SW1353 Human Chondrosarcoma Cells Research Articles

MiR-6839-5p inhibits cell proliferation, migration and invasion; a possible correlation with the suppressing VEGFA expression in human chondrosarcoma cells

MicroRNAs play an important role in the proliferation, invasion, and metastasis of malignancy. In previous studies (detailed in our previous paper), the expression of miR-6839-5p was significantly increased in SW1353 cells after 125I seed 6 Gy irradiation, which indicated miR-6839-5p may play a tumor suppression function in chondrosarcoma cells. This study aimed to identify the effects of miR-6839-5p on the human chondrosarcoma cells, and investigate the potential target genes of miR-6839-5p. Firstly, chondrosarcoma cells (SW1353 and CAL78) were transfected with hsa-miR-6839-5p specific mimic. Secondly, Cell viability assay (MTT assay), Colony formation assay, Wound healing assay, Transwell assay, TUNEL staining and Western blotting experiments were performed, and the results proved miR-6839-5p can inhibit chondrosarcoma cells proliferation, migration and invasion. Meanwhile, miR-6839-5p significantly down-regulated apoptosis facilitator Bcl-2 expression, and promoted apoptosis of chondrosarcoma cells. It is reasonable to speculate miR-6839-5p might downregulate Bcl-2 expression to induce apoptosis in SW1353 human chondrosarcoma cells. Lastly, RNA extraction and bioinformatic analysis was performed on SW1353 cells transfected with hsa-miR-6839-5p specific mimic to investigate the potential target genes of miR-6839-5p. A total of 253 differentially expressed mRNA genes (105 up-regulated genes and 148 down-regulated genes) were found, and 23 differentially expressed downregulated genes were identified. Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to validate the results, which demonstrated the expression of BST2, VEGFA, FPR3 and PPARA was significantly downregulated by miR-6839-5p mimic. Furthermore, miR-6839-5p inhibitor can restore or partially restore the expression value of the above four genes. The analysis results of miRNA target gene prediction database indicated VEGFA was the most likely direct target gene of miR-6839-5p.

Transcriptomic Analysis of the Anticancer Effects of Annatto Tocotrienol, Delta-Tocotrienol and Gamma-Tocotrienol on Chondrosarcoma Cells.

Previous studies have demonstrated the anticancer activities of tocotrienol on several types of cancer, but its effects on chondrosarcoma have never been investigated. Therefore, this study aims to determine the anticancer properties of annatto tocotrienol (AnTT), γ-tocotrienol (γ-T3) and δ-tocotrienol (δ-T3) on human chondrosarcoma SW1353 cells. Firstly, the MTT assay was performed to determine the half-maximal inhibitory concentration (IC50) of tocotrienol on SW1353 cells after 24 h treatment. The mode of cell death, cell cycle analysis and microscopic observation of tocotrienol-treated SW1353 cells were then conducted according to the respective IC50 values. Subsequently, RNAs were isolated from tocotrienol-treated cells and subjected to RNA sequencing and transcriptomic analysis. Differentially expressed genes were identified and then verified with a quantitative PCR. The current study demonstrated that AnTT, γ-T3 and δ-T3 induced G1 arrest on SW1353 cells in the early phase of treatment (24 h) which progressed to apoptosis upon 48 h of treatment. Furthermore, tocotrienol-treated SW1353 cells also demonstrated large cytoplasmic vacuolation. The subsequent transcriptomic analysis revealed upregulated signalling pathways in endoplasmic reticulum stress, unfolded protein response, autophagy and transcription upon tocotrienol treatment. In addition, several cell proliferation and cancer-related pathways, such as Hippo signalling pathway and Wnt signalling pathway were also significantly downregulated upon treatment. In conclusion, AnTT, γ-T3 and δ-T3 possess promising anticancer properties against chondrosarcoma cells and further study is required to confirm their effectiveness as adjuvant therapy for chondrosarcoma.

An Anti-Inflammatory Composition of Boswellia serrata Resin Extracts Alleviates Pain and Protects Cartilage in Monoiodoacetate-Induced Osteoarthritis in Rats.

