Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: OS-derived exosomes induce lung fibroblasts activation and promote OS lung migration. A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis. Scale bar: 100 nm. C . Western blot analysis was performed to examine the expression of marker proteins in exosomes derived from MG63 and HOS cells. D . Confocal imaging indicates that Dil-labeled exosomes (red) were delivered to DAPI-labeled HFL-1 cells (blue). The green arrow indicates the delivered exosomes and representative images. Scale bar: 25 μm. E. qRT-PCR detection of the expression of α-SMA in the exosomes-stimulated HFL-1 cells. F. Schematic diagram of cell co-culture in-vitro model. G and H. Representative images and quantitative analysis of transwell assay of OS cells migrated to HFL-1 cells when stimulated with OS-derived exosomes, respectively. I. Representative images of lung metastasis of mice stimulated with MG63-derived exosomes. J. qRT-PCR detection of the relative expression of IL-1β, IL-6, and IL-8 in HFL-1 cells stimulated with the two types of exosomes. (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Derivative Assay, Activation Assay, Migration, Electron Microscopy, Western Blot, Expressing, Marker, Imaging, Labeling, Quantitative RT-PCR, Co-Culture Assay, In Vitro, Transwell Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: OS-derived exosomal linc00881 promotes OS lung migration and induces lung fibroblasts activation. A . RNA sequencing in a heatmap. B , C . Scatter plots of lncRNA expression in different groups. D . Relative expression of linc00881 in HFL-1 cells simulated by two OS exosomes. E . qRT-PCR detection of the expression linc00881 in exosomes with inhibited or overexpressed linc00881 in MG63 or HOS cells. F . Schematic diagram of the cell co-culture in vitro model. G – J . Representative images of MG63 ( G ) and HOS ( I ) cells migrated to HFL-1 cells with overexpressed or inhibited linc00881 in MG63 or HOS exosomes by transwell assay and the corresponding quantitative analysis ( H , J ). K – N . Representative images of MG63 ( K ) and HOS ( M ) cells migrated to HFL-1 cells with or without an inhibited expression of linc00881 in HFL-1 cells by transwell assay and the corresponding quantitative analysis ( L , N ). O , P . qRT-PCR detection of the relative expression of IL-1β, IL-6, IL-8, and α-SMA in HFL-1 cells with inhibited expression of linc00881 in MG63 or HOS exosomes. (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Derivative Assay, Migration, Activation Assay, RNA Sequencing, Expressing, Quantitative RT-PCR, Co-Culture Assay, In Vitro, Transwell Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: Linc00881 sponge miR-29c-3p in lung fibroblasts. A . Distribution of linc00881 in HFL-1 cells assessed by the FISH assay. B . Duplex structure of miR-29c-3p and linc00881. C . Relative expression of miR-29c-3p in HFL-1 cells simulated by two OS exosomes. D . qRT-PCR detection of relative expression of linc00881 using the control or miR-29c-3p probe to perform the pull-down assay. E . qRT-PCR detection of the expression of linc00881 in HFL-1 cells transfected with linc00881 siRNA or vector. F . qRT-PCR detection of the expression of miR-29c-3p in HFL-1 cells transfected as described in E. G – K . Transwell assay of MG63 or HOS cells migrated to the HFL-1 cells transfected with: a. control mimic + control vector; b. miR-29c-3p mimic + control vector; c. control mimic + linc00881 vector; d. miR-29c-3p mimic + linc00881 vector, representative images and quantitative analysis of MG63 and HOS cells are shown in H&I and J&K, respectively. (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Expressing, Quantitative RT-PCR, Control, Pull Down Assay, Transfection, Plasmid Preparation, Transwell Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: miR-29c-3p directly regulates the expression of MMP2 in lung fibroblasts cells. A . Duplex structure of MMP2 3′-UTR (Untranslated Region) and miR-29c-3p. B . The relative expression of MMP2 was detected at the mRNA level by qRT-PCR using a control or miR-125a-5p probe to perform the pull-down assay. C . The miR-29c-3p expression was detected via qRT-PCR after the control/miR-29c-3p mimic or control/miR-29c-3p inhibitor was transfected into HFL-1 cells. D – F . Representative images ( D ) and quantitative analysis ( E , F ) of Western blotting analysis and qRT-PCR of MMP2 expression in HFL-1 cells after transfected as described in C . G , H . Representative images ( G ) and quantitative analysis ( H ) of Western blotting analysis of MMP2 expression in HFL-1 cells transfected as follows: a. control mimic and control vector; b. control mimic and the miR-29c-3p-overexpressed plasmid; c. miR-29c-3p mimic and control vector; d. miR-29c-3p mimic and MMP2 vector. I – L . Representative images ( I MG63, K HOS) and quantitative analysis ( J , L ) of transwell assay of MG63 and HOS cells migrated to HFL-1 cells after transfected as described in G . (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Expressing, Quantitative RT-PCR, Control, Pull Down Assay, Transfection, Western Blot, Plasmid Preparation, Transwell Assay

