AMP-activated protein kinase activation mediates CCL3-induced cell migration and matrix metalloproteinase-2 expression in human chondrosarcoma
© Hsu et al.; licensee BioMed Central Ltd. 2013
Received: 17 April 2013
Accepted: 3 September 2013
Published: 18 September 2013
Chemokine (C-C motif) ligand 3 (CCL3), also known as macrophage inflammatory protein-1α, is a cytokine involved in inflammation and activation of polymorphonuclear leukocytes. CCL3 has been detected in infiltrating cells and tumor cells. Chondrosarcoma is a highly malignant tumor that causes distant metastasis. However, the effect of CCL3 on human chondrosarcoma metastasis is still unknown. Here, we found that CCL3 increased cellular migration and expression of matrix metalloproteinase (MMP)-2 in human chondrosarcoma cells. Pre-treatment of cells with the MMP-2 inhibitor or transfection with MMP-2 specific siRNA abolished CCL3-induced cell migration. CCL3 has been reported to exert its effects through activation of its specific receptor, CC chemokine receptor 5 (CCR5). The CCR5 and AMP-activated protein kinase (AMPK) inhibitor or siRNA also attenuated CCL3-upregulated cell motility and MMP-2 expression. CCL3-induced expression of MMP-2 and migration were also inhibited by specific inhibitors, and inactive mutants of AMPK, p38 mitogen activated protein kinase (p38 or p38-MAPK), and nuclear factor κB (NF-κB) cascades. On the other hand, CCL3 treatment demonstrably activated AMPK, p38, and NF-κB signaling pathways. Furthermore, the expression levels of CCL3, CCR5, and MMP-2 were correlated in human chondrosarcoma specimens. Taken together, our results indicate that CCL3 enhances the migratory ability of human chondrosarcoma cells by increasing MMP-2 expression via the CCR5, AMPK, p38, and NF-κB pathways.
Chondrosarcomas are the third most common bone tumors, after myelomas and osteosarcomas. Chondrosarcomas are rapidly progressive, pathologically diverse, and highly malignant, and to date, surgical resection remains the primary treatment for these sarcomas. They also have the potential for distant metastasis . Therefore, better strategies of treatment will ultimately require understanding of the molecular mechanisms of the metastasis step of human chondrosarcoma and identifying and specifically targeting.
Metastasis is a multistage process that requires cancer cells to escape from the primary tumor, survive in the circulation, seed at distant sites, and grow. Metastasis increases cell motility, induction of vascular and lymphatic angiogenesis, and migration and invasion to other organs . The invasion of tumor cells is a complex, multistage process. To facilitate cell motility, invading cells need to change their cell-cell adhesion properties, rearrange the extracellular matrix (ECM) environment, suppress anoikis, and reorganize their cytoskeleton . MMPs play important roles in these processes because their proteolytic activities assist in the degradation of the ECM and basement membranes [4, 5]. In addition to MMPs, cytokines, growth factors, and chemokines have all been shown to regulate tumor cell invasion through autocrine or paracrine pathways . Previous studies have demonstrated the expression of MMP-1, MMP-2, MMP-3, MMP-9, and MMP-13 in human chondrosarcoma cells . Among the all MMP enzymes, MMP-2 (collagenase-2) is particularly interesting because of its role in cartilage degradation via the breakdown of type II collagen, the major collagen component of cartilage. It has also been reported that MMP-2 plays a critical role in ECM turnover and cell-cell interactions as well as metastases of chondrosarcomas .
Chemokines are low-molecular weight secretory proteins that can regulate the chemotaxis and the metabolic activity of specific leukocyte subsets. Their production, in general, is stimulated by pro-inflammatory cytokines, growth factors, and by pathogenic stimuli arising in inflammatory tissues. In diseased tissues, different tumor cell types trigger different complex chemokine networks that influence the quality and quantity of immune-cell infiltration and, consequently, the proliferation, survival, spread, and angiogenic response of malignant cells . CCL3, also known as macrophage inflammatory protein 1α (MIP-1α), is a pro-inflammatory cytokine belonging to the CC chemokine subfamily and is a ligand for CCR5, which stimulates chemotactic activities in a variety of immune cells such as monocytes, lymphocytes, macrophages [10, 11]. CCL3 has also been implicated in the regulation of cancer cell growth, angiogenesis and metastasis of different tumors such as melanoma , colorectal cancer , and renal cell carcinoma .