The boswellic acids, the active compounds in Boswellia serrata gum resin extract, are potent anti-inflammatory agents and are specific nonredox inhibitors of 5-Lipoxygenase (5-LOX). Here, we present the anti-osteoarthritis (OA) efficacy of LI13019F1 (also known as Serratrin®), a unique composition containing the acidic and nonacidic fractions of B. serrata gum resin. This composition strongly inhibited 5-LOX activity with the half-maximal inhibitory concentration (IC50) of 43.35 ± 4.90 μg/mL. Also, LI13019F1 strongly inhibited the leukotriene B4 (IC50, 7.80 ± 2.40 μg/mL) and prostaglandin E2 (IC50, 6.19 ± 0.52 μg/mL) productions in human blood-derived cells. Besides, LI13019F1 reduced TNF-α production with the IC50 of 12.38 ± 0.423 μg/mL. On average, 1, 2.5, and 5 μg/mL doses of LI13019F1 protected 34.62, 47.66, and 62.29% SW1353 human chondrosarcoma cells from IL-1β induced SOX-9 depletion, respectively. Further, a 28-day preclinical proof-of-concept study evaluated the pain relief efficacy of LI13019F1 in monoiodoacetate- (MIA-) induced Sprague-Dawley rats. At the end of the study, 150 and 300 mg/kg doses of LI13019F1 supplemented rats showed significant improvements (55.17 ± 5.81 g (p < 0.05), and 66.22 ± 6.30 g (p < 0.05), respectively, vs. MIA: 31.22 ± 7.15 g) in body-weight-bearing capacities. Concurrently, LI13019F1-150 and LI13019F1-300 rats substantially (p < 0.05) increased the threshold of pain sensitivity to pressure (26.98 ± 2.36 and 28.06 ± 2.72-gram force, respectively; vs. 18.63 ± 5.82 in MIA) and increased (p < 0.05) the latent time to withdraw the paw after a thermal stimulus (23.61 ± 2.73 and 28.18 ± 1.90 sec, respectively; vs. 16.56 ± 1.22 sec. in MIA). Besides, the histological observations on Safranin-O green stained articular cartilage revealed that LI13019F1 also prevented the MIA-induced structural damage of the cartilage and reduced the loss of the extracellular matrix (ECM) components in the experimental rats. In conclusion, the present observations suggest that LI13019F1, a new composition of B. serrata gum resin extracts, reduces pain and protects articular cartilage from the damaging action of MIA in a rodent model.

Analysis of Cells Proliferation and MicroRNAs Expression Profile in Human Chondrosarcoma SW1353 Cells Exposed to Iodine-125 Seeds Irradiation

Chondrosarcoma is the second most common bone malignancy in adults, and it is often resistant to traditional chemotherapy and radiation therapy. Permanent implantation of iodine-125 (125I) seeds has been explored for the treatment of many types of cancer. In this study, the aim was to investigate the proliferative and microRNA (miRNA) effects of 125I seeds irradiation on human chondrosarcoma SW1353 cells. First, a new in vitro 125I seed irradiation model was established, and cell viability and miRNA microarray assays were performed before and after exposure to the 125I seeds. Cell proliferation was inhibited, and miRNA expression was substantially altered by irradiation exposure. The inhibition of cell proliferation was positively correlated with increased radiation doses, with cells showing the highest total radiation dose 7 days after irradiation. A total of 2549 miRNAs were detected in the SW1353 cells after exposure to 6 Gy of radiation, which included 189 differentially expressed miRNAs (98 upregulated and 91 downregulated). Four miRNAs were found to play important roles in the inhibition of cell proliferation after irradiation exposure, including miR-1224-5p, miR-492, miR-135b-5p, and miR-6839-5p. The target genes of the associated miRNAs mentioned were vascular endothelial growth factor A (VEGFA), C-X-C motif chemokine 12 (CXCL12), mitogen-activated protein kinase kinase kinase kinase 3 (MAP4K3), and apoptosis facilitator Bcl-2-like protein 14 (BCL2L14). Hence, the mitogen-activated protein kinase signaling pathway may be involved in how chondrosarcoma cells respond to 125I seed irradiation.