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: Exosomal Linc00881 regulates the expression of MMP2 in lung fibroblasts by sponging miR-29c-3p. A , B . Representative images ( A ) of Western blotting analysis of MMP2 expression in HFL-1 cells treated with blank, MG63, or HOS exosomes and the corresponding quantitative analysis ( B ). C . Relative miR-29c-3p concentration was detected in HFL-1 cells simulated by the two types of OS-derived exosomes. D - F . Representative images ( D ) of Western blotting analysis of MMP2 expression in HFL-1 cells transfected with linc00881 siRNA or vector and the corresponding quantitative analysis ( E , F ). G . qRT-PCR detection of the expression of linc00881 in HFL-1 cells transfected with linc00881 siRNA or vector. H . qRT-PCR detection of the expression of miR-29c-3p in HFL-1 cells transfected as described in B . I – K . Representative images ( I ) of Western blotting analysis of MMP2 expression in HFL-1 cells transfected with miR-29c-3p mimic and stimulated with MG63 or HOS cells-derived exosomes, and the corresponding quantitative analysis ( J , K ). L – O . qRT-PCR detection of the relative expression of linc00881 or miR-29c-3p in HFL-1 cells treated in I . (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Expressing, Western Blot, Concentration Assay, Derivative Assay, Transfection, Plasmid Preparation, Quantitative RT-PCR

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: Linc00881 activates lung fibroblasts through the NF-κB axis. A , B . Western blotting analysis of the proteins of NF-κB axis in HFL-1 cells treated with exosomes from MG63 and HOS cells. C , D . Western blotting analysis of the expression of MMP2 and N-cadherin in HFL-1 cells treated with MG63 or HOS cell-derived exosomes. E , F . Western blotting analysis of the proteins of NF-κB axis in HFL-1 cells treated with: a. control vector, b. linc00881 vector, c. control siRNA, d. MMP2 siRNA, e. control MMP2, and f. MMP2 vector. G , H . Western blotting analysis of the expression of MMP2 and N-cadherin in HFL-1 cells in different treatments as described in E . (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Western Blot, Expressing, Derivative Assay, Control, Plasmid Preparation

Journal: Cancer Cell International

Article Title: Tumor-derived exosomal linc00881 induces lung fibroblast activation and promotes osteosarcoma lung migration

doi: 10.1186/s12935-023-03121-3

Figure Lengend Snippet: Activated fibroblasts by exosomal linc00881 accelerate OS progression. A . IL-6 secretion from HFL-1 treated by exosomes from two OS cells was detected via ELISA assay. B . IL-6 secretion from HFL-1 treated by exosomes from two OS cells or exosomes from linc00881 interrupted two OS cells and was detected via ELISA assay. C . Spheroid formation ability of MG63 treated with indicated CM. Representative Images and quantitative analysis were shown.100 × . D – E . Spheroid formation ability of MG63 treated with CM containing: a. IL-6 neutralizing antibody, b. IgG control antibody, c. linc00881 siRNA exo, d. IL-6 neutralizing antibody plus linc00881 siRNA exo. Representative Images and quantitative analysis were shown.100 × . F . Spheroid formation ability of HOS treated with indicated CM. Representative Images and quantitative analysis were shown.100 × . G . Spheroid formation ability of MG63 treated with CM as indicated in D . Representative Images and quantitative analysis were shown.100 × . H . Migration assay of MG63 treated with indicated CM. Representative Images and quantitative analysis were shown.100 × . I . Migration assay of MG63 treated with indicated CM indicate in D . Representative Images and quantitative analysis were shown.100 × . J , K . Migration assay of HOS treated with indicated CM. Representative Images and quantitative analysis were shown.100 × . L . Migration assay of HOS treated with CM indicated in D . Representative Images and quantitative analysis were shown.100 × . M , N . Western blotting analysis of the expression of MMP2 in MG63 and HOS cells treated with indicated CM. (* p < 0.05; ** p < 0.01; *** p < 0.0001)

