Journal: Scientific Reports

Article Title: WNT-1 inducible signaling pathway protein-1 enhances growth and tumorigenesis in human breast cancer

doi: 10.1038/srep08686

Figure Lengend Snippet: (A) Western blot depicting WISP1 expression in MCF7-DNA, MCF7-WISP1-1, and MCF7-WISP1-2 cells. The WISP1 levels of intracellular components (B) and condition media (C) of cells were measured by ELISA. Data are presented as the mean ± SE (n = 6). The effect of ectopic WISP1 expression on MCF-7 cell proliferation was determined by Ki67 expression measured by flow cytometry (D). ISYTYPE represents the negative control group. Data are presented as the mean ± SD (n = 3). 3 H-thymidine incorporation assays (E) were also applied to measure cell proliferation. Data are presented as the mean percentage ± SE (n = 3–6) in relation to mock-transfected (MCF7-DNA) cells. (*P < 0.01).

Article Snippet: MCF7 cells were transduced either with NDRG1 small hairpin RNA lentiviral particles (Sc-36021-V; Santa Cruz Biotechnology) (MCF7-NDRG1si) or with control small hairpin RNA lentiviral particles (Sc-10808-V, Santa Cruz Biotechnology) (MCF7-COLsi) as described by the manufacturer.

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Negative Control, Transfection

Journal: Scientific Reports

Article Title: WNT-1 inducible signaling pathway protein-1 enhances growth and tumorigenesis in human breast cancer

doi: 10.1038/srep08686

Figure Lengend Snippet: (A). The cell cycle distribution of MCF7-DNA and MCF7-WISP1-1 cells was analyzed by flow cytometry after 48 hours incubation. The data shown in each bar chart represent the mean percentage ± SE ( n = 5) of cells in each phase of the cell cycle. (B). The expression (Cropped) and quantitative analysis of cyclins, p21, p27, WISP1, and BTG2 in MCF7-DNA and MCF7-WISP1-1 cells were determined by immunoblotting assays. Data are presented as fold-induction of target genes after WISP1-overexpressed. (* P < 0.01).

Article Snippet: MCF7 cells were transduced either with NDRG1 small hairpin RNA lentiviral particles (Sc-36021-V; Santa Cruz Biotechnology) (MCF7-NDRG1si) or with control small hairpin RNA lentiviral particles (Sc-10808-V, Santa Cruz Biotechnology) (MCF7-COLsi) as described by the manufacturer.

Techniques: Flow Cytometry, Incubation, Expressing, Western Blot

Journal: Scientific Reports

Article Title: WNT-1 inducible signaling pathway protein-1 enhances growth and tumorigenesis in human breast cancer

doi: 10.1038/srep08686

Figure Lengend Snippet: The cell migration (A) and invasion (B) of MCF7-DNA, MCF7-WISP1-1, and MCF7-WISP1-2 cells was determined by trans-well filter without and with Matrigel-coated membranes. The migrating or invading cells were digitally photographed and then counted under the microscope. Experiments were performed in triplicate and repeated at least three times, and the data of quantitative analysis were expressed as average cell counts/9 fields ± SE (*P < 0.01). (C) Gene expression of epithelial-mesenchymal transition markers in MCF7-DNA, MCF7-WISP1-1, and MCF7-WISP1-2 cells was determined by western blot assays (Cropped). The fold-induction data are expressed as the intensity of the protein bands produced by the target gene/β-actin (± SE; n = 3) relative to that of the MCF7-DNA cells (* P < 0.01; + P < 0.05). (D) Immunofluorescence staining of F-actin (red) expression and distribution of MCF7-DNA, MCF7-WISP1-1, and MCF7-WISP1-2 cells. DAPI (blue) was applied for nuclear staining.

Article Snippet: MCF7 cells were transduced either with NDRG1 small hairpin RNA lentiviral particles (Sc-36021-V; Santa Cruz Biotechnology) (MCF7-NDRG1si) or with control small hairpin RNA lentiviral particles (Sc-10808-V, Santa Cruz Biotechnology) (MCF7-COLsi) as described by the manufacturer.

Techniques: Migration, Microscopy, Gene Expression, Western Blot, Produced, Immunofluorescence, Staining, Expressing

Journal: Scientific Reports

Article Title: WNT-1 inducible signaling pathway protein-1 enhances growth and tumorigenesis in human breast cancer

doi: 10.1038/srep08686

Figure Lengend Snippet: NDRG1 expression in MCF7-DNA, MCF7-WISP1-1, and MCF7-WISP1-2 cells was determined by western blot (A) and RT-qPCR (B). NDRG1 expression of MCF-7 cells after treatment with recombinant human WISP1 protein as determined by western blot (top) and RT-qPCR (bottom) (C). (D) The NDRG1 reporter vector containing the human NDRG1 promoter/enhancer DNA fragment (−4714 to +46) was co-transfected with different concentrations of WISP-1 expression vector into MCDF-7 cells. The luciferase activity of the NDRG1 reporter in MCF-7 cells was presented as the mean percentage ± SE (n = 6) in relation to no WISP-1 expression vector transfection group. (E) Relative luciferase activity of reporter vectors containing different fragments from the NDRG1 promoter/enhancer as indicated. Data are presented as mean percentage ± SE (n = 6) of the luciferase activity in relation to mock-transfected cells (* P < 0.01).

