Journal: Bone Research

Article Title: ADAR2 induces the differentiation of osteosarcoma cells by editing activity on IGFBP7: new implications for therapy

doi: 10.1038/s41413-026-00516-6

Figure Lengend Snippet: Effects of ADAR2 overexpression in osteosarcoma cell lines. a Real-Time RT-PCR and b Western Blot analysis of ADAR2 expression in MSC, osteoblasts (OB) and osteosarcoma cell lines Saos-2 and 143B. In b upper panels : representative blots; lower panel : densitometric analysis. c FACS analysis of the proliferative rate evaluated by CMAC staining and d cell cycle analysis of Saos-2 ( left panel ) and 143B ( right panel ) cells transfected with ADAR2-pEGFP-C3 (pADAR2), ADAR2 E/A-pEGFP-C3 (pADAR2 E/A) or Empty-pEGFP-C3 (pEmpty) vectors. ADAR2 E/A vector was generated by a single mutation in the catalytic domain of ADAR2. e Migration ability of transfected Saos-2 ( left panel) and 143B ( right panel) cells. f Transwell invasion assay of transfected Saos-2 ( left panel ) and 143B cells ( right panel ). g Representative blot and h densitometric analysis of Runx2 and Osx in transfected Saos-2 ( left panel) and 143B cells ( right panel ). Results are expressed as mean ± sd and are reported as individual data points of independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; vs pEmpty transfected cells. # P < 0.05; ## P < 0.01 vs pADAR2 transfected cells

Article Snippet: Commercially available human OS cell lines Saos-2 (HTB-85) and 143B (CRL-8303) were purchased from American Type Culture Collection (ATCC, Washington, NW, USA).

Techniques: Over Expression, Quantitative RT-PCR, Western Blot, Expressing, Staining, Cell Cycle Assay, Transfection, Plasmid Preparation, Generated, Mutagenesis, Migration, Transwell Invasion Assay

Journal: Bone Research

Article Title: ADAR2 induces the differentiation of osteosarcoma cells by editing activity on IGFBP7: new implications for therapy

doi: 10.1038/s41413-026-00516-6

Figure Lengend Snippet: Terminal osteogenic differentiation and increased drugs susceptibility in pADAR2-transfected Saos-2 cells. a – c Mineralization assay of Saos-2 cells transfected with pADAR2, pADAR2 E/A or pEmpty vectors. a Upper panels : Alizarin Red staining; lower panels : Von Kossa staining. b Absorbance analysis of Alizarin Red staining. c Densitometric analysis of Von Kossa-stained area. d – h Real-Time RT-PCR expression analysis of d COL1A2 , e DMP1 , f MEPE , g PRKCA and h NANOG . In ( b – h ) results are expressed as mean ± sd and are reported as individual data points of independent experiments. i Cell viability analysis of transfected Saos-2 cells treated for 6 days with increasing concentrations of MTX (0, 1, 5, 10, 50 and 100 nmol/L, left panel ) and of MS275 (0, 0.5, 1, 2.5, 5 and 10 μmol/L, right panel ) for 2 days. The concentration of drugs able to reduce by 50% (GI 50 ) cell viability is reported in the upper part of each graph. Results are expressed as mean ± sd of at least three independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.000 1 vs pEmpty transfected cells. # P < 0.05; ## P < 0.01; ### P < 0.001; #### P < 0.000 1 vs pADAR2 transfected cells. j FACS analysis of apoptosis of transfected Saos-2 cells treated with the GI 50 calculated for pEmpty transfected Saos-2, or with Vehicle. Results are expressed as mean ± sd and are reported as individual data points of independent experiments. ** P < 0.01 vs Vehicle treated cells

Article Snippet: Commercially available human OS cell lines Saos-2 (HTB-85) and 143B (CRL-8303) were purchased from American Type Culture Collection (ATCC, Washington, NW, USA).

Techniques: Transfection, Mineralization Assay, Staining, Quantitative RT-PCR, Expressing, Concentration Assay

Journal: Bone Research

Article Title: ADAR2 induces the differentiation of osteosarcoma cells by editing activity on IGFBP7: new implications for therapy

doi: 10.1038/s41413-026-00516-6

Figure Lengend Snippet: In vivo experiments. Seven-weeks-old NSG male mice were intratibially injected with Saos-2 cells transfected with pADAR2, pADAR2 E/A or pEmpty vectors; after 12 weeks animals were sacrificed. a Representative X-Ray pictures of primary bone tumors. b Quantification of the tumor volume. c Number of animals with metastases in liver, lungs and kidneys at sacrifice. d Hematoxylin/Eosin staining of liver, lungs and kidney metastases. Nodules were indicated by black arrowheads. Number of metastases in e liver, f lungs and g kidneys. h Representative pictures of immunohistochemistry and i quantification of Ki67 in liver, lungs and kidneys metastases. Results are expressed as mean ± sd. * P < 0.05; ** P < 0.01 vs pEmpty cells injected mice. ## P < 0.01 vs pADAR2 cells injected animals

Article Snippet: Commercially available human OS cell lines Saos-2 (HTB-85) and 143B (CRL-8303) were purchased from American Type Culture Collection (ATCC, Washington, NW, USA).