Article Snippet: A , B . Exosomes from MG63 and HOS cells were measured using electron microscopy and Nanosight particle tracking analysis.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Migration, Western Blot, Expressing

Journal: International Journal of Biological Sciences

Article Title: Vitamin K 2 Ameliorates Damage of Blood Vessels by Glucocorticoid: a Potential Mechanism for Its Protective Effects in Glucocorticoid-induced Osteonecrosis of the Femoral Head in a Rat Model

doi: 10.7150/ijbs.15248

Figure Lengend Snippet: VEGF expression analysis in EAhy926 and MG63. A. Western blotting analysis of total cell lysates in EAhy926, β-actin was used as the loading control. B. VEGF mRNA expression in EAhy926 treated with or without DEX and VK 2 , each condition was performed in triplicate (* indicated P < 0.05; ** indicated P < 0.01). C. Western blotting analysis of total cell lysates in MG63. β-actin was used as the loading control. D. VEGF mRNA expression in MG63 cells treated with or without DEX and VK 2 , each condition was performed in triplicate (* indicated P < 0.05; ** indicated P < 0.01).

Article Snippet: The human endothelial cell line EAhy926 and the human osteoblast-like cell line MG63 were obtained from KeyGENE BIOTECH (Nanjing, China).

Techniques: Expressing, Western Blot, Control

Journal: International Journal of Biological Sciences

Article Title: Vitamin K 2 Ameliorates Damage of Blood Vessels by Glucocorticoid: a Potential Mechanism for Its Protective Effects in Glucocorticoid-induced Osteonecrosis of the Femoral Head in a Rat Model

doi: 10.7150/ijbs.15248

Figure Lengend Snippet: Effect of DEX and VK 2 on angiogenic factors in EAhy926 and MG63. (A-E) Quantitative RT-PCR analysis of KDR, Flt, vWF, CD31 and PDGF-B in EAhy926. (F,G) TGF-β and BMP-2 mRNA expression in MG63. (Each condition was performed in triplicate, * indicated P < 0.05, ** indicated P < 0.01).

Article Snippet: The human endothelial cell line EAhy926 and the human osteoblast-like cell line MG63 were obtained from KeyGENE BIOTECH (Nanjing, China).

Techniques: Quantitative RT-PCR, Expressing

Journal: Scientific Reports

Article Title: Sulfasalazine induces ferroptosis in osteosarcomas by regulating Nrf2/SLC7A11/GPX4 signaling axis

doi: 10.1038/s41598-025-13324-5

Figure Lengend Snippet: SAS inhibits OS cell viability. ( A ) Chemical structure of SAS. ( B ) Viability of MG63 and U2OS cells treated with SAS for 24, 48, and 72 h, as determined by CCK-8 assay. ( C , D ) Statistical analysis comparing the effects of varying SAS treatment durations at a fixed concentration ( C ) and varying SAS concentrations at a fixed duration ( D ) on the viability of MG63 and U2OS cells. (Data were presented as mean ± SD, n = 3; * P < 0.05. ns, no statistical significance.)

Article Snippet: MG63 cells, obtained from Servicebio (Wuhan, China), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with non-essential amino acids (NEAA).