Effects of neuraminidase 3 gene over-expression on proliferation and migration of chondrosarcoma cells

Objective To observe the neuraminidase 3 (NEU3) expression and location in SW1353 cells and investigate the effect of NEU3 expression on the proliferation and migration of human chondrosarcoma SW1353 cells. Methods The NEU3 overexpression cell line was constructed by the lipo 3000 transfection reagent. The experiment was divided into 4 groups: UN, and no treatment was given. Blank, was added with equal dose transfection reagent.pcon, was added with no-load plasmid. pNEU3, NEU3 gene overexpression plasmid was added. Immunofluorescence was used to determine the expression of NEU3 in SW1353 cells. Different groups of cells were added the lipo3000 transfection reagent and to explore these groups' mRNA expression of NEU3 by Real-time polymerase chain reaction (PCR). The impact of NEU3 over-expression on the migration of SW1353 cells was examined by scratch test. The effect of NEU3 over-expression on the proliferation of SW1353 cells was measured by EdU proliferation test and CCK-8 assay. The difference in the NEU3 expression and cell capability was tested by two independent samples t test. Results After transfection with NEU3 gene over-expressed plasmid, Real-time PCR showed that NEU3 gene mRNA was significantly up-regulated. EdU proliferation test showed that NEU3 gene over-expression could promote proliferation of SW1353 proliferation cells [positive rate for control group: (6.56±2.29)%, (11.81±6.49)%, (8.64±3.90)%; for treatment group: (40.57±14.32)% (t=11.355, P<0.05]. The scratch test showed that NEU3 gene over-expression could enhance the migration of SW1353 cells [Remian Scratch area for control group: (18 343±367), (20 803±124), (26 472±483) dpi; for treatment group: (3 320±193) dpi, t=7.265, P<0.05]. Conclusion Over-expression of NEU3 gene can enhance proliferation and migration of human chondrosarcoma SW1353 cells. Key words: Chondrosarcoma; Sialidase; Cell proliferation; Cell migration

Activation of G-protein-coupled bile acid receptor Gpbar1 (TGR5) inhibits degradation of type II collagen and aggrecan in human chondrocytes

Abnormal loss of components of the extracellular matrix (ECM) including type II collagen and aggrecan caused by proinflammatory cytokines such as tumor necrosis factor-α (TNF-α) is an important pathophysiological characteristic of osteoarthritis (OA). G-protein-coupled bile acid receptor, Gpbar1 (TGR5), is an important member of the bile acid receptor subclass of G Protein-Coupled Receptors (GPCRs). Little information regarding the effects of TGR5 in the pathological development of OA has been reported before. In the current study, we showed that TGR5 is expressed in human primary chondrocytes and human chondrosarcoma SW1353 cells. Interestingly, expression of TGR5 was reduced in response to TNF-α treatment in SW1353 cells. Our results indicate that activation of TGR5 using its specific agonist INT-777 reduced TNF-α-induced degradation of the articular ECM, including type II collagen and aggrecan, by inhibiting expression of matrix metalloproteinase-3 (MMP-3), MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs- 4 (ADAMTS-4) and ADAMTS-5. We also found that INT-777 treatment inhibited phosphorylation of p38 and activation of the IκB kinase/inhibitory κBα/nuclear factor- κB (IKK/IκBα/NF-κB) signaling pathway. Notably, knockdown of TGR5 abolished the protective effects of INT-777 against ECM degradation, suggesting the involvement of TGR5. Our findings implicate that TGR5 might be considered as a potential therapeutic target for the treatment of OA.

Pyrroloquinoline quinone induces chondrosarcoma cell apoptosis by increasing intracellular reactive oxygen species.

Pyrroloquinoline quinone (PQQ) has been reported to contribute to cancer cell apoptosis and death; however, little is known of its underlying mechanisms. The present study was designed to investigate the role of PQQ in chondrosarcoma cell apoptosis and the underlying mechanism. A cell cytotoxicity assay was used to detect cell death; flow cytometry analysis was also performed to determine cell apoptosis and intracellular reactive oxygen species (ROS). Biochemical methods were employed to detect the activity and the expression of superoxide dismutase (SOD)1, SOD2 and glutathione. The present study also examined the effect on tumorigenesis in vivo. The results demonstrated that the apoptosis of SW1353 cells induced by PQQ increased in a concentration- and time-dependent manner, which may be attributable to the accumulation of intracellular ROS. In the in vivo experiments, PQQ inhibited proliferation and promoted apoptosis, increased ROS levels and caused DNA damage in transplanted cells. Taken together, the findings of the present study confirmed that PQQ induced apoptosis in human chondrosarcoma SW1353 cells and transplanted cells, by increasing intracellular ROS and reducing the ability of scavenging oxygen free radicals.

MicroRNA-17-5p contributes to osteoarthritis progression by binding p62/SQSTM1.

