os cell lines saos 2  (ATCC)

Journal: Journal of Cellular and Molecular Medicine

Article Title: Gene Expression Alterations Associated With Resveratrol‐Induced Antiproliferative Effects and S‐Phase Cell Cycle Arrest in Osteosarcoma Cancer Cells

doi: 10.1111/jcmm.71111

Figure Lengend Snippet: Hyperspectral Imaging (HSI) of RSV and intracellular localisation in both OS cell lines. (A) Dark‐field optical imaging captures the signal of pure RSV, and (B) the associated spectral library shows the intensity of scattered light across wavelengths. (C) Dark‐field images and Spectral Angle Mapping (SAM) analysis in OS cell lines (SAOS‐2 and U2‐OS) show no RSV‐associated signals in control samples (completely black images), while treated cells display coloured pixels matching the RSV spectral signature, confirming intracellular localisation of the compound. Imaging was performed using a 60× oil immersion objective to resolve RSV distribution and spectral characteristics. (D) Quantification of RSV‐positive pixels reveals a statistically significant increase in both treated OS cell lines, compared to controls (*** p < 0.001), confirming cellular uptake of RSV.

Article Snippet: In vitro assays were conducted using OS cell lines SAOS‐2 and U2‐OS (ATCC, Cat. no. HTB‐85; Cat. no. HTB‐96) [ ], along with human bone marrow‐derived mesenchymal stem cells (hBMSCs) (Lonza Milan, Italy, PT‐2501). hBMSCs were characterised by flow cytometry analysis (FCA) for MSC surface markers, including positive markers (CD29, CD73 and CD90) and negative markers (CD14 and CD45) [ ].

Techniques: Imaging, Optical Imaging, Control

Journal: Journal of Cellular and Molecular Medicine

Article Title: Gene Expression Alterations Associated With Resveratrol‐Induced Antiproliferative Effects and S‐Phase Cell Cycle Arrest in Osteosarcoma Cancer Cells

doi: 10.1111/jcmm.71111

Figure Lengend Snippet: Effects of RSV on the proliferation, viability and cell cycle distribution of OS cell lines and hBMSCs. (A) The MTT assay was used to assess the effect of resveratrol (RSV, 1–1000 μM) on OS cell lines (SAOS‐2 and U2‐OS) and healthy hBMSCs over 24, 48 and 72 h. In SAOS‐2 cells, RSV reduced proliferation in a dose‐dependent manner, compared to control ( p < 0.001), with significant decreases at 100 and 1000 μM after 48 h, compared to other treatments (** p < 0.01). In U2‐OS cells, RSV significantly reduced viability at all concentrations, with more differences observed after 24 h at higher concentrations (*** p < 0.0001). hBMSCs showed no significant changes, except for a notable increase at 1000 μM (*** p < 0.0001). (B) The Live/Dead assay confirmed the cytotoxic effect of RSV (100 μM) on OS cells after 48 h, using green Cyto‐dye for live cells and red propidium iodide for dead cells. (C) Fluorescence image quantification showed a significant reduction in live cells (*** p < 0.0001) and an increase in dead cells (** p < 0.001; * p < 0.01) in RSV‐treated OS cell lines compared to the control. (D) The effect of RSV on the cell cycle was analysed by BrdU/PI staining and flow cytometry after 48 h of treatment with 100 μM. The cytogram displays that RSV treatment resulted in a significant accumulation of OS cells in the S phase and a decrease in the G0/G1 and G2/M phases, compared to untreated controls. (E) Statistical analysis confirmed a significant increase in the S phase (*** p < 0.0001) and a significant reduction in the G0/G1 phase (* p < 0.001; ***p < 0.0001) in both OS cell lines.

