Figure 5. Overexpression of CXCL12 or PGK1 in NSFs facilitates PCa growth in vivo.

(A) PC3^luc cells were implanted alone or mixed with NSF^CXCL12, NSF^PGK1, NSF^Control or CAF^Control cells at a ratio of 1:1 cells into the backs of NOD / SCID mice. At 4 weeks, imaging was performed by Xenogen IVIS imaging system. Luciferase signals from the representative mice are shown.

(B) Quantification of tumor burdens identified by bioluminescence imaging. Increasing tumor size was observed after co-injection NSF^CXCL12 or NSF^PGK1 compared with NSF^Control cells. Co-implantation of CAF^Control cells served as a positive control. * Significant difference from control at p < 0.05. # Significant difference from presence of CAF^Control or PC3 alone at p < 0.05.

(C) Immunofluorensence staining for CAF markers α-SMA and Vimentin in tumor stroma from NSF^Control, NSF^CXCL12, and NSF^PGK1 mixed with PC3^luc. Nuclei were identified by DAPI. All images were captured on Zeiss LSM 510 meta confocal laser scanning microscope. Original magnification 60×, where the bars represent 30 μM.

(D) Quantitative evaluation of the expression of PGK1 in human PCa. Immunostaining intensity was scored as either absent (1), weak (2), moderate (3), or strong (4). The mean expression scores multiplied by % positive cells in the field (Quick’s combined score system) for all benign, PIN, localized PCa are presented in graphical format using error bars with 95% confidence intervals (CI). Statistically significant differences were noted between benign hyperplasia (BPH) and PIN, or localize cancer for P < 0.05 (*).

(E) ELISA analysis of PGK1 level in serum of normal controls and PCa patients. The data are presented as mean ± std. dev. for triplicate determinations and normalized against total protein. *Denotes significant difference from respective controls (p<0.05, ANOVA). n=3 in groups.