Fig. 1.

Silencing of HMMR suppresses cell growth by dephosphorylating Rb in prostate cancer cells. (A) Human prostate cancer PC-3 cells were transfected with control siRNA (si-Cont) or various doses of si-HMMR(#1) for 6 h, and further cultured for 48 h (A, left panel) or 4 days (A, middle panel). Separately, cells were transfected with si-HMMR(#2) or si-HMMR(#3) (A, right panel). Cells were then either observed under microscope or subjected to crystal violet staining assay at 550 nm (****p<0.0001 vs. control). (B, C) Three siRNAs against HMMR were introduced at various doses and cells were lysed at 24 h or 48 h after transfection, followed by western blot analyses. (D) PC3 cells stably overexpressing pCMV6-Flag-HMMR (PC3+HMMR) and control cells (PC3+pCMV6) were cultured for 7 days, and the rate of cell growth was determined by crystal violet staining assays. Separately, twenty to 40 μg of protein was analyzed by western blotting to measure the protein level of flag-HMMR and pRb. Data shown are the mean ± SD of triplicate determinations and representative of two or three independent experiments (*p<0.05 vs. control).