Journal: Scientific Reports

Article Title: microRNA-29b mediates fibrotic induction of human xylosyltransferase-I in human dermal fibroblasts via the Sp1 pathway

doi: 10.1038/s41598-018-36217-2

Figure Lengend Snippet: Evaluation of direct miRNA-29b binding to the XYLT1 3′UTR. SW1353 cells were seeded and incubated for 24 h. Cells were transfected with XYLT1 3′UTR coding luciferase vectors and miRNA-29b or a negative control miRNA (NC). Cell lysates were harvested after 48 h, and firefly luciferase activity and renilla luciferase activity were determined. Changes in relative luciferase activity resemble the putative interaction of miRNA-29b with the 3′UTR of XYLT1 mRNA. Results are expressed as means +/− SEM for six biological replicates (Mann-Whitney U test).

Article Snippet: Human chondrosarcoma SW1353 cells, purchased from ATCC (Manassas, USA), were seeded in a cell density of 180,000 cells/6-well to analyze the potential direct binding of miRNA-29b to the XYLT1 3′UTR.

Techniques: Binding Assay, Incubation, Transfection, Luciferase, Negative Control, Activity Assay, MANN-WHITNEY

Journal: Scientific Reports

Article Title: microRNA-29b mediates fibrotic induction of human xylosyltransferase-I in human dermal fibroblasts via the Sp1 pathway

doi: 10.1038/s41598-018-36217-2

Figure Lengend Snippet: Quantification of XYLT1 promoter activity after site-directed mutagenesis of Sp1 transcription factor binding sites. The XYLT1 wildtype vector (pGL4.10 XYLT1 c. − 1639 to c. + 1 complete ) was used to delete Sp1 transcription factor binding sites by site-directed mutagenesis. Modified sequences are indicated on the left. SW1353 cells were transfected with the promoter plasmids and relative promoter activity was quantified by dual luciferase assay. Relative promoter activities are referred to the activity of the unmutated wildtype construct, which was defined as 100%. Values are means ± SEM of six biological replicates. **p < 0.01; ***p < 0.001; ****p < 0.0001 (Mann-Whitney U-test).

Article Snippet: Human chondrosarcoma SW1353 cells, purchased from ATCC (Manassas, USA), were seeded in a cell density of 180,000 cells/6-well to analyze the potential direct binding of miRNA-29b to the XYLT1 3′UTR.

Techniques: Activity Assay, Mutagenesis, Binding Assay, Plasmid Preparation, Modification, Transfection, Luciferase, Construct, MANN-WHITNEY

Journal: Osteoarthritis and cartilage

Article Title: Novel cartilage-specific splice variants of fibronectin.

doi: 10.1053/joca.2002.0792

Figure Lengend Snippet: Fig. 2. RT-PCR analysis of bovine articular cartilage FN splice variants in the C-terminal region spanning the III-12 to the I-12 domains. RNA was isolated from bovine metacarpal–phalangeal articular cartilage and two human cell lines, SW1353 and HepG2, cultured in monolayer. RT-PCR was carried out using the VDF and VDR primers as described in Materials and Methods and separ- ated by agarose gel electrophoresis. (A) Lane 1—bovine articular cartilage, Lane 2—SW1353, Lane 3—HepG2, Lane 4—water control. SM—size markers. (B) The cartilage specific RT-PCR products obtained from the bovine articular cartilage sample were recovered and DNA sequenced. The schematic illustrates the FN splice variants identified. Those domains subject to alternative splicing are shown hatched.

Article Snippet: Isolated chondrocytes and the human cell lines, HepG2 (hepatoma ATCC No. HB-8065) and SW1353 (chondrosarcoma ATCC No. HTB-94) were maintained as monolayers in DMEM with 10% (v/v) FCS (Gibco BRL), 100 U/ml penicillin and 100 g/ml streptomycin and passaged after reaching confluence.

Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation, Cell Culture, Agarose Gel Electrophoresis, Control, Alternative Splicing

Journal: Clinical Rheumatology

Article Title: Integrating TWAS, chemical-genomic networks, and experimental models: melatonin as a ferroptosis inhibitor in osteoarthritis therapy

doi: 10.1007/s10067-025-07669-5

Figure Lengend Snippet: A 2D structure of melatonin. B Observation of SW1353 chondrocytes by microscopy. C Effect of different concentrations (0, 5, 10, 20, 40, and 80 μM) of IL-1β on chondrocyte viability. D Effect of different concentrations (0, 5, 10, 20, 40, 80 μM) of melatonin on chondrocyte viability. E Effect of melatonin (0, 5, 10, 20, 40, 80 μM) on the viability of chondrocytes in the presence of IL-1β (10 ng/mL). Significance: ns, p ≥ 0.05; * p < 0.05; ** p < 0.01; p < 0.001

Article Snippet: SW1353 human chondrocytes were purchased from ProCell and cultured in DMEM supplemented with 10% heat-inactivated FBS and 1% penicillin (100 IU/mL) at 5% CO 2 and 37 °C.

