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: 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: The American Journal of Pathology

Article Title: Chemokine (C-C Motif) Ligand 2/CCR2/Extracellular Signal-Regulated Kinase Signal Induced through Cancer Cell–Macrophage Interaction Contributes to Hepatocellular Carcinoma Progression

doi: 10.1016/j.ajpath.2024.12.007

Figure Lengend Snippet: Co-culturing with macrophages facilitates the malignant phenotypes of hepatocellular carcinoma (HCC) cells. A: Experimental scheme of establishment of peripheral blood monocyte-derived macrophages. Peripheral blood mononuclear cells (PBMCs) are collected from healthy donors. Peripheral blood CD14-positive monocytes (PBMos) are purified from PBMCs using autoMACS pro Separator and incubated with recombinant human macrophage colony-stimulating factor (rhM-CSF) and granulocyte-macrophage colony-stimulating factor (rhGM-CSF) for 6 days, introducing PBMo-derived macrophages (M0). B: Experimental scheme of transwell proliferation assay of HCC cells (Hep G2, HuH-7, and HLE) and M0. HCC cells (4.0 × 10 4 cells per well) are seeded in lower plates. For the co-culture system, M0 (3.0 × 10 4 cells per well) are seeded in upper chambers (0.4-μm pore). After 48 hours, dimethylthiazol-carboxymethoxyphenyl-sulfophenyl-tetrazolium (MTS) assays are performed to compare the cell proliferation between monocultured (Mono) and co-cultured (Co) HCC cells. C: Experimental scheme of transwell migration assay of HCC cells (Hep G2, HuH-7, and HLE) and M0. HCC cells (1.0 × 10 5 cells per well) are seeded in upper chambers (8.0-μm pore). For the co-culture system, M0 (8.0 × 10 4 cells per well) are seeded in lower plates. After 48 hours, HCC cells migrating to the lower surface are counted to compare the cell migration between monocultured and co-cultured HCC cells. D: Representative images of transwell migration assays shown in C . E: Expression levels of Akt, phosphorylated Akt (p-Akt), extracellular signal-regulated kinase (Erk), phosphorylated Erk (p-Erk), NF-κB, and phosphorylated NF-κB (p-NF-κB) in monocultured and co-cultured HCC cells based on Western blot analysis. β-Actin is used as a loading control. F: Quantification of the Western blot analysis bands illustrated in E . The expression levels of p-Akt (Ser473 and Thr308), p-Erk (Thr202/Tyr204), and p-NF-κB in monocultured and co-cultured HCC cells with M0 are normalized to the respective total protein levels. B , C , and E: Experiments are performed in triplicate and repeated at least thrice. B , C , and F: Two-sided t -tests were performed for the statistical analysis of parametric data. Data are expressed as the means ± SEM ( B , C , and F ). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P < 0.001. Scale bars = 100 μm ( D ). DMEM, Dulbecco’s modified Eagle’s medium; NS, not significant.

Article Snippet: Three human HCC cell lines, Hep G2 (hepatoblastoma cell line, classically used as HCC cell line), HuH-7 (well-differentiated HCC cell line), and HLE (undifferentiated HCC cell line), were obtained from RIKEN BioResource Center (Tsukuba, Japan) and cultured in low-glucose Dulbecco's modified Eagle's medium (DMEM; Fujifilm, Osaka, Japan) supplemented with 10% fetal bovine serum (Sigma-Aldrich) and 1% antibiotic-antimycotic (Invitrogen, Carlsbad, CA).

Techniques: Derivative Assay, Purification, Incubation, Recombinant, Proliferation Assay, Co-Culture Assay, Cell Culture, Transwell Migration Assay, Migration, Expressing, Western Blot, Control, Modification