Article Snippet: MCF7 cells were transduced either with NDRG1 small hairpin RNA lentiviral particles (Sc-36021-V; Santa Cruz Biotechnology) (MCF7-NDRG1si) or with control small hairpin RNA lentiviral particles (Sc-10808-V, Santa Cruz Biotechnology) (MCF7-COLsi) as described by the manufacturer.

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Recombinant, Plasmid Preparation, Transfection, Luciferase, Activity Assay

Journal: Scientific Reports

Article Title: WNT-1 inducible signaling pathway protein-1 enhances growth and tumorigenesis in human breast cancer

doi: 10.1038/srep08686

Figure Lengend Snippet: (A) The expression of NDRG1 in NDRG1-knockdown MCF-7 (MCF7-NDRG1si) cells and mock-knockdown MCF-7 (MCF7-COLsi) cells were determined by immunoblotting (top) and RT-qPCR (bottom). Data are expressed as mean ± SE ( n = 3) and the NDRG1 level of MCF-COLsi cells is set as 1(*P < 0.01). (B) Cell proliferation of MCF7-COLsi (•) and MCF7-NDRG1si (○) cells was determined by the 3 H-thymidine incorporation assay. Each point on the curve represents the mean percentage ± SE ( n = 4) in relation to Day 1. (C) The cell invasion ability of MCF7-COLsi and MCF7-NDRG1si cells after 48 hours incubation was measured using the matrigel-invasion assay. Experiments were performed in triplicate and repeated at least three times. Data of quantitative analysis are expressed as average cell counts/9 fields ± SE of MCF7-COLsi and MCF7-NDRG1si cells. (* P < 0.01).

Article Snippet: MCF7 cells were transduced either with NDRG1 small hairpin RNA lentiviral particles (Sc-36021-V; Santa Cruz Biotechnology) (MCF7-NDRG1si) or with control small hairpin RNA lentiviral particles (Sc-10808-V, Santa Cruz Biotechnology) (MCF7-COLsi) as described by the manufacturer.

Techniques: Expressing, Knockdown, Western Blot, Quantitative RT-PCR, Thymidine Incorporation Assay, Incubation, Invasion Assay

Journal: Scientific Reports

Article Title: WNT-1 inducible signaling pathway protein-1 enhances growth and tumorigenesis in human breast cancer

doi: 10.1038/srep08686

Figure Lengend Snippet: MCF7-DNA (A) and MCF7-WISP1-1 (B) cells (5 × 10 6 ) were equally mixed with matrigel and then injected subcutaneously into the back area of each nude mouse. The tumor volumes (mm 3 ) in the MCF7-DNA group (•) and the MCF7-WISP1-1 group (○) were measured regularly (C). Data are presented as the mean ± SE. Scale bar, 10 mm. (D) Whole-cell lysates of randomly selected tumor samples from MCF7-DNA (A) and MCF7-WISP1-1 groups were subjected to RT-qPCR. Data are presented as mean fold induction of the WISP1 mRNA (± SE; n = 3) relative to the mock-transfected xenograft groups. ( + P < 0.05, * P < 0.01).

Article Snippet: MCF7 cells were transduced either with NDRG1 small hairpin RNA lentiviral particles (Sc-36021-V; Santa Cruz Biotechnology) (MCF7-NDRG1si) or with control small hairpin RNA lentiviral particles (Sc-10808-V, Santa Cruz Biotechnology) (MCF7-COLsi) as described by the manufacturer.

Techniques: Injection, Quantitative RT-PCR, Transfection

Journal: Scientific Reports

Article Title: WNT-1 inducible signaling pathway protein-1 enhances growth and tumorigenesis in human breast cancer

doi: 10.1038/srep08686

Figure Lengend Snippet: NDRG1 expression in transiently NDRG1-transfected MCF7 (MCF7-NDRG1) (A) and MDA-MB-231 (MDA-NDRG1) (D) cells was determined by western blot and RT-qPCR. Recombinant WISP1 was used to treat MCF-7 and MCF7-NDRG1 cells (B) or MDA-MB-231 and MDA-NDRG1cells (E) with the indicated concentrations for 2 days. Cell proliferation was measured using the CyQUANT cell proliferation assay kit. The cell invasion ability of MCF-7 and MCF7-NDRG1 cells (C) or MDA-MB-231and MDA-NDRG1 cells (F) after 48 hours incubation with 1000 ng/ml of recombinant WISP was measured using the matrigel-invasion assay. Each point of the curve represents the mean percentage ± SE (n = 6) in relation to that of control-solvent cells. ( + P < 0.05, * P < 0.01).

Article Snippet: MCF7 cells were transduced either with NDRG1 small hairpin RNA lentiviral particles (Sc-36021-V; Santa Cruz Biotechnology) (MCF7-NDRG1si) or with control small hairpin RNA lentiviral particles (Sc-10808-V, Santa Cruz Biotechnology) (MCF7-COLsi) as described by the manufacturer.

Techniques: Expressing, Transfection, Western Blot, Quantitative RT-PCR, Recombinant, CyQUANT Assay, Proliferation Assay, Incubation, Invasion Assay, Control, Solvent