AS1411 affects NCL target miRNAs and cancer cell proliferation in vivo. (A) MDA-MB-231 cells were injected subcutaneously into the flanks of nude mice (n = 20), and at 1 wk after injection, mice with comparable tumor sizes (n = 16) were selected for treatment. Mice were treated with subcutaneous injection near the tumor area of 10 µM AS1411 or DCT (eight mice for each group of treatment). The graph represents the mean tumor volume (mm3) at the indicated days during the treatments for the two groups. Values represent the mean ± SD (n = 8). *, P < 0.01. (B and C) Immunoblot analysis showing DICER, NCL, VIMENTIN, total AKT, phosphorylated AKT (pAKT), PTEN, and BCL-2 protein expression levels from xenograft tumors of treated mice (AS1411 and DCT). GADPH and β-ACTIN were the internal loading control. (C) Densitometric analysis is reported ± SD. *, P < 0.01. (D) qRT-PCR analysis of mature miR-21, miR-221, miR-222, miR-103, miR-30a, and let-7a on extracted tumoral RNA from AS1411-treated samples. Values represent mean ± SD from three independent sets of experiments performed in triplicate. *, P < 0.05 in comparison with the DCT-treated cells. (A–D) All data reported are representative of at least three independent experiments. (E) Hematoxylin and eosin (H&E) and IHC analysis for Ki-67, vimentin, fibronectin, active Caspase-3, pAKT, and NCL in subcutaneous tumors. (F) Quantitative analysis of cytosolic NCL-positive staining from AS1411-treated mice (n = 8) compared with the control group (n = 8). All results are presented as mean ± SD. *, P < 0.01.
