Journal: International Journal of Oncology

Article Title: Overexpression of KH-type splicing regulatory protein regulates proliferation, migration, and implantation ability of osteosarcoma

doi: 10.3892/ijo.2016.3601

Figure Lengend Snippet: Characteristics of patients with osteosarcoma and non-cancer participants.

Article Snippet: Osteosarcoma cell line (MMNG-HOS) catalog no. CLS 300289 passage 24 and U2OS catalog no. CLS 300364 passage 24 were used as representative osteosarcoma cells.

Techniques: Biomarker Discovery

Journal: International Journal of Oncology

Article Title: Overexpression of KH-type splicing regulatory protein regulates proliferation, migration, and implantation ability of osteosarcoma

doi: 10.3892/ijo.2016.3601

Figure Lengend Snippet: Representative 2D-PAGE of osteosarcoma and osteoblast. (A) Red arrows show spots significantly upregulated in osteosarcomas; green arrows show spots significantly upregulated in osteoblasts. (B) Heat map shows fold changes of % volume of protein spots between osteosarcoma and osteoblast in each matched set.

Article Snippet: Osteosarcoma cell line (MMNG-HOS) catalog no. CLS 300289 passage 24 and U2OS catalog no. CLS 300364 passage 24 were used as representative osteosarcoma cells.

Techniques:

Journal: International Journal of Oncology

Article Title: Overexpression of KH-type splicing regulatory protein regulates proliferation, migration, and implantation ability of osteosarcoma

doi: 10.3892/ijo.2016.3601

Figure Lengend Snippet: Summary of significant altered proteins in primary osteosarcoma cells identified by LC-MS/MS.

Article Snippet: Osteosarcoma cell line (MMNG-HOS) catalog no. CLS 300289 passage 24 and U2OS catalog no. CLS 300364 passage 24 were used as representative osteosarcoma cells.

Techniques:

Journal: International Journal of Oncology

Article Title: Overexpression of KH-type splicing regulatory protein regulates proliferation, migration, and implantation ability of osteosarcoma

doi: 10.3892/ijo.2016.3601

Figure Lengend Snippet: (A) KSRP expression levels in experimental samples (1, pooled sample of osteosarcoma; 2, pool sample of osteoblasts; 3–15 (odd number), individual osteosarcoma cases; 2–16 (even number), individual osteoblast cases; 17, MNNG-HOS; 18, U2OS; 19, HeLa cells as the positive control). (B) KSRP expression in 12 representative biopsy samples of osteosarcoma cases in the separate group.

Article Snippet: Osteosarcoma cell line (MMNG-HOS) catalog no. CLS 300289 passage 24 and U2OS catalog no. CLS 300364 passage 24 were used as representative osteosarcoma cells.

Techniques: Expressing, Positive Control

Journal: International Journal of Oncology

Article Title: Overexpression of KH-type splicing regulatory protein regulates proliferation, migration, and implantation ability of osteosarcoma

doi: 10.3892/ijo.2016.3601

Figure Lengend Snippet: Significantly decreased migratory behavior of osteosarcoma cell lines after KSRP knock-down by siRNA-1 (siR-1) and si-RNA2 (siR-2) compared with non-sense (NS) siRNA as a control (A) at 48 h and (B) at 72 h. (C) Cell growth curve showing significantly decreased proliferative ability of osteosarcoma cell lines after KSRP knock-down.

Article Snippet: Osteosarcoma cell line (MMNG-HOS) catalog no. CLS 300289 passage 24 and U2OS catalog no. CLS 300364 passage 24 were used as representative osteosarcoma cells.