Techniques: In Vivo, Injection, Transfection, Staining, Immunohistochemistry

Journal: Bone Research

Article Title: ADAR2 induces the differentiation of osteosarcoma cells by editing activity on IGFBP7: new implications for therapy

doi: 10.1038/s41413-026-00516-6

Figure Lengend Snippet: RNA-seq analysis. a Gene expression based heatmap showing the unique clusterization of ADAR2 transfected Saos-2 cells. Real-Time RT-PCR expression analysis of b COL4A1 , c SERPINH1 , d SWAP-70 and e TENM1 for transcriptional validation. f – h Editing analysis. Upper panels : Sequence chromatograms of the transcripts and editing levels of COPA , IGFBP7 and COG3 . Arrows indicate editing positions. Lower panels : percentage of editing in f COPA , g IGFBP7 and h COG3 transcripts. Results are expressed as mean ± sd and are reported as individual data points of independent experiments. * P < 0.05; ** P < 0.01; **** P < 0.000 1 vs pEmpty transfected cells. # P < 0.05; ## P < 0.01; ### P < 0.001 vs pADAR2 transfected cells

Article Snippet: Commercially available human OS cell lines Saos-2 (HTB-85) and 143B (CRL-8303) were purchased from American Type Culture Collection (ATCC, Washington, NW, USA).

Techniques: RNA Sequencing, Gene Expression, Transfection, Quantitative RT-PCR, Expressing, Biomarker Discovery, Sequencing

Journal: Bone Research

Article Title: ADAR2 induces the differentiation of osteosarcoma cells by editing activity on IGFBP7: new implications for therapy

doi: 10.1038/s41413-026-00516-6

Figure Lengend Snippet: IGF1R pathway analysis. a – f Investigation of IGF1R pathway in Saos-2 cells transfected with pADAR2, pADAR2 E/A or pEmpty vectors. a Representative plots and b – f densitometric analysis of b p-Igf1r, c p-Irs, d p-Akt (T308), e p-Akt (S473) and f p-p70. Results are expressed as mean ± sd and are reported as individual data points of independent experiments. ** P < 0.01; *** P < 0.00 1 vs pEmpty transfected cells. # P < 0.05; ## P < 0.01 vs pADAR2 transfected cells. g – o Effects of the treatment with WT- or K95R-IGFBP7 on Saos-2 cell line. g – l Analysis of IGF1R pathway in Saos-2 cells treated with 2 μg/ml of WT- or K95R-IGFBP7 compared to vehicle (Veh) treated cells. g Representative plots and h – l densitometric analysis of h p-Igf1r, i p-Irs, j p-Akt (T308), k p-Akt (S473) and l p-p70. m , n Western blot analysis of Runx2 in Saos-2 cells treated with WT- or K95R-IGFBP7 compared to vehicle (Veh) treated cells. m Representative blot and n densitometric analysis. o Proliferation rate of Saos-2 treated with WT- or K95R-IGFBP7 compared to vehicle (Veh) treated cells. Results are expressed as mean ± sd and are reported as individual data points of independent experiments. * P < 0.05; ** P < 0.01; *** P < 0.001 vs Vehicle treated Saos-2 cells. # P < 0.05; ### P < 0.001 vs WT-IGFBP7 treated cells

Article Snippet: Commercially available human OS cell lines Saos-2 (HTB-85) and 143B (CRL-8303) were purchased from American Type Culture Collection (ATCC, Washington, NW, USA).

Techniques: Transfection, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Creatine Kinase Blockade Disrupts Energy Metabolism and Redox Homeostasis to Suppress Osteosarcoma Progression

doi: 10.3390/ijms262311555

Figure Lengend Snippet: Effects of CK inhibition on stemness. ( A ) Sphere-forming capacity in SaOS2 and U2OS cells following DNFB treatment. Images were taken with a phase-contrast microscope. Scale bar, 100 μm. ( B ) Expression of stemness markers Oct3 and Nestin assessed by RT-PCR. * p < 0.05 vs. C or DNFB(−). Error bars: standard deviation of three independent trials. Statistical differences were calculated using ordinary ANOVA with Bonferroni correction. ANOVA, analysis of variance; DNFB, dinitrofluorobenzene; CK, creatine kinase; OCT3, POU Class 5 homeobox 1; ACTB, β-actin.

Article Snippet: Human OS cell lines SaOS2, U2OS, HOS, MG63, and human bone marrow-derived mesenchymal stem cell line (hMSC, PCS-500-01) were purchased from the American Type Culture Collection (ATCC; Rockville, MD, USA).

Techniques: Inhibition, Microscopy, Expressing, Reverse Transcription Polymerase Chain Reaction, Standard Deviation