Techniques: Microscopy

Journal: Oncology reports

Article Title: Targeting glucose metabolism in chondrosarcoma cells enhances the sensitivity to doxorubicin through the inhibition of lactate dehydrogenase-A.

doi: 10.3892/or.2014.3156

Figure Lengend Snippet: Figure 1. LDHA is upregulated in human chondrosarcoma cells. (A) Western blot analysis of human chondrocyte cell lines CHON-001 and CHON-002 and chondrosarcoma cell lines SW1353 and Hs 819.T. β-actin was a loading control. (B) Western blot analysis of human primary chondrocyte and tumor samples of chondrosarcoma patients. (C) mRNA expression of LDHA in the cell lines as indicated above. mRNA levels of LDHA were increased in chondrosarcoma cells compared with benign chondrocyte. (D) The relative expression of LDHA in chondrosarcoma analyzed from Oncomine.com. (E) Relative activity of LDHA in the above cells. The activities of chondrosarcoma cells were higher than chondrocyte. Columns, mean of three independent experiments; bars, SE.

Article Snippet: Hs 819.T and SW1353 human chondrosarcoma cells were purchased from ATCC.

Techniques: Western Blot, Control, Expressing, Activity Assay

Journal: Oncology reports

Article Title: Targeting glucose metabolism in chondrosarcoma cells enhances the sensitivity to doxorubicin through the inhibition of lactate dehydrogenase-A.

doi: 10.3892/or.2014.3156

Figure Lengend Snippet: Figure 2. Doxorubicin induces LDHA expression and activity. (A) Chondrosarcoma cells Hs 819.T and SW1353 were treated by doxorubicin at 50 and 100 nM for 72 h followed by western blot analysis. (B) Chondrosarcoma cells Hs 819.T and SW1353 were treated by doxorubicin at 50 and 100 nM for 72 h followed by real-time PCR analysis. (C) The activities of LDHA were induced by the indicated doxorubicin treatments for 72 h under regular cell culture conditions. Columns, mean of three independent experiments; bars, SE. **P<0.01. ***P<0.001.

Article Snippet: Hs 819.T and SW1353 human chondrosarcoma cells were purchased from ATCC.

Techniques: Expressing, Activity Assay, Western Blot, Real-time Polymerase Chain Reaction, Cell Culture

Journal: Oncology reports

Article Title: Targeting glucose metabolism in chondrosarcoma cells enhances the sensitivity to doxorubicin through the inhibition of lactate dehydrogenase-A.

doi: 10.3892/or.2014.3156

Figure Lengend Snippet: Figure 3. Doxorubicin-resistant chondrosarcoma cells are sensitive to glucose starvation. (A) Generation of doxorubicin-resistant cells. SW1353 cells were treated with gradually increasing concentrations of doxorubicin in regular cell culture conditions for the selection of resistant cells. SW1353 doxorubicin- resistant clones were pooled and analyzed by the treatments of doxorubicin at indicated concentrations for 72 h and were then analyzed by the cell viability assay. (B) Doxorubicin-resistant cells and parental cells were cultured under glucose depletion medium for 8, 16 and 36 h and were then analyzed by cell viability assay. Columns, mean of three independent experiments; bars, SE. *P<0.05. ***P<0.001.

Article Snippet: Hs 819.T and SW1353 human chondrosarcoma cells were purchased from ATCC.

Techniques: Cell Culture, Selection, Clone Assay, Viability Assay

Journal: Oncology reports

Article Title: Targeting glucose metabolism in chondrosarcoma cells enhances the sensitivity to doxorubicin through the inhibition of lactate dehydrogenase-A.

doi: 10.3892/or.2014.3156

Figure Lengend Snippet: Figure 5. Knockdown of LDHA by siRNA re-sensitizes chondrosarcoma cells to doxorubicin. (A) Knockdown of LDHA by siRNA in SW1353 parental and doxorubicin-resistant cells. β-actin was a loading control. (B) Knockdown of LDHA in both SW1353 parental (left panel) and doxorubicin-resistant (right panel) cells increased the sensitivity to doxorubicin. Columns, mean of three independent experiments; bars, SE.

Article Snippet: Hs 819.T and SW1353 human chondrosarcoma cells were purchased from ATCC.

