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. Author manuscript; available in PMC: 2007 Feb 1.
Published in final edited form as: Cancer Res. 2006 Feb 1;66(3):1712–1720. doi: 10.1158/0008-5472.CAN-05-3138

Fig. 4. FoxM1 is essential for growth and proliferation of PC-3, LNCaP and DU-145 prostate cancer cell lines.

Fig. 4

We transfected siRNA duplexes specific to either FoxM1 (siFoxM1 #2) or p27Kip1 (siP27) into PC-3, LNCaP and DU-145 prostate cancer cell lines and at 48 hours after transfection they were used for growth or flow cytometry analysis and at 72 hours after transfection they were used to prepare total RNA. (A) Transfection of FoxM1 siRNA inhibits expression of FoxM1 in PC-3, LNCaP and DU-145 cells. Total RNA was prepared from prostate cancer cell lines PC-3, LNCaP and DU-145 at 48 hours after transfection with either siRNA duplexes specific to FoxM1 (siFoxM1 #2) or p27Kip1 (siP27) or left untransfected and then was analyzed for expression levels of FoxM1 and Cyclophilin by quantitative Real-Time RT-PCR (QRT-PCR) as described in Material and Methods. FoxM1 mRNA levels in each individual sample were normalized to its corresponding Cyclophilin mRNA level. (B) Transfection of FoxM1 siRNAs into prostate cancer cell lines decreases their growth in culture. Prostate cancer cell lines PC-3, LNCaP and DU-145 were transfected with either siFoxM1 #2 or siP27 duplexes or left untransfected and were then re-plated 48 hours after siRNA transfection and cell numbers were counted at day 3, day 4 or day 5 after transfection. A statistically significant decrease in the growth of FoxM1 depleted prostate cancer cells compared to untransfected cells. (C) Flow cytometry analysis of FoxM1 depleted prostate cancer cell lines shows decreased S-phase progression. The indicated prostate cancer cell lines were transfected with siFoxM1 #2 or siP27 duplexes or left untransfected and then subjected to flow cytometry analysis at 72 hours post transfection after staining with propidium iodide. Graphically shown is the percentage of cells accumulating in G1, S, and G2/M (4N) in FoxM1 or p27Kip1 depleted prostate cancer cells compared to untransfected prostate cancer cells ± SD in triplicate. The asterisks indicate statistically significant increases with P values calculated by Student T Test: *P <0.05, **P ≤ 0.01 and ***P ≤ 0.001.