Colonies was much fewerwhen RNPC1a was over-expressed (Figure. 3C, pColonies was much fewerwhen RNPC1a was

Colonies was much fewerwhen RNPC1a was over-expressed (Figure. 3C, p
Colonies was much fewerwhen RNPC1a was over-expressed (Figure. 3C, p < 0.05). The ability of MCF-7 or MB-231 cell lines to form colonies was much more when RNPC1a was knockdown (Figure. 3D, p < 0.05).RNPC1a suppressed migratory and invasive potentialAs shown in Figure 4A and C, determined by their migration in the wound gap after 18 h, distance migrated ofXue et al. BMC Cancer 2014, 14:322 http://www.biomedcentral.com/1471-2407/14/Page 8 ofFigure 3 RNPC1a suppressed anchorage dependent growth of breast cancer cells. (A, B) Cell cycle progression was measured using flow cytometry. The progression of MCF-7-RNPC1a and MB-231-RNPC1a cells was arrest in the G1 phase compared with control cells, respectively. Representative photographs (upper) and quantification (lower) are shown. (C) The growth of cells over 15 days was measured using colony formation assays. Clone formation of RNPC1a overexpression arbitrarily set at 100 in control cells (NC). The number and size of MCF-7-RNPC1a or MB-231-RNPC1a was significantly decreased compared to control cells, respectively. Representative photographs (lower) and quantification (upper) are shown. Data were means of three separate experiments mean ?SEM, p < 0.05. (D) Clone formation of RNPC1a knockdown arbitrarily set at 100 in knockdown (shRNPC1a) cells. The number and size of MCF-7-shRNPC1a or MB-231-shRNPC1a was significantly increased compared with control cells, respectively. Representative photographs (lower) and quantification (upper) are shown. Data were means of three separate experiments mean ?SEM, p < 0.05. Colonies > 50 mm PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/25962748 were RDX5791 web counted. Anchorage ependent growth assays were shown at the bottom. Data were means of three separate experiments mean ?SEM, *p < 0.05.RNPC1a overexpression decreased by 69 m (Figure 4A, p < 0.01), while RNPC1a knockdown increased by 110 m (Figure 4C, p < 0.01) compared to the control cells, respectively. We conducted three-dimensional cell migration assay using transwell chambers and invasion assay with Matrigel-precoated transwell chambers. We found that RNPC1a overexpression exhibited significantly decrease ability of migration and invasion (Figure 4B, both p < 0.01). RNPC1a knockdown exhibited significantly increase ability of migration and invasion (Figure 4D, bothp < 0.05). Besides, we obtained the similar results of MCF7 cells (Additional files 3: Figure S3).RNPC1a down-regulate mutp53 and up-regulate p21 protein expression in breast cancer cellsPrevious study affirmed that translational of wild-type p53 (wtp53) was repressed by RNPC1a [17]. However, our study found wtp53 protein was no significantly altered in RNPC1a over-expressed or silent MCF-7 cells (Figure 5A). Level of p21 protein was increased in RNPC1aXue et al. BMC Cancer 2014, 14:322 http://www.biomedcentral.com/1471-2407/14/Page 9 ofFigure 4 RNPC1a significantly decreased migratory and invasive potential of breast cancer cells. (A, C) Wound healing assay. Images of wound repair were taken at 0, 18 h after wound. The distance of wound closure is shown by area at 18 h. Representative photographs (upper) and quantification (lower) are shown, original magnification, ?00. (B, D) Transwell migration assay and Matrigel invasion assay. Representative photographs (upper) and quantification (lower) are shown. Columns: average of three independent experiments, *p < 0.05, **p < 0.01, original magnification, ?00.over-expressed MCF-7 and MDA-MB-231 cells (Figure 5A and B). Mutp53 protein was decreased in RNPC1a.