Article Snippet: In vitro assays were conducted using OS cell lines SAOS‐2 and U2‐OS (ATCC, Cat. no. HTB‐85; Cat. no. HTB‐96) [ ], along with human bone marrow‐derived mesenchymal stem cells (hBMSCs) (Lonza Milan, Italy, PT‐2501). hBMSCs were characterised by flow cytometry analysis (FCA) for MSC surface markers, including positive markers (CD29, CD73 and CD90) and negative markers (CD14 and CD45) [ ].

Techniques: MTT Assay, Control, Live Dead Assay, Fluorescence, Staining, Flow Cytometry

Journal: Journal of Cellular and Molecular Medicine

Article Title: Gene Expression Alterations Associated With Resveratrol‐Induced Antiproliferative Effects and S‐Phase Cell Cycle Arrest in Osteosarcoma Cancer Cells

doi: 10.1111/jcmm.71111

Figure Lengend Snippet: RSV induces apoptosis in OS cell lines. (A) Flow cytometry with Annexin V/PI staining was used to assess apoptosis in OS cells treated with 100 μM RSV for 48 h. The analysis identified early apoptotic (Annexin V+), late apoptotic (Annexin V+/PI+) and necrotic (PI+) cells, with Annexin V and PI intensities plotted on the X and Y axes, respectively. (B) Quantification showed that RSV significantly increased late apoptotic and necrotic cells in both OS cell lines (* p < 0.0001 for SAOS‐2 and ** p < 0.01 for U2‐OS), compared to control. A decrease in early apoptotic cells is observed in U2‐OS (** p < 0.01). (C) Gene expression profiling after RSV treatment revealed 18 differentially expressed apoptotic genes in SAOS‐2 (12 upregulated, 6 downregulated) and 21 in U2‐OS (10 upregulated, 11 downregulated), based on a Log 2 FC > 1 or < −1. (D) Gene Set Enrichment Analysis (GSEA) grouped these modulated genes into four categories: Positive regulators of apoptosis, negative regulators, caspases and death domain receptors, showing both up‐ and downregulated genes in each cell line. (E) A Venn diagram showed 11 apoptotic genes commonly modulated in both OS cell lines, with 7 unique to SAOS‐2 and 9 to U2‐OS, suggesting shared and cell‐specific mechanisms of RSV‐induced apoptosis. (F) Immunostaining for caspase‐3/7 revealed increased activation in both RSV‐treated OS cell lines after 48 h. (G) Fluorescence quantification using ImageJ confirmed a significant rise in activated caspase‐3/7 levels (** p < 0.001) in both RSV‐treated OS cells compared to controls.

Article Snippet: In vitro assays were conducted using OS cell lines SAOS‐2 and U2‐OS (ATCC, Cat. no. HTB‐85; Cat. no. HTB‐96) [ ], along with human bone marrow‐derived mesenchymal stem cells (hBMSCs) (Lonza Milan, Italy, PT‐2501). hBMSCs were characterised by flow cytometry analysis (FCA) for MSC surface markers, including positive markers (CD29, CD73 and CD90) and negative markers (CD14 and CD45) [ ].

Techniques: Flow Cytometry, Staining, Control, Gene Expression, Immunostaining, Activation Assay, Fluorescence

Journal: Journal of Cellular and Molecular Medicine

Article Title: Gene Expression Alterations Associated With Resveratrol‐Induced Antiproliferative Effects and S‐Phase Cell Cycle Arrest in Osteosarcoma Cancer Cells