Techniques: Knockdown, Control

Journal: Molecular Cancer

Article Title: METTL3-mediated m 6 A modification of CACNA1E promotes osteosarcoma progression and chemoresistance by enhancing WNT7B-mediated Ca 2+ signaling

doi: 10.1186/s12943-025-02553-x

Figure Lengend Snippet: Targeted inhibition of CACNA1E improves MTX sensitivity and overcomes MTX resistance through WNT7B-mediated calcium signaling pathway. A - C P-gp and CACNA1E protein levels measured by Western blot in parental and MTX-resistant OS cells. D , E Cell viability of shNC-/shCACNA1E-transfected and MTX-treated OS cells. F , G Apoptosis of shNC-/shCACNA1E-transfected and MTX-treated OS cells analyzed by flow cytometry. H , I Colony-forming potential of shNC-/shCACNA1E-transfected and MTX-treated MTX-resistant OS cells. J - M IC50 value of MTX in shNC-/shCACNA1E-transfected MTX-resistant OS cells. N - Q Representative Chou-Talalay plots and CI-Fa plots showing the synergistic effects between CACNA1E knockdown and MTX analyzed using the CompuSyn software. R , S Apoptosis of shNC-/shCACNA1E-transfected and MTX-treated MTX-resistant OS cells measured by TUNEL staining. Scale bar, 100 μm. T , U Intracellular Ca 2+ level in shNC-/shCACNA1E-transfected and MTX-treated MTX-resistant OS cells detected by Fluo-8 AM calcium assay. Scale bar, 100 μm. V , W WNT7B protein level measured by Western blot in the above cells. These data are presented as the means ± SD of three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001 derived from Student’s t-test or one- or two-way analysis of variance

Article Snippet: MTX-resistant OS cell lines (MNNG MR and SJSA-1 MR ) were generated by gradually increasing the concentration of MTX (HY-14519, MCE, NJ, USA), as previously reported [ ].

Techniques: Inhibition, Western Blot, Transfection, Flow Cytometry, Knockdown, Software, TUNEL Assay, Staining, Calcium Assay, Derivative Assay

Journal: Cell Death & Disease

Article Title: Nanoparticle enhanced MRI can monitor macrophage response to CD47 mAb immunotherapy in osteosarcoma

doi: 10.1038/s41419-018-1285-3

Figure Lengend Snippet: a CD47 gene expression in a normal bone cell line, a normal patient bone, and osteoma patient specimen, three different osteosarcoma cell lines and eight osteosarcoma patient specimens. CD47 expression was measured with quantitative real-time PCR (qPCR), using glyceraldehyde 3-phosphate dehydrogenase (GAPDH) as an endogenous control, and compared between osteosarcomas and osteomas and normal bone using exact two-sided Wilcoxon rank-sum tests, p < 0.001. b Representative confocal CD47, CD68, and CD163 stains of three human osteosarcoma samples (scale bar 25 μm), a human osteoma sample (scale bar 25 μm), and a human bone sample (scale bar 50 μm) as negative control. c Corresponding quantitative area of CD47, CD68, and CD163 stains of one bone, one osteoma, and five osteosarcoma samples was measured over four high-power fields with Image J. Differences in CD47-, CD68-, and CD163-stained area of osteosarcoma, osteoma, and bone specimens were tested with exact two-sided Wilcoxon rank-sum tests, p < 0.001

Article Snippet: In vitro studies were performed in three human osteosarcoma cell lines (Saos-2, U-2 OS, MNNG/HOS, ATCC, Manassas, VA, USA), and one murine osteosarcoma cell line (K7M2, ATCC, Manassas, VA, USA).

Techniques: Gene Expression, Expressing, Real-time Polymerase Chain Reaction, Control, Negative Control, Staining

Journal: Cell Death & Disease

Article Title: Nanoparticle enhanced MRI can monitor macrophage response to CD47 mAb immunotherapy in osteosarcoma