Epidemiological studies have shown that energy availability is associated with an increased risk of several metabolic diseases such as obesity, hypertension, diabetes, and induced cancers [15, 16]. Aberrant energy metabolism may induce systemic and chronic inflammation both at the cellular and whole-body levels, and hence provide the microenvironment . AMPK is a critical regulator of glucose intake and energy balance and modulates energy regulation involved in glucose and lipid metabolism [18, 19]. On the other hand, AMPK has been identified as a novel target in tumor cell migration and invasion . Despite this, the role of AMPK activation in CCL3-mediated cancer migration has not been investigated in chondrosarcomas. In this study, we observed that CCL3 increases the migration of human chondrosarcoma cells and upregulates the expression of MMP-2. Furthermore, we observed that the CCR5 receptor, AMPK, p38, and NF-κB signaling pathways are involved in this process.
Materials and methods
Anti-mouse and anti-rabbit IgG-conjugated horseradish peroxidase, rabbit polyclonal antibodies specific for β-actin, AMPK, phospho(p)-AMPK (Thr172), p38, p-p38, inhibitor of kappa B (IκB), p-IκBα, IκB kinase (IKKα/β) (Ser180/181), NF-κB p65 subunit (p65), p-p65 (Ser536), and MMP-2; control shRNA (sc-108060) and CCL3 shRNA (sc-44722-SH) plasmids were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). AMPK inhibitors (Ara A and compound C), p38 inhibitor (SB203580), IκB protease inhibitor (TPCK), NF-κB inhibitor (PDTC), and MMP-2 inhibitor were purchased from Calbiochem (San Diego, CA). CCL3 and CCR5 monoclonal antibody (mAb) were purchased from Abcam (Cambridge, MA). Met-RANTES was purchased from R&D Systems (Minneapolis, MN, USA). The p38 dominant-negative MAPK mutant was provided by Dr. J. Han (Southwestern Medical Center, Dallas, TX). The IKKα (KM) (K44A) and IKKβ (KM) (K44A) mutants were gifted by Dr. H. Nakano (Juntendo University, Tokyo, Japan). All other chemicals were purchased from Sigma-Aldrich (St. Louis, MO).
The human chondrosarcoma cell line (JJ012) was kindly provided by the laboratory of Dr. Sean P. Scully (University of Miami School of Medicine, Miami, FL) and originated from Dr. Joel Block (Rush University Medical Center, Chicago, Illinois). JJ012 cells were cultured in a complete medium containing Dulbecco’s modified Eagle’s medium (DMEM)/α-minimum essential medium (α-MEM) supplemented with 10% fetal bovine serum (FBS). The human chondrosarcoma cell line (SW1353) was obtained from the American Type Culture Collection. SW1353 cells were cultured in a complete medium containing DMEM supplemented with 10% FBS. All experiments with cells were maintained at 37°C in a humidified atmosphere of 5% CO2.
CCL3 shRNA or control shRNA plasmids were transfected into JJ012 cells by using the transfection reagent, Lipofectamine 2000. At 24 h after transfection, stable transfectants were selected with 10 μg/mL puromycin (Life Technologies). The selection medium was replaced every 3 days and 2 weeks after selection, and puromycin-resistant cells and their clones were isolated.
Migration and invasion assay
The migration assay was performed using the Transwell assay (Costar, Acton, MA; pore size, 8-μm) in 24-well dishes. For the invasion assay, filters were precoated with 30 μL Matrigel Basement Membrane Matrix (BD Biosciences, Bedford, MA) for 30 min. The procedure for both migration and invasion assays was as follows. Before the migration assay was performed, cells were pretreated for 30 min with different concentrations of inhibitors, including the CCR5 mAb, compound C, Ara A, SB208530, TPCK, PDTC, MMP-2 inhibitor, or vehicle control (0.1% DMSO). Approximately 1 × 104 cells in 100 μL of serum-free medium was placed in the upper chamber, and 300 μL of the same medium containing CCL3 was placed in the lower chamber. The plates were incubated for 24 h at 37°C in 5% CO2, and cells were then fixed in methanol for 15 min and stained with 0.05% crystal violet in PBS for 15 min. Cells on the upper side of the filters were removed with cotton-tipped swabs, and the filters were washed with PBS. Cells on the underside of the filters were examined and counted under a microscope. Each clone was plated in triplicate in each experiment, and each experiment was repeated at least 3 times [3, 21].