doi: 10.1111/jcmm.71111

Figure Lengend Snippet: RSV inhibits cell migration and modulates ECM related gene expression in OS cells lines. (A) Bright‐field images from a wound healing assay show that control OS cells fully close the wound by 72 h (T3), whereas RSV‐treated cells (100 μM) exhibit no wound closure at any time point 0‐72 h (T0–T3), indicating that RSV strongly inhibits cell migration. (B) Quantitative analysis confirms significant wound closure in control cells over time compared to baseline T0 (0h) (° p < 0.0001), with additional increases at 48 h (T2) and 72 h (T3) compared to 24 h (T1) (* p < 0.001). Complete closure is observed at 72 h (T3) in control cells. (C) ECM‐related gene expression analysis using RT 2 Profiler PCR Array shows differential expression in RSV‐treated cells: 43 genes are modulated in SAOS‐2 (29 upregulated, 14 downregulated) and 26 in U2‐OS (11 upregulated, 15 downregulated). (D) Gene Set Enrichment Analysis (GSEA) categorizes these genes into five functional groups: Cell–cell adhesion, ECM‐cell adhesion, ECM constituents, ECM remodelling and basement membrane components. Up‐ and downregulated genes are identified for each group in both cell lines. (E) A Venn diagram reveals 20 ECM‐related genes commonly modulated in both SAOS‐2 and U2‐OS, indicating shared pathways influenced by RSV, particularly those involved in cell adhesion and ECM remodelling.

Article Snippet: In vitro assays were conducted using OS cell lines SAOS‐2 and U2‐OS (ATCC, Cat. no. HTB‐85; Cat. no. HTB‐96) [ ], along with human bone marrow‐derived mesenchymal stem cells (hBMSCs) (Lonza Milan, Italy, PT‐2501). hBMSCs were characterised by flow cytometry analysis (FCA) for MSC surface markers, including positive markers (CD29, CD73 and CD90) and negative markers (CD14 and CD45) [ ].

Techniques: Migration, Gene Expression, Wound Healing Assay, Control, Quantitative Proteomics, Functional Assay, Membrane

Journal: Journal of Cellular and Molecular Medicine

Article Title: Gene Expression Alterations Associated With Resveratrol‐Induced Antiproliferative Effects and S‐Phase Cell Cycle Arrest in Osteosarcoma Cancer Cells

doi: 10.1111/jcmm.71111

Figure Lengend Snippet: RSV modulates the Wnt/β‐Catenin signalling pathway and affects vimentin expression and β‐catenin localisation in OS cells lines. (A) Real‐time PCR analysis shows that RSV treatment (100 μM, 48 h) significantly downregulates key genes of the Wnt/β‐catenin pathway in SAOS‐2 and U2‐OS cells, including CTNNB1 , MMP7 , MMP9 and CD44 (* p < 0.001), all associated with ECM degradation, stemness and invasiveness. Conversely, CDH1 (epithelial marker) is upregulated in both lines (* p < 0.001 for SAOS‐2 and ** p < 0.01 for U2‐OS), suggesting a shift toward an epithelial phenotype. WNT1 and VIM are significantly downregulated (* p < 0.01 and * p < 0.05, respectively) in both cell lines, with c‐MYC reduced in SAOS‐2 (* p < 0.01). (B) Immunocytochemistry reveals a notable decrease in Vimentin protein levels in RSV‐treated cells. Vimentin, a mesenchymal marker, appears less expressed, with treated cells showing morphological changes including elongated filaments and enlarged cell body and nucleus. (C) β‐catenin immunostaining indicates that RSV prevents its nuclear translocation, with the protein mainly localized at cell junctions and in the cytoplasm in treated cells, while in controls β‐catenin is predominantly nuclear, confirming RSV‐mediated inhibition of Wnt/β‐catenin signalling.

Article Snippet: In vitro assays were conducted using OS cell lines SAOS‐2 and U2‐OS (ATCC, Cat. no. HTB‐85; Cat. no. HTB‐96) [ ], along with human bone marrow‐derived mesenchymal stem cells (hBMSCs) (Lonza Milan, Italy, PT‐2501). hBMSCs were characterised by flow cytometry analysis (FCA) for MSC surface markers, including positive markers (CD29, CD73 and CD90) and negative markers (CD14 and CD45) [ ].

Techniques: Expressing, Real-time Polymerase Chain Reaction, Marker, Immunocytochemistry, Immunostaining, Translocation Assay, Inhibition

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