Autophagy has been reported to be widely involved in the pathogenesis of osteoarthritis (OA). Increasing evidence suggested the important role of microRNAs (miRs) in the progression of OA. However, the functional role of miR-17-5p in OA development has remained to be fully elucidated. First, a mouse model of OA was established and the relative level of miR-17-5p was determined using PCR. Safranin O-fast green staining was applied to determine cartilage degeneration. TargetScan software and a dual luciferase reporter assay were applied to determine potential target genes of miR-17-5P. Autophagy measurement was performed using green fluorescent protein-microtubule-associated protein 1 light chain 3 (LC3) dot analysis. The results demonstrated that the relative expression of miR-17-5p was significantly decreased in OA model mice. In addition, the level of miR-17-5p was decreased in SW1353 human chondrosarcoma cells treated with interleukin-1β. Furthermore, autophagy was found to be suppressed in the knee joints of experimental OA model mice. The dual luciferase reporter assay confirmed that p62/sequestosome 1 was a target gene of miR-17-5p. Of note, miR-17-5p inhibitor-induced reduction of LC3 dots was markedly reversed by knockdown of p62 in SW1353 cells. In conclusion, decreased miR-17-5p expression in chondrocytes induced autophagy mainly through suppressing the expression of p62, thereby contributing to OA progression.

Puerarin induces cell apoptosis in human chondrosarcoma cell line SW1353 via inhibition of the PI3K/Akt signaling pathway

Chondrosarcoma is a malignant soft tissue sarcoma with poor prognosis. Puerarin has been demonstrated to possess anticancer properties; however, the effects of puerarin in human chondrosarcoma cells remain unknown. The present study aimed to investigate the anticancer effects of puerarin in SW1353 human chondrosarcoma cells. SW1353 cells were treated with increasing concentrations of puerarin for different durations. Cell viability was evaluated using MTT assays. Cell apoptosis rates were determined by flow cytometry. The activities of caspase-3 and caspase-9 were measured by enzymatic assay. The expression of RAC-alpha serine/threonine-protein kinase (Akt), phosphorylated-Akt, caspase-3 and apoptosis-associated proteins, including B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) were detected by western blotting. Puerarin significantly decreased cell viability and significantly induced apoptosis of SW1353 cells. In addition, puerarin significantly increased the enzymatic activities of caspase-3 and caspase-9. Puerarin treatment suppressed the expression of p-Akt and Bcl-2 but promoted the expression of Bax and cleaved caspase-3 in SW1353 cells. Notably, the phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 abrogated the decreased phosphorylation of Akt, suggesting that the PI3K/Akt signaling pathway is involved in mediating the anticancer effects of puerarin. The data from the present study indicated that puerarin exhibits anticancer effects in SW1353 cells and may be a potential therapeutic drug for patients with chondrosarcoma.

Increased receptor activator of nuclear factor κβ ligand/osteoprotegerin ratio exacerbates cartilage destruction in osteoarthritis in vitro.

Osteoarthritis (OA) is a degenerative joint disease characterized by progressive cartilage destruction, matrix degradation and bony changes. Subchondral bone alterations in osteoarthritis are associated with cartilage destruction. It has previously been demonstrated that osteoprotegerin (OPG) and receptor activator of nuclear factor κβ ligand (RANKL) mediate this process. The RANKL/OPG ratio is altered in OA chondrocytes compared with normal chondrocytes. In the pathogenesis of OA, abnormal expression levels of matrix metalloproteinase-13 (MMP-13) are secreted by chondrocytes has a vital role in the progression of cartilage erosion. In the present study, the effect of various RANKL/OPG ratios on MMP-13 expression levels was investigated in interleukin-1β-stimulated SW1353 human chondrosarcoma cells. Cell viability was assessed by MTT assay and MMP-13 mRNA and protein expression levels were analyzed by quantitative reverse-transcription-quantitative polymerase chain reaction, ELISA and western blot analyses, respectively. The results demonstrated that an increase in MMP-13 mRNA and protein expression levels was observed with increasing RANKL/OPG ratio. These findings suggest that this mechanism may be used as a novel therapeutic strategy against OA.

Docosahexenoic acid treatment ameliorates cartilage degeneration via a p38 MAPK-dependent mechanism.

