Journal: PLoS ONE

Article Title: High Expression of CD109 Antigen Regulates the Phenotype of Cancer Stem-Like Cells/Cancer-Initiating Cells in the Novel Epithelioid Sarcoma Cell Line ESX and Is Related to Poor Prognosis of Soft Tissue Sarcoma

doi: 10.1371/journal.pone.0084187

Figure Lengend Snippet: A. Expression of CD109 mRNA in ES cell lines. Bars represent mean±SEM. ※※※ p <0.001, determined by the Mann-Whitney test. N.S.: not significant. B. Expression of CD109 mRNA in human sarcoma cell lines. Cell lines of epithelioid sarcoma (FU-EPS-1 and VA-ES-BJ), osteosarcoma (OS2000, KIKU, NY, U2OS, Saos-2, HuO9 and HOS), Ewing sarcoma (SKES, WES and RDES), synovial sarcoma (Fuji and YaFuSS) and malignant fibrous histiocytoma (MFH2003 and MFH2004) were used. C. Expression of CD109 mRNA in human fetal tissues (upper panel) and human adult tissues (lower panel). ESX was used as a positive control. D. Immunohistochemistry of CD109 in normal adult tissues.

Article Snippet: Human osteosarcoma cell lines (NY, U2OS and HOS), human Ewing sarcoma cell lines (SKES, WES, and RDES), the human synovial sarcoma cell line FUJI, and the human ES cell line VA-ES-BJ were purchased from the Japanese Collection of Research Bioresources Cell Bank (Tokyo, Japan) and American Type Culture Collection (Manassas, VA, USA).

Techniques: Expressing, MANN-WHITNEY, Positive Control, Immunohistochemistry

Journal: PLoS ONE

Article Title: c-Myc Represses Tumor-Suppressive microRNAs, let-7a, miR-16 and miR-29b, and Induces Cyclin D2-Mediated Cell Proliferation in Ewing’s Sarcoma Cell Line

doi: 10.1371/journal.pone.0138560

Figure Lengend Snippet: (A), Transfection of c-Myc siRNA in SKES1 reduces the expression of c-Myc protein. (B), The quantification of c-Myc protein after transfection of c-Myc siRNA. (C), miRNA quantification of let-7a, miR-16 and miR-29b expression after transfection with c-Myc siRNA by quantitative RT-PCR. (D-F,) After actinomycinD treatment, the miRNA expression level of let-7a, miR-16 and miR-29b in the negative control-miR, let-7a, miR-16, miR-29b and their mutant was measured by qRT-PCR. After actinomycinD treatment, the mRNA expression level of CCND2 after transfection of negative control-miR, let-7a, miR-16, miR-29b and their mutant was measured by qRT-PCR. ANOVA was performed to statistically analyze the data. * p < 0.05 and ** p < 0.01.

Article Snippet: The human ES cell lines, SKES1, RDES, SKNMC and SCCH were obtained from JCRB Cell Bank (Tokyo, Japan), and WE68 was kindly provided Dr. Frans van Valen (Westfalische-Wilhelms University, Munster, Germany).

Techniques: Transfection, Expressing, Quantitative RT-PCR, Negative Control, Mutagenesis

Journal: PLoS ONE

Article Title: c-Myc Represses Tumor-Suppressive microRNAs, let-7a, miR-16 and miR-29b, and Induces Cyclin D2-Mediated Cell Proliferation in Ewing’s Sarcoma Cell Line

doi: 10.1371/journal.pone.0138560

Figure Lengend Snippet: The expression of CCND2 protein was decreased in SKES1 transfected with let-7a (A), miR-16 (C), miR-29b (E) and CCND2-siRNA (G). Densitometry quantification of CCND2 protein levels after transfection of let-7a (B), miR-16 (D), miR-29b (F) and CCND2-siRNA (H). * p < 0.05 and ** p < 0.01.

Article Snippet: The human ES cell lines, SKES1, RDES, SKNMC and SCCH were obtained from JCRB Cell Bank (Tokyo, Japan), and WE68 was kindly provided Dr. Frans van Valen (Westfalische-Wilhelms University, Munster, Germany).

Techniques: Expressing, Transfection

Journal: PLoS ONE

Article Title: c-Myc Represses Tumor-Suppressive microRNAs, let-7a, miR-16 and miR-29b, and Induces Cyclin D2-Mediated Cell Proliferation in Ewing’s Sarcoma Cell Line

doi: 10.1371/journal.pone.0138560

Figure Lengend Snippet: The SKES1 were treated and analyzed by flow cytometry after PI staining (A–E). Histogram shows the quantitative percentage of diploid cells (DNA content) in each cell-cycle phase (F). Effects of let-7a, miR-16 and miR-29b on the induction of apoptosis in SKES1 cells. Western blotting shows the expression of PARP (G) and its cleaved form (H). The cells were labeled with FITC annexin V and PI (I–N). Each quadrants represent viable cells (Lower Left quadrant), early apoptotic cells (Lower Right), late or secondary necrotic cells (Upper Right), and primary necrotic cells (Upper Left), respectively. * p < 0.05.

Article Snippet: The human ES cell lines, SKES1, RDES, SKNMC and SCCH were obtained from JCRB Cell Bank (Tokyo, Japan), and WE68 was kindly provided Dr. Frans van Valen (Westfalische-Wilhelms University, Munster, Germany).

Techniques: Flow Cytometry, Staining, Western Blot, Expressing, Labeling

Journal: PLoS ONE

Article Title: c-Myc Represses Tumor-Suppressive microRNAs, let-7a, miR-16 and miR-29b, and Induces Cyclin D2-Mediated Cell Proliferation in Ewing’s Sarcoma Cell Line

doi: 10.1371/journal.pone.0138560

Figure Lengend Snippet: (A), Transfection of c-Myc siRNA in SKES1 reduced the expression of CCND2 protein. (B), Transfection of CCND2 siRNA in ES cells reduced the expression of CCND2 protein. The cells were treated and analyzed by flow cytometry after PI staining (C–F). Histogram shows the quantitative percentage of diploid cells (DNA content) in each cell-cycle phase in c-Myc and CCND2-siRNA treated cells (G). Effects of c-Myc and CCND2-siRNA on induction of apoptosis in the SKES1 cells. The cells were labeled with FITC annexin V and PI (H–L). * p < 0.05.

Article Snippet: The human ES cell lines, SKES1, RDES, SKNMC and SCCH were obtained from JCRB Cell Bank (Tokyo, Japan), and WE68 was kindly provided Dr. Frans van Valen (Westfalische-Wilhelms University, Munster, Germany).

Techniques: Transfection, Expressing, Flow Cytometry, Staining, Labeling