Wound-healing migration assay
For wound-healing migration assays, cells were seeded on 12-well plates at a density of 1 × 105 cells/well in culture medium. At 24 h after seeding, the confluent monolayer of culture was scratched with a fine pipette tip, and migration was visualized by microscopy. The rate of wound closure was observed at the indicated times .
Supernatants collected from JJ012 cell cultures were mixed with sample buffer without reducing agents or heating. Samples were loaded onto a 10% SDS-PAGE gel containing 1 mg/ml gelatin and electrophoresed under constant voltage. Subsequently, the gel was washed with 2.5% Triton X-100 to remove SDS, rinsed with 50 mM Tris–HCl, pH 7.5, and then incubated overnight at room temperature with a developing buffer (50 mM Tris–HCl, pH 7.5, 5 mM CaCl2, 1 μM ZnCl2, 0.02% thimerosal, and 1% Triton X-100). Zymographic activity was revealed by staining with 1% Coomassie Blue.
Transfection of siRNAs or mutants
ON-TARGETplus siRNA targeting AMPKα1, AMPKα2 (The two catalytic subunit of AMPK; transfection of cells with AMPKα1 or AMPKα2 siRNA inhibited AMPKα1 or AMPKα2 expression, respectively; Additional file 1: Figure S1), MMP-2, and controls were purchased from Dharmacon Research (Lafayette, CO, USA). Transient transfection of siRNAs (10 nM) or dominant-negative mutants (0.5 μg) was carried out using DharmaFECT1 transfection reagent or Lipofectamine 2000 (Invitrogen, Carlsbad, CA), according to the manufacturer’s instructions, respectively.
Quantitative real-time polymerase chain reaction
Quantitative real-time polymerase chain reaction (qPCR) analysis was carried out using TaqMan® one-step PCR Master Mix (Applied Biosystems, Foster City, CA, USA). Total complementary DNA (100 ng/25 μL reaction) was mixed with sequence-specific primers and TaqMan® probes according to the manufacturer’s instructions. All target gene primers and probes were purchased commercially, and GAPDH was used as the internal control (Applied Biosystems). qPCR assays were carried out in triplicate with a StepOnePlus sequence detection system. The cycling conditions were 10 min polymerase activation at 95°C followed by 40 cycles at 95°C for 15 s and 60°C for 60 s. To calculate the cycle number at which the desired transcript was detected (denoted CT), the threshold was set above the non-template control background and within the linear phase of target gene amplification.
Western blot analysis
Cell lysates were prepared as described previously . Proteins were resolved by SDS-PAGE and transferred to polyvinyldifluoride (PVDF) membranes (Immobilon). The blots were blocked with 4% BSA for 1 h at room temperature and then probed with rabbit anti-human antibodies against MMP-2, p-IKKα/β, IKKα/β, p65, p-p65, p-AMPK, AMPK, p-p38, or p38 (1:1000) each separately for 1 h at room temperature. After three washes, the blots were incubated with peroxidase-conjugated donkey anti-rabbit secondary antibody (1:1000) for 1 h at room temperature. The blots were visualized by enhanced chemiluminescence using Kodak X-OMAT LS film (Eastman Kodak, Rochester, NY, USA).
Chromatin immunoprecipitation assay
Chromatin immunoprecipitation analysis was performed as described previously . DNA immunoprecipitated using the anti-p65 antibody was purified. The DNA was then extracted with phenol-chloroform. The purified DNA pellet was subjected to PCR. PCR products were resolved by 1.5% agarose gel electrophoresis and visualized by UV. The primers 5′-CCCCTGTTCAAGATGGAGTC-3′ and 5′-CCCAGGTTGCTTCCTTACCT-3′ were utilized in the PCR to amplify the human MMP-2 promoter region (−673 to −517) .
