FIGURE 3.

Chemokine receptor CXCR4 is functionally essential target of wt-p53 in prostate cancer bone metastasis. (A, B) Western blot analysis of CXCR4 expression in the indicated cells. α-Tubulin served as a loading control. (C) Wound-healing assays for the indicated cell lines. Wound closure was photographed at 24 h after wounding. Tumor cells were cultured in MC3T3-E1 condition medium. (D) Quantification of the migration of indicated cells in transwell assays. Tumor cells (top chamber) were co-cultured with primary osteoblasts (bottom chamber) in the transwell plate. (E) Quantification of in vitro competitive colocalization assays for the indicated cells. Error bars represent the means ± SD. (F) Representative BLI images of bone lesions from mice intracardially inoculated with prostate cancer cells in each experimental group (n = 8/group). (G) Histomorphometric quantification of tumor area in hind limbs from each experimental group. (H) Numbers of metastatic lesions in hind limb bones from each mouse in two groups. Error bars represent the means ± SEM. (I) Kaplan-Meier curves of bone metastasis-free survival (BMFS) and overall survival (OS) in mice from each experimental group (log-rank test). P values were based on t-test unless otherwise indicated. *p < 0.05, **p < 0.01, ***p < 0.001.