Figure 5
ANO1-dependent stimulation of prostate cancer cell bone metastasis by 5-Aza-CdR. (A) Xenografts were established by injecting mock-expressing control or ANO1-expressing LNCap and PC3 cells into the distal femur and proximal tibia of mice and treated with DMSO or 5-Aza-CdR for 30 days. Tumor growth was monitored in LNCap and PC3 xenografts by detecting bioluminescence with an in vivo imaging system (IVIS) after 1 and 30 days of treatments. The bioluminescence intensity was quantified using regions of interest (ROIs) of equivalent-sized areas at the indicated time. Data represent the mean ± SD (n = 5); ***P < 0.001 versus LNCap-luc. (B) After treating mice bearing LNCap and PC3 xenograft as in (A), mice were sacrificed, and the bones of the hindlimb were excised from the distal legs. The bone metastatic tumor cells on the hindlimb were visualized by detecting bioluminescence with an IVIS. (C) Total RNA samples were isolated from tibial bone in mice for 30 days after DMSO or 5-Aza-CdR treatment. RT-qPCR was performed using primers listed in Supplementary Table S1. All transcription levels were normalized to that of GAPDH. Data represent the mean ± SD of three independent experiments in triplicate; ***P < 0.001 versus LNCap-luc.