Human chondrosarcoma cells were transfected with a reporter plasmid using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s recommendations. At 24 h after transfection, the cells were pretreated with inhibitors for 30 min, and then, CCL3 or vehicle was added for 24 h. Cell extracts were then prepared, and luciferase and β-galactosidase activities were measured .
Human chondrosarcoma cells were plated on 24-well culture plates with coverslips. Cells were treated with CCL3 (30 ng/ml) and washed twice with ice-cold phosphate-buffered saline. Immunofluorescence staining using a primary anti-p65 monoclonal antibody was performed as described previously .
The human chondrosarcoma tissue array was purchased from Biomax (Rockville, MD, USA; 8 cases for normal cartilage, 4 cases for type Ib chondrosarcoma, and 4 cases for type IIb chondrosarcoma). The tissues were placed on glass slides, rehydrated, and incubated in 3% hydrogen peroxide to block the endogenous peroxidase activity. After trypsinization, sections were blocked by incubation in 3% bovine serum albumin (BSA) in PBS. The slides were incubated at 4°C overnight with a 1:50 diluted primary antibody, which was either monoclonal mouse anti-human CCL3, CCR5, or MMP-2 antibody. After being washed 3 times with PBS, samples were treated with goat anti-mouse IgG biotin-labeled secondary antibodies at a dilution of 1:50. The bound antibodies were detected with an ABC kit (Vector Laboratories, Burlingame, CA). The slides were stained with the chromogen diaminobenzidine, washed, counterstained with Delafield’s hematoxylin, dehydrated, treated with xylene, and mounted. The intensity of staining was evaluated as 0, 1+, 2+, 3+, 4+, and 5+ for no staining, very weak staining, weak staining, moderate staining, strong staining, and very strong respectively. IHC score was determined as the sum of the intensity score.
Data are presented as mean ± standard error of the mean (SEM). Statistical comparison of two groups was performed using the Student’s t test. Statistical comparisons of more than two groups were performed using one-way analysis of variance with Bonferroni’s post-hoc test. Analyzing patterns of staining in immunohistochemical studies statistical comparison of two tissue scores was performed using the Regression Analysis Method. In all cases, p < 0.05 was considered significant.
Correlation of CCL3, CCR5, and MMP-2 expression in human chondrosarcoma specimens
Involvement of MMP-2 in CCL3-induced migration of chondrosarcoma cells
Previous studies have shown significant expression levels of MMP-1, -2, -3, -9, and -13 in human chondrosarcoma cells [7, 27]. We therefore hypothesized that MMPs may be involved in CCL3-induced chondrosarcoma migration. Incubation of cells with CCL3 increased transcriptional expression of MMP-2 but not other MMPs, as measured by qPCR (Figure 2C). In JJ012 cells, CCL3 increased the expression of MMP-2 protein in a time-dependent manner (Figure 2D; upper panel). MMP-2 protein expression was also increased in the supernatant, and its enzyme activity was upregulated (Figure 2D; lower panel). To examine whether MMP-2 was involved in CCL3-induced cell migration, both the MMP-2 inhibitor and siRNA against MMP-2 were used. Pretreatment of cells with the MMP-2 inhibitor or transfection with MMP-2 specific siRNA abolished CCL3-induced cell migration and would healing activity (Figure 2E&F). To confirm that CCL3 mediates cell migration and MMP-2 expression in human chondrosarcoma cells, JJ012 cells lines stably expressing CCL3 shRNA were established. CCL3 expression in stable transfectants was compared with that in controls by western blotting. The expression of CCL3 was dramatically inhibited in JJ012/CCL3 shRNA cells (Figure 2I). While the knockdown of CCL3 did not affect JJ012 cell growth (data not shown), the migratory ability of JJ012 cells was inhibited (Figure 2G&H) as evaluated using Transwell assays and wound healing migration assays. In addition, CCL3 knockdown also reduced MMP-2 expression in JJ012 cells (Figure 2I). These results indicate that CCL3 upregulates MMP-2 and migration in chondrosarcoma cells, but not necessarily MMP-2 associated.
