Journal: International journal of oncology

Article Title: MicroRNA-20b promotes cell proliferation via targeting of TGF-β receptor II and upregulates MYC expression in Ewing's sarcoma cells.

doi: 10.3892/ijo.2017.4155

Figure Lengend Snippet: Figure 1. Whole genome array analysis in ES cell lines. (A) miRNA expression in five ES cell lines (SCCH, RDES, WE68, SK-ES-1 and SKNMC) and hMSCs. Heat maps of mRNA expression and extraction of TGFBR2 (B) and MYC (C) in ES cells and hMSCs. The color bar shows the relative expression levels; red and blue indicate an increase and decrease, respectively.

Article Snippet: The TGFBR2 expression plasmid (SC119965) was obtained from Origene Technologies Inc. (Rockville, MD, USA).

Techniques: Expressing, Extraction

Journal: International journal of oncology

Article Title: MicroRNA-20b promotes cell proliferation via targeting of TGF-β receptor II and upregulates MYC expression in Ewing's sarcoma cells.

doi: 10.3892/ijo.2017.4155

Figure Lengend Snippet: Figure 2. Inhibition by miR-20b of TGFBR2 mRNA expression and repressing of TGFBR2 with anti-miR-20b in SKES1 cells. (A) Possible binding sites of miR-20b at the 3'-UTR site of TGFBR2 mRNA. Each sequence of miR-20b (Wt) and its mutant (Mut). (B) After actinomycin D administration, the miR-20b (B) and mRNA (C) expression levels (B) in the negative control-miR, miR-20b and miR-20b mutant were evaluated by qRT-PCR. (D) Western blot analysis showed increased TGFBR2 protein in anti-miR-20b and TGFBR2-expression vector group. (E) Densitometry evaluation of TGFBR2 protein. (F) Western blot analysis showing increased expression of TGFBR2 protein following transfection of TGFBR2 expression vector in SKES1 cells. (G) Expression of TGFBR2 protein after induction of TGFBR2 vector. **p<0.01.

Article Snippet: The TGFBR2 expression plasmid (SC119965) was obtained from Origene Technologies Inc. (Rockville, MD, USA).

Techniques: Inhibition, Expressing, Binding Assay, Sequencing, Mutagenesis, Negative Control, Quantitative RT-PCR, Western Blot, Plasmid Preparation, Transfection

Journal: International journal of oncology

Article Title: MicroRNA-20b promotes cell proliferation via targeting of TGF-β receptor II and upregulates MYC expression in Ewing's sarcoma cells.

doi: 10.3892/ijo.2017.4155

Figure Lengend Snippet: Figure 3. Cell growth assay of the effects of anti-miR-20b. Anti-miR-20b inhibits cell growth of SK-ES-1 cells (A) and RDES cells (B). TGFBR2 expression vector inhibits cell growth of SK-ES-1 cells (C) and RDES cells (D). Data represent the mean ± SD of three independent experiments. p<0.05 was considered to indicate significance: *p<0.05, **p<0.01.

Article Snippet: The TGFBR2 expression plasmid (SC119965) was obtained from Origene Technologies Inc. (Rockville, MD, USA).

Techniques: Growth Assay, Expressing, Plasmid Preparation

Journal: International journal of oncology

Article Title: MicroRNA-20b promotes cell proliferation via targeting of TGF-β receptor II and upregulates MYC expression in Ewing's sarcoma cells.

doi: 10.3892/ijo.2017.4155

Figure Lengend Snippet: Figure 4. Immunoblot analysis of TGFBR2, MYC and TGF-β signaling-related factors. Effects of anti-miR-20b and TGFBR2 vector on TGFBR2, MYC and TGF-β pathway in (A) SK-ES-1 cells and (B) RDES. (C) The quantification of western blot analysis. Data represent the mean ± SD of three independent experiments. p<0.05 was considered to indicate significance: *p<0.05, **p<0.01.

Article Snippet: The TGFBR2 expression plasmid (SC119965) was obtained from Origene Technologies Inc. (Rockville, MD, USA).

Techniques: Western Blot, Plasmid Preparation

Journal: International journal of oncology

Article Title: MicroRNA-20b promotes cell proliferation via targeting of TGF-β receptor II and upregulates MYC expression in Ewing's sarcoma cells.

doi: 10.3892/ijo.2017.4155

Figure Lengend Snippet: Figure 5. Anti-miR-20b and TGFBR2 expression vector treatment induces cell cycle retardation. Cells were untreated (A) or transfected with negative control miR (B), anti-miR-20b (C) and TGFBR2 expression vector (D), stained with propidium iodide and analyzed for cell cycle distribution. Histogram shows the mean values of each cell cycle phase (E): *p<0.05, **p<0.01. Detection of cleaved PARP levels present in protein levels by immunoblot. Anti-miR-20b and TGFBR2 expression vector with low-dose adriamycin treatment induces apoptosis (F). The quantification of PARP and cleaved PARP protein (G). The above experimental results were analyzed for the presence of Annexin V (+)/PI (-) (early apoptosis) and Annexin V (+)/PI (+) (late apoptosis). SKES1 cells treated with anti-miR-20b and TGFBR2 expression vector and anti-miR-20b and TGFBR2 expression vector with low-dose adriamycin were stained with Annexin V-FITC/PI, and analyzed for cell apoptosis distribution (H).

Article Snippet: The TGFBR2 expression plasmid (SC119965) was obtained from Origene Technologies Inc. (Rockville, MD, USA).

Techniques: Expressing, Plasmid Preparation, Transfection, Negative Control, Staining, Western Blot

Journal: International journal of oncology

Article Title: MicroRNA-20b promotes cell proliferation via targeting of TGF-β receptor II and upregulates MYC expression in Ewing's sarcoma cells.

doi: 10.3892/ijo.2017.4155

Figure Lengend Snippet: Figure 7. A flowchart miR-20b axis in the attenuation of TGFBR2 signaling in Ewing's sarcoma cell lines.

Article Snippet: The TGFBR2 expression plasmid (SC119965) was obtained from Origene Technologies Inc. (Rockville, MD, USA).

Techniques:

Journal: International journal of oncology

Article Title: MicroRNA-20b promotes cell proliferation via targeting of TGF-β receptor II and upregulates MYC expression in Ewing's sarcoma cells.

doi: 10.3892/ijo.2017.4155

Figure Lengend Snippet: Figure 6. Suppression of ES tumor growth by anti-miR-20b and TGFBR2 vector in vivo. (A) Four groups of mice inoculated with: i) untreated SK-ES-1 cells (n=7); ii) transfected with negative control-miR (n=7); iii) transfected with anti-miR-20b (n=7); and iv) transfected with TGFBR2 expression vector (n=7). Tumor volumes were measured every 3 days during treatment. **p<0.01. (B) Immunohistochemistry image analysis of TGFBR2 and MYC following transfec- tion of anti-miR-20b and TGFBR2 vector. (C) Number of TGFBR2- and MYC-positive cells per unit area.

Article Snippet: The TGFBR2 expression plasmid (SC119965) was obtained from Origene Technologies Inc. (Rockville, MD, USA).

Techniques: Plasmid Preparation, In Vivo, Transfection, Negative Control, Expressing, Immunohistochemistry