Osteoarthritis (OA) is a common chronic inflammatory disease, characterized by cartilage degradation. The aberrant expression of matrix metalloproteinase-13 (MMP-13) plays a vital role in the pathogenesis of OA. The anti-inflammatory property of docosahexenoic acid (DHA) was previously revealed and showed that DHA retards the progress of many types of inflammatory disease. To evaluate the prophylactic function of DHA in OA, the effect of DHA on cartilage degeneration was assessed in interleukin-1β (IL-1β) stimulated human chondrosarcoma SW1353 cells or a rat model of adjuvant-induced arthritis (AIA). The safe concentration range (0–50 µg/ml in vitro) of DHA was determined by flow cytometry and MTT assay. The inhibitory effects of DHA on MMP-13 mRNA and protein expression were confirmed by RT-qPCR, ELISA and western blotting. Furthermore, findings of an in vivo study showed that DHA can increase the thickness of articular cartilage and decrease MMP-13 expression in cartilage matrix in a rat AIA model. We also revealed the mechanism by which DHA ameliorates cartilage degeneration from OA. The DHA-mediated inhibition of MMP-13 expression was partially attributed to the inactivation of the p38 mitogen-activated protein kinases pathway by suppressing p-p38 in IL-1β-stimulated SW1353 cells and a rat AIA model. Our findings suggested that DHA is a promising therapeutic agent that may be used for the prevention and treatment of OA.

Alterative effects of an oral alginate extract on experimental rabbit osteoarthritis.

BackgroundOsteoarthritis (OA) is a common joint disease that causes disabilities in elderly. However, few agents with high efficacy and low side effects have been developed to treat OA. In this study, we evaluated the effects of the alginate extract named CTX in OA cell and rabbit models.ResultsCTX was formulated by hydrolyzing sodium alginate polymers with alginate lyase and then mixing with pectin. HPLC was used to analyze the CTX content. Human chondrosarcoma SW1353 cells treated with interleukin-1β were used as OA model cells to investigate the effects of CTX on chondrocyte inflammation and anabolism. CTX at concentrations up to 1000 μg/ml exerted low cytotoxicity. It inhibited the gene expression of proinflammatory matrix metalloproteinases (MMPs) including MMP1, MMP3 and MMP13 in a dose-dependent manner and increased the mRNA level of aggrecan, the major proteoglycan in articular cartilage, at 1000 μg/ml. Thirteen-week-old New Zealand White rabbits underwent a surgical anterior cruciate ligament transection and were orally treated with normal saline, glucosamine or CTX for up to 7 weeks. Examinations of the rabbit femur and tibia samples demonstrated that the rabbits taking oral CTX at a dosage of 30 mg/kg/day suffered lesser degrees of articular stiffness and histological cartilage damage than the control rabbits.ConclusionsThe gene expression profiles in the cell and the examinations done on the rabbit cartilage suggest that the alginate extract CTX is a pharmaco-therapeutic agent applicable for OA therapy.

Gallic acid induces apoptosis and inhibits cell migration by upregulating miR-518b in SW1353 human chondrosarcoma cells

Gallic acid (GA), a natural agent, is widely distri-buted in plants with a range of biological effects and has been of potential interest as anticancer agent. However, its effects on chondrosarcoma cell apoptosis are still undefined. In the present study, the possible mechanisms of GA-induced apoptosis were explored in SW1353 cells, a human chondrosarcoma cell line. Our results showed that GA inhibited cell viability dose- and time-dependently. Morphological examination of GA-treated cells exhibited the typical features of cell death, such as rounding up of the cells and cell shrinkage. Wound-healing assay indicated that GA inhibited the migratory abilities of SW1353 cells. Hoechst 33258 staining assay and Annexin V/PI staining assay exhibited apoptosis induction by GA. To determine the molecular mechanism of GA-induced apoptosis, the expression levels Bcl-2, Bax, caspase-3 and caspase-9 were determined in SW1353 cells treated with GA. We found that GA downregulated the expression of the anti-apoptotic protein Bcl-2, and upregulated the expression of the pro-apoptotic protein Bax, and the activation of caspase-3 and caspase-9. To identify the possible mechanisms, the changes of microRNA expression were tested using the miRCURY™ LNA expression array. It was observed that the miR-518b gene was upregulated in treated cells. Taken together, these data show that GA induces apoptosis and inhibits cell migration by upregulating miR-518b in SW1353 cells.