CCL3-mediated MMP-2 expression and cell migration are via the CCR5 receptor
The AMPK-dependent p38 pathway is involved in the CCL3-mediated cell migration and MMP-2 expression in chondrosarcoma cells
NF-κB signaling pathway is involved in CCL3-mediated MMP-2 up-regulation and migration activity
With the advent of systemic chemotherapy in the management of mesenchymal malignancies such as osteosarcoma and Ewing’s sarcoma, there has been a dramatic increase in the long-term survival of patients. In contrast, chondrosarcomas continue to have a poor prognosis owing to the absence of an effective adjuvant therapy . Chondrosarcoma shows a predilection for metastasis to the lungs and hence it is important to investigate the potential targets for preventing chondrosarcoma metastasis.
Immunohistochemical analyses revealed that the expression of CCL3 in chondrosarcoma patients was higher than that in normal cartilages. In this study, we hypothesized and investigated the paradigm that CCL3 may direct the metastasis of chondrosarcomas. Direct administration of exogenous CCL3 promoted cell migration, invasion, and wound healing activity in chondrosarcoma cells. On the other hand, CCL3-induced MMP-2 expression and cell motility were abolished by CCL3 shRNA. Our data suggest that CCL3 increases MMP-2 expression and subsequently promotes cell migration in human chondrosarcoma cells. The mechanisms underlying CCL3-induced increase in MMP-2 production and cell migration are activation of the CCR5 receptor, AMPK, p38, and NF-κB pathways.
As shown in previous studies, the CCR5 receptor is present on the surface of tumor cells and is responsible for CCL3-mediated cell motility . In this study, immunohistochemical staining revealed that high levels of CCL3 expression strongly correlated with CCR5 expression in human chondrosarcoma patients. Pre-treatment of chondrosarcoma cells with CCR5 mAb or inhibitor blocked CCL3-induced cell migration and reduced MMP-2 expression. Therefore, the CCL3-CCR5 interaction mediated the migratory activity in human chondrosarcoma cells.
Metastasis occurs in multiple steps. Tumor cells degrade the basement membrane, mainly through the use of MMPs . In human cancer cells, MMP-1, MMP-2, MMP-3, MMP-9, and MMP-13 have been found to be correlated with malignant grade and metastasis [4, 35]. On the other hand, activation of MMPs is also involved in CCL3-mediated cell motility . In this study, we found that while CCL3 induced MMP-2 expression and secretion in human chondrosarcoma cells, treatment of cells with the MMP-2 inhibitor reduced CCL3-induced cell migration. Moreover, inhibition of CCL3-enhanced MMP-2 protein expression, and siRNA knockdown of CCL-3 significantly suppressed CCL3-induced migration. Therefore, MMP-2 may be the CCL3-responsive mediator, and may degrade the ECM leading to subsequent cancer migration and metastasis.
AMPK is a heterotrimeric serine/threonine kinase composed of a catalytic subunit, α, and regulatory β and γ subunits . Previous studies have shown that AMPK is involved in the CCL3 signaling pathway . We observed that the AMPK inhibitors, namely, Ara A and compound C, antagonize CCL3-mediated cancer migration and MMP-2 expression, suggesting that AMPK activation is an obligatory event in CCL3-induced migration activity in these cells. We attempted to determine the identity of the catalytic subunit of AMPKα1 or AMPKα2, which mediates CCL3 signaling in human chondrosarcoma cells. We found that siRNA against both AMPKα1 and AMPKα2 reduce CCL3-mediated cancer migration, implying that AMPKα1 and AMPKα2 are involved in CCL3-induced migration activity. While adiponectin-mediated prostate cancer migration has been reported to be mediated by AMPKα1 but not AMPKα2 activation,  both AMPKα1 and AMPKα2 activation have been reported to mediate adiponectin-induced cell metastasis in human chondrosarcoma . These data indicate that AMPKα1 and AMPKα2 are important for metastasis of human chondrosarcoma. Since it has also been reported that AMPK interacts with p38 to regulate cell motility in human chondrosarcoma , we examined the potential role of p38 in the signaling pathway of CCL3-induced migration activity. Pre-treatment of chondrosarcoma cells for 30 min with SB203580 or transfection with the p38 mutant for 24 h markedly attenuated the CCL3-induced migration activity and MMP-2 expression. In addition, we observed that treatment of chondrosarcoma cells with CCL3 induced increased p38 phosphorylation. These effects were inhibited by Ara A or compound C. In contrast, the p38 inhibitor did not affect CCL3-promoted AMPK phosphorylation, indicating the involvement of AMPK-dependent p38 activation in CCL3-mediated migration and MMP-2 expression.