Techniques: CCK-8 Assay, Concentration Assay

Journal: Scientific Reports

Article Title: Sulfasalazine induces ferroptosis in osteosarcomas by regulating Nrf2/SLC7A11/GPX4 signaling axis

doi: 10.1038/s41598-025-13324-5

Figure Lengend Snippet: Effect of SAS on OS Cell Behavior. ( A ) EDU staining assay showing the effect of 24-h SAS treatment on MG63 and U2OS cell proliferation. ( B ) Cell cycle distribution in MG63 and U2OS cells following 24-h SAS treatment, as determined by flow cytometry. Statistical analysis was performed to quantify the proportion of cells in the G1 phase across all experimental groups. ( C ) Representative images of MG63 and U2OS cells treated with various concentrations of SAS for 24 h, illustrating the effect on cell migration. ( D ) Quantification of cell migration in MG63 and U2OS cells following 24-h treatment with various concentrations of SAS. ( E ) Apoptosis rate in MG63 and U2OS cells following 24-h SAS treatment, as determined by flow cytometry. (Data were presented as mean ± SD, n = 3. * P < 0.05 versus control group.)

Article Snippet: MG63 cells, obtained from Servicebio (Wuhan, China), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with non-essential amino acids (NEAA).

Techniques: Staining, Flow Cytometry, Migration, Control

Journal: Scientific Reports

Article Title: Sulfasalazine induces ferroptosis in osteosarcomas by regulating Nrf2/SLC7A11/GPX4 signaling axis

doi: 10.1038/s41598-025-13324-5

Figure Lengend Snippet: SAS induced cell death via triggering ferroptosis. ( A ) Effect of 24-h treatment with ferroptosis inhibitors (DFO, Fer-1, Lip-1) on the viability of MG63 and U2OS cells, as determined by CCK-8 assay. ( B ) Western blot analysis of FTH1 protein expression in MG63 and U2OS cells following 24-h SAS treatment. ( C ) Intracellular Fe2 + levels in MG63 and U2OS cells following 24-h SAS treatment. ( D ) Mitochondrial membrane potential (MMP) in MG63 and U2OS cells following 24-h SAS treatment, as determined by flow cytometry. (Data were presented as mean ± SD, n = 3; * P < 0.05.)

Article Snippet: MG63 cells, obtained from Servicebio (Wuhan, China), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with non-essential amino acids (NEAA).

Techniques: CCK-8 Assay, Western Blot, Expressing, Membrane, Flow Cytometry

Journal: Scientific Reports

Article Title: Sulfasalazine induces ferroptosis in osteosarcomas by regulating Nrf2/SLC7A11/GPX4 signaling axis

doi: 10.1038/s41598-025-13324-5

Figure Lengend Snippet: SAS induces ferroptosis in OS Cells. ( A ) Glutathione (GSH) levels in MG63 and U2OS cells following 24-h SAS treatment. ( B ) Superoxide dismutase (SOD) levels in MG63 and U2OS cells following 24-h SAS treatment. ( C ) Malondialdehyde (MDA) levels in MG63 and U2OS cells following 24-h SAS treatment. ( D ) Reactive oxygen species (ROS) levels in MG63 and U2OS cells following 24-h SAS treatment, as determined by flow cytometry. (Data were presented as mean ± SD, n = 3. * P < 0.05 versus control group.)

Article Snippet: MG63 cells, obtained from Servicebio (Wuhan, China), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with non-essential amino acids (NEAA).

Techniques: Flow Cytometry, Control

Journal: Scientific Reports

Article Title: Sulfasalazine induces ferroptosis in osteosarcomas by regulating Nrf2/SLC7A11/GPX4 signaling axis

doi: 10.1038/s41598-025-13324-5

Figure Lengend Snippet: Molecular Mechanisms of SAS-Induced Effects on OS Cells . ( A ) Western blot analysis of NRF2, SLC7A11, and GPX4 protein expression in MG63 and U2OS cells following 24-h SAS treatment. ( B ) Western blot analysis of NRF2, SLC7A11, and GPX4 protein expression in MG63 and U2OS cells following treatment with RTA-480. ( C ) Quantitative PCR (qPCR) analysis of NRF2, SLC7A11, and GPX4 mRNA expression in MG63 and U2OS cells following 24-h SAS treatment. (Data were presented as mean ± SD, n = 3 ; * P < 0.05.)

Article Snippet: MG63 cells, obtained from Servicebio (Wuhan, China), were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with non-essential amino acids (NEAA).

Techniques: Western Blot, Expressing, Real-time Polymerase Chain Reaction