Sclerostin regulation of WISP1 and MMPS in chondrocytes is dependent on the LRP5/6 binding domain but not secretion from the cell

Sclerostin regulation of WISP1 and MMPS in chondrocytes is dependent on the LRP5/6 binding domain but not secretion from the cell

Chemical constituents of volatile oil from Pyrolae herba and antiproliferative activity against SW1353 human chondrosarcoma cells

The objective of the present study was to identify chemical constituents of volatile oil from Pyrolae herba (PHVO) and evaluate the antiproliferative activity of PHVO against SW1353 human chondrosarcoma cells. The volatile oil from Pyrolae herba was prepared by hydrodistillation and characterized by gas chromatography-mass spectroscopy (GC-MS). A total of 12 components in PHVO were identified representing 81.62% of the total integrated chromatographic peaks. The major compounds were found to be n-hexadecanoic acid (29.29%), cedrol (17.08%), 6,10,14-trimethyl-2-pentadecanone (9.59%) and cis-9-octadecadienoic acid (8.23%). The antiproliferative activity of PHVO against SW1353 cells was investigated using MTT assay, flow cytometry and western blot analysis. Our results demonstrated that PHVO inhibited SW1353 cell viability in a dose- and time-dependent manner. Furthermore, PHVO treatment decreased the number of cells entering the S phase and caused a reduction in the expression of cyclin D1, cyclin-dependent kinase (CDK)4 and CDK6, whereas it caused an increase in the expression of p21. PHVO demonstrated potent antitumor activity against SW1353 cells, suggesting its potential use as a therapeutic agent in the treatment of chondrosarcoma.

Astaxanthin Mediates Inflammation Biomarkers Associated with Arthritis in Human Chondrosarcoma Cells Induced with Interleukin-1β

Astaxanthin, a potent antioxidant, may downregulate inflammation-associated factors involved in arthritis. SW-1353 human chondrosarcoma cells were pre-incubated with 0, 0.01, 0.1, or 1.0 μmol/L astaxanthin for 48 h, and oxidative stress induced with 10 ng/mL IL-1β overnight. Astaxanthin (1.0 μmol/L) accumulated in SW1353 cells in a time-dependent manner. IL-1β alone suppressed intracellular antioxidant activity, resulting in overproduction of reactive oxygen species (ROS), matrix metalloproteinase (MMP)-13, inflammatory cytokines and mediators. In contrast, pre-incubation with astaxanthin increased (P < 0.05) glutathione peroxidase activity and decreased (P < 0.05) ROS, MMP-13, IL-6, TNF-α and inflammatory mediators. Pretreatment with astaxanthin also downregulated transcriptional activation of nuclear factor-κB and activator protein-1, which play critical roles in downstream production of MMP, inflammatory cytokines and mediators. Astaxanthin protects against degenerative factors upregulated by IL-1β, likely by scavenging ROS required for transcriptional activation.

Cryptoxanthin and lutein Mediate Inflammation in Human Chondrosarcoma Cells through Nuclear Factor-B and Activator Protein-1

-Cryptoxanthin (Cr) and lutein (Lu) may downregulate inflammation-associated factors involved in arthritis. The uptake and protective effects of Cr and Lu, alone and in combination, against chondrocyte dysfunction were examined using SW-1353 human chondrosarcoma cells stress-induced with IL-1β. Cr and Lu (10.0 mol/L) accumulated in SW-1353 cells in a time-dependent manner; Cr and Lu in combination inhibited Cr uptake. Cells were pre-incubated with 0, 0.01, 0.1, or 1.0 mol/L Cr, Lu, or a mixture of Cr and Lu for 48 h, then oxidative stress induced with 10 ng/mL IL-1 for 18 h. Both Cr and Lu alone and in combination increased (P < 0.05) glutathione peroxidase activity and decreased intracellular reactive oxygen species. Cr (0.1 mol/L) decreased (P < 0.05) matrix metalloproteinase (MMP)-13, TNF-, and inflammatory mediator production, as well as nuclear factor-B (NF-B) and activator protein-1 (AP-1) activity, at the expense of IL-2 and IFN- upregulation. Lu decreased (P < 0.05) TNF-, inflammatory mediators and NF-B, but not MMP-13 or AP-1. In combination, Cr and Lu decreased (P < 0.05) MMP-13, inflammatory mediators, NF-B and AP-1, without inflammatory cytokine upregulation. Cr and Lu protect against degenerative factors upregulated by IL-1 by scavenging reactive oxygen species required for transcriptional activation.

