miR-200b is sufficient to decrease tumor growth
A) Forced expression of miR-200b in PC3 cells. PC3 cells were trasduced with a bicistronic lentiviral shuttle vector (pMIRNA1 pCDH-CMV-MCS-EF1-copRFP) encoding hsa-miR-200b and empty vector control (ctrl). Total RNA was isolated and miR- 200b expression measured using real-time RT-PCR. The values represent three independent experiments performed in triplicate. *, p≤0.01. (B) miR-200b reduced tumorigenesis by PC-3 cells. Parental PC3 cells, PC3 cells trasduced with miR control (ctrl) and miR-200b were subcutaneously injected into the rear hindquarters of athymic male mice (n = 5). Tumor weight at day 29 post-injection is shown. *, p≤0.05; **, p≤0.01. (C) The tumors maintained miR-200b expression. Relative miR-200b expression was measured by real-time RT-PCR in the tumors from panel (B). * p≤0.05. (D). miR 200b decreased tumor growth in an orthotopic model of prostate cancer. RFP-tagged PC3-ctrl and PC3-200b cells were implaned in the prostates of athymic male mice. The average fluorescence was measured 20 days post injection. (E) Fluorescence per animal was determined using whole body imaging, with Olympus OV100 system. *, p≤0.05. (F) Cell growth was measured by WST-1 assay. PC3-ctrl or PC3 miR-200b cells were seeded at 3000 cells per well in a 96-well plate. Absorbance was measured at indicated time points using a Biorad Model 680 microplate reader. The results represent the average of three independent experiments performed in triplicate. *, p≤0.05 by Student’s T-test. (G, H) Tumor sections were stained for Ki-67, to evaluate proliferation. Note a significant decrease in Ki-67–positive nuclei in the presence of miR-200b. *, p<0.001.
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Williams LV, Veliceasa D, Vinokour E, Volpert OV. miR-200b Inhibits Prostate Cancer EMT, Growth and Metastasis. (2013) PLoS One 8(12):e83991. [article]