Tumor metastasis is the spread of tumor cells from a primary tumor to colonize other sites of the body. Invasion, intravasation, extravasation through the circulatory system, colonization, and finally angiogenesis at a distant site are the most common features of tumor metastasis. Because of the prevalence of distant metastasis in patients with chondrosarcoma, prognosis is generally very poor and the development of anti-metastatic therapy could be useful for these patients. Here, we found that CCL3 induced MMP-2 expression and subsequently promoted migration in human chondrosarcoma through activation of the CCR5 receptor, AMPK, p38, and NF-κB signaling pathways (Figure 7C). These observations may provide a better understanding of the mechanisms of metastasis and may lead to the development of effective therapies for chondrosarcoma.
This work was supported by grants from the National Science Council of Taiwan (NSC99-2320-B-039-003-MY3 and NSC100-2320-B-039-028-MY3) and China Medical University Hospital (DMR- 98–066). We thank Dr. H. Hakano for providing IKKα and IKKβ mutants; Dr. J. Han for providing p38 mutant.
- Terek RM, Schwartz GK, Devaney K, Glantz L, Mak S, Healey JH, Albino AP: Chemotherapy and P-glycoprotein expression in chondrosarcoma. J Orthop Res. 1998, 16: 585-590.PubMedView ArticleGoogle Scholar
- Fidler IJ: The organ microenvironment and cancer metastasis. Differentiation. 2002, 70: 498-505.PubMedView ArticleGoogle Scholar
- Wu MH, Lo JF, Kuo CH, Lin JA, Lin YM, Chen LM, Tsai FJ, Tsai CH, Huang CY, Tang CH: Endothelin-1 promotes MMP-13 production and migration in human chondrosarcoma cells through FAK/PI3K/Akt/mTOR pathways. J Cell Physiol. 2012, 227: 3016-3026.PubMedView ArticleGoogle Scholar
- Egeblad M, Werb Z: New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer. 2002, 2: 161-174.PubMedView ArticleGoogle Scholar
- Kerkela E, Saarialho-Kere U: Matrix metalloproteinases in tumor progression: focus on basal and squamous cell skin cancer. Exp Dermatol. 2003, 12: 109-125.PubMedView ArticleGoogle Scholar
- Chu CY, Cha ST, Chang CC, Hsiao CH, Tan CT, Lu YC, Jee SH, Kuo ML: Involvement of matrix metalloproteinase-13 in stromal-cell-derived factor 1 alpha-directed invasion of human basal cell carcinoma cells. Oncogene. 2007, 26: 2491-2501.PubMedView ArticleGoogle Scholar
- Hou CH, Hsiao YC, Fong YC, Tang CH: Bone morphogenetic protein-2 enhances the motility of chondrosarcoma cells via activation of matrix metalloproteinase-13. Bone. 2009, 44: 233-242.PubMedView ArticleGoogle Scholar
- Tsou HK, Chen HT, Hung YH, Chang CH, Li TM, Fong YC, Tang CH: HGF and c-Met interaction promotes migration in human chondrosarcoma cells. PLoS One. 2013, 8: e53974-PubMed CentralPubMedView ArticleGoogle Scholar
- Kulbe H, Levinson NR, Balkwill F, Wilson JL: The chemokine network in cancer–much more than directing cell movement. Int J Dev Biol. 2004, 48: 489-496.PubMedView ArticleGoogle Scholar
- Schall TJ, Bacon K, Camp RD, Kaspari JW, Goeddel DV: Human macrophage inflammatory protein alpha (MIP-1 alpha) and MIP-1 beta chemokines attract distinct populations of lymphocytes. J Exp Med. 1993, 177: 1821-1826.PubMedView ArticleGoogle Scholar
- Bennouna S, Bliss SK, Curiel TJ, Denkers EY: Cross-talk in the innate immune system: neutrophils instruct recruitment and activation of dendritic cells during microbial infection. J Immunol. 2003, 171: 6052-6058.PubMedView ArticleGoogle Scholar
- Nakasone Y