Lutein and β-Cryptoxanthin Inhibit Inflammatory Mediators in Human Chondrosarcoma Cells Induced with IL-1β

Objective: Studies have shown that lutein (Lu) and β-cryptoxanthin (βCr) may down-regulate factors involved in inflammation associated with osteoarthritis and rheumatoid arthritis. We studied the possible protective effects of Lu and βCr in vitro against human chondrocyte dysfunction using a human chondrosarcoma cell line. Methods: SW-1353 human chondrosarcoma cells were cultured for 24 hr in supplemented medium containing 0, 0.01, 0.1 or 1.0 µmol/L of Lu or βCr and subsequently stressed for 24 hr in the presence of 10 µg/L IL-1β. The resulting condi- tioned medium was analyzed for matrix-metalloproteinase-13 (MMP-13), cytokines (IL-1α, IL-2, IL-4, IL-10, IFN-γ , IL- 6, IL-8, and TNF-α), and PGE2. Nuclear extract from the harvested cells was analyzed for NFκB. Results: Lu (1.0 µmol/L; P<0.05) but not βCr decreased MMP-13. Both Lu (1.0 µmol/L; P<0.05) and βCr (0.1 and 0.01 µmol/L; P<0.01) inhibited PGE 2 production. All concentrations of βCr suppressed (P<0.05) IL-1α, IL-2 and IFN-γ pro- duction while Lu increased concentrations of these cytokines. Lu increased (P<0.05) while βCr decreased IL-4 and IL-10 concentrations. NFκB p50 production was suppressed (P<0.01) by both Lu and βCr, with Lu being more inhibitory. Conclusion: Therefore, Lu and βCr protected against IL-1β-induced chondrocyte dysfunction by down-regulating NFκB activation and inhibiting inflammatory response, albeit through somewhat different pathways.

Millimeter wave radiation induces apoptosis via affecting the ratio of Bax/Bcl-2 in SW1353 human chondrosarcoma cells

The efficacy and safety of millimeter wave radiation has been proven for various types of malignant tumors. However, the mechanisms underlying effects of millimeter wave radiation on apoptosis are still unclear. The present study was undertaken to examine the effects of millimeter wave radiation on cell apoptosis and mitochondrial membrane potential, and to determine the molecular mechanism of millimeter wave radiation-induced apoptosis by investigating the expression of Bcl-2 family proteins (Bcl-2, Bax), caspase-9 and caspase-3 in SW1353 cells. We found that millimeter wave radiation suppressed the viability of SW1353 cells, demonstrating that millimeter wave radiation induced cell apoptosis and reduced cell viability in a time-dependent manner. Furthermore, we observed that treatment of cells with millimeter wave radiation significantly induced loss of mitochondrial membrane potential, upregulated proapoptotic Bax, caspase-9 and caspase-3, but did not significantly change levels of antiapoptotic Bcl-2. These data suggested that millimeter wave radiation may induce apoptosis via affecting the ratio of Bax/Bcl-2 in SW1353 cells.

α-Mangostin Induces Apoptosis in Human Chondrosarcoma Cells through Downregulation of ERK/JNK and Akt Signaling Pathway

Chondrosarcoma is a malignant primary bone tumor that is resistant to chemotherapy and radiation therapy. α-Mangostin, a component of Garcinia mangostana Linn, is a xanthone derivative shown to have antioxidant and antitumor properties. This study is the first to investigate anticancer effects of α-mangostin in the human chondrosarcoma cell line SW1353. We showed that α-mangostin inhibited cell proliferation of SW1353 cells in a time- and dose-dependent manner by using the trypan blue exclusion method. Hoechst 33342 nuclear staining and nucleosomal DNA-gel electrophoresis revealed that α-mangostin could induce nuclear condensation and fragmentation, typically seen in apoptosis. Flow cytometry using Annexin V/PI double staining assessed apoptosis, necrosis and viability. α-Mangostin activated caspase-3, -8, -9 expression, decreased Bcl-2 and increased Bax. This promotes mitochondrial dysfunction, leading to the release of cytochrome c from the mitochondria to the cytoplasm. In addition, total and phosphorylated ERK and JNK were downregulated in α-mangostin-treated SW1353 cells but no changes in p38. α-Mangostin also decreased phosphorylated Akt without altering total Akt. These results suggest that α-mangostin inhinbited cell proliferation and induced apoptosis through downregulation of ERK, JNK and Akt signaling pathway in human chondrosarcoma SW1353 cells.