doi: 10.1038/s41419-018-1285-3

Figure Lengend Snippet: a Schematic representation of experimental design. Tumors were initiated in NSG mice ( n = 6/group) by subcutaneous injection of human MNNG/HOS osteosarcoma cells positive for tdTomato-Luciferase expression. Treatment with control and anti-CD47 mAbs (10 mg/kg, 3× for 5 days) was initiated once the tumors were detected with bioluminescent imaging. MRI was performed on day 5, day 6, and day 10 of therapy. b Representative T2-weighted MR images of MNNG/HOS subcutaneous tumors in mice treated with control or CD47 mAbs. The tumors are hyperintense (bright) on pre-contrast MR images (white arrows) and show hypointense (dark) enhancement after ferumoxytol administration (red arrows). c Tumor MRI enhancement, quantified as T2 relaxation times, of MNNG/HOS tumors treated with control IgG or CD47 mAbs. CD47 mAb-treated tumors demonstrated significantly shortened T2 relaxation times compared to control antibody-treated tumors on ferumoxytol-enhanced MRI images. d Tumor volumes, as measured on T2-weighted MR scans, were significantly smaller on day 10 after anti-CD47 mAb treatment compared to controls. All results are represented as mean ± SD from six tumors per experimental group, p value as indicated, exact two-sided Wilcoxon rank-sum tests

Article Snippet: In vitro studies were performed in three human osteosarcoma cell lines (Saos-2, U-2 OS, MNNG/HOS, ATCC, Manassas, VA, USA), and one murine osteosarcoma cell line (K7M2, ATCC, Manassas, VA, USA).

Techniques: Injection, Luciferase, Expressing, Control, Imaging

Journal: Cell Death & Disease

Article Title: Nanoparticle enhanced MRI can monitor macrophage response to CD47 mAb immunotherapy in osteosarcoma

doi: 10.1038/s41419-018-1285-3

Figure Lengend Snippet: a Representative coronal T2-weighted MR images of intratibial K7M2 osteosarcomas at different time points after therapy with control or CD47 mAbs. White arrow points tumors on pre-contrast images, and red arrow points on post-contrast images. b T2 relaxation times of K7M2 osteosarcomas on unenhanced and ferumoxytol-enhanced MRI images at day 6 after control or CD47 mAb therapy. Only ferumoxytol-enhanced MR images show a significantly different T2 relaxation time between CD47 mAb-treated tumors and controls. c Prussian blue iron stains and F4/80, CD80, inducible nitric oxide synthase (iNOS) TAM stains, and active caspase-3 stain of K7M2 intratibial tumors treated with control and CD47 mAb. d Corresponding quantitative area of Prussian blue iron staining and F4/80, CD80, iNOS TAM staining, and caspase-3 staining in control and CD47 mAb-treated sets. e Bioluminescent in vivo images of mice with intratibial K7M2 osteosarcomas before and after therapy with IgG or CD47 mAb. f Total quantified flux in control and treated mice with K7M2 osteosarcomas. Results are represented as mean ± SD from six tumors per experimental group, p value as indicated, exact two-sided Wilcoxon rank-sum tests

Article Snippet: In vitro studies were performed in three human osteosarcoma cell lines (Saos-2, U-2 OS, MNNG/HOS, ATCC, Manassas, VA, USA), and one murine osteosarcoma cell line (K7M2, ATCC, Manassas, VA, USA).

Techniques: Control, Staining, In Vivo

Journal: Cell Death & Disease

Article Title: Nanoparticle enhanced MRI can monitor macrophage response to CD47 mAb immunotherapy in osteosarcoma

doi: 10.1038/s41419-018-1285-3

Figure Lengend Snippet: The signal-regulatory protein alpha (SIRPα) surface marker on osteosarcoma cells inhibits macrophage phagocytosis. Blocking the CD47–SIRPα interaction with CD47 monoclonal antibody (mAb) induces phagocytosis of both tumor and ferumoxytol nanoparticles. Increased uptake of ferumoxytol nanoparticles generates T2 contrast on MR scans that can serve as an imaging biomarker for monitoring responses to CD47 immunotherapy

Article Snippet: In vitro studies were performed in three human osteosarcoma cell lines (Saos-2, U-2 OS, MNNG/HOS, ATCC, Manassas, VA, USA), and one murine osteosarcoma cell line (K7M2, ATCC, Manassas, VA, USA).

Techniques: Marker, Blocking Assay, Imaging, Biomarker Discovery