C) CCR2, (D) phospho-SMAD3 (green) or E. phospho-p42/44MAPK (red). Sections were counter stained with DAPI. Representative images are shown for case no. IC-031317 with CK5/CK19 overlay with α-sma, PCNA or CCR2, and DAPI overlay with phospho-SMAD3 or phospho-p42/44MAPK expression. Blue arrows point to invasive carcinoma cells, characterized by absence of α-sma staining and presence of cancer cells in the peri-ductal stroma. White arrows point to positive PCNA staining. Scale bar = 200 microns. Sample size of mammary lesions per group are shown below the last graph. Invasiveness was scored for lesions co-stained for α-sma/CK5/19. Expression was quantified by Image J. Stacked bar graph is shown is shown for percentage of invasive and non-invasive lesions (A). Whisker box plots are shown for (B–E). Whiskers indicate min and max values. Box indicates upper and lower quartile range. Line indicates median. Statistical analysis was performed using Chi Square Test (A) or Two tailed T-test (B–E). Statistical significance analysis was determined by p < 0.05. ns not significant.