Techniques: Knockdown, Control

Journal: Oncology reports

Article Title: Targeting glucose metabolism in chondrosarcoma cells enhances the sensitivity to doxorubicin through the inhibition of lactate dehydrogenase-A.

doi: 10.3892/or.2014.3156

Figure Lengend Snippet: Figure 6. The combination of doxorubicin and oxamate shows synergistic inhibitory effects on chondrosarcoma cells. (A) SW1353 parental (left) and doxoru bicin-resistant (right) cells were treated with doxorubicin alone, oxamate alone and combination of doxorubicin with oxamate at the indicated concentration for 72 h followed by the cell viability analysis. (B) Colony formation assay analysis of SW1353 parental (left panel) and resistant (right panel) cells. Five hundred cells of each experiment were seeded on a 10-cm dish with indicated drugs in regular cell culture medium. The medium and drugs were refreshed every two days. Cells were grown for 3 weeks and the surviving colonies were stained with gentian violet after methanol fixation, and visible colonies (>50 cells) were counted. Colonies from randomly-selected image areas of three replicate wells were enumerated. Columns, mean of three independent experiments; bars, SE. *P<0.05. **P<0.01.

Article Snippet: Hs 819.T and SW1353 human chondrosarcoma cells were purchased from ATCC.

Techniques: Concentration Assay, Colony Assay, Cell Culture, Staining

Journal: Oncology reports

Article Title: Targeting glucose metabolism in chondrosarcoma cells enhances the sensitivity to doxorubicin through the inhibition of lactate dehydrogenase-A.

doi: 10.3892/or.2014.3156

Figure Lengend Snippet: Figure 7. In vivo experiments demonstrate the combination of doxorubicin and glycolysis inhibitors has synergistic effects on the inhibition of chondrosarcoma growth. (A) Mice were inoculated SW1353 parental cells and (B) SW1353 doxorubicin-resistant cells. Once the tumor established, mice were treated with control, doxorubicin alone, oxamate alone and doxorubicin plus oxamate for eight consecutive weeks.

Article Snippet: Hs 819.T and SW1353 human chondrosarcoma cells were purchased from ATCC.

Techniques: In Vivo, Inhibition, Control

Journal: Arthritis Research & Therapy

Article Title: Chondrocytes, synoviocytes and dermal fibroblasts all express PH-20, a hyaluronidase active at neutral pH

doi: 10.1186/ar1730

Figure Lengend Snippet: Northern blot analysis of mRNA from various connective tissue cell lines. (a) A radiolabelled PH-20 cDNA probe was hybridized to a northern blot containing 25 μg/lane of total RNA from (1) testis (positive control); (2) liver (negative control); (3) synoviocytes; (4) chondrocytes; (5) chondrocytes stimulated with 5 ng/ml of IL-1; (6) chondrocytes stimulated with 10 ng/ml of IL-1; (7) fibroblasts; (8) a chondrosarcoma cell line. (b) For loading control, the same blot was stripped and hybridized with a β-actin probe. (c) Column box–whisker plot showing the relative increase of the PH-20 2.4-kilobase transcript in chondrocytes stimulated with IL-1 at two concentrations ( n = 5; * P = 0.0004 by paired t -test).

Article Snippet: Human chondrosarcoma cells (SW 1353) were purchased from the American Tissue Culture Collection (LGC Promochem, Teddington, UK).

Techniques: Northern Blot, Positive Control, Negative Control, Control, Whisker Assay

Journal: International journal of molecular sciences

Article Title: Targeting Bone Tumours with 45S5 Bioactive Glass.

doi: 10.3390/ijms251910830

Figure Lengend Snippet: Figure 7. 45S5-BG inhibits tumour formation in vivo. OS, GCTB and CS cells were co-transplanted with 45S5-BG at the indicated concentrations onto the CAM of fertilised chicken eggs (n = 20 for each experimental group). Tumours that formed after seven days were resected, and (A) the tumour take rate, (B) the volume of the individual tumours (white rectangles indicate the mean tumour volumes), and (C) the cumulative tumour volume of the experimental groups were calculated. (D) Photographs of the resected tumours. Representative data from the OS cell line MNNG-HOS, the CS cell line SW1353, and a primary GCTB cell line are shown.

Article Snippet: The following five commercially available human osteosarcoma cell lines and two chondrosarcoma cell lines were used in this study: HOS 143B (Sigma-Aldrich, Taufkirchen, Germany), CAL-72 (DSMZ, Braunschweig, Germany), MNNG-HOS, Saos-2, and U2OS (Cell Line Service GmbH, Eppelheim, Germany), SW1353 (Cell Line Service GmbH), and CAL-78 (DSMZ).

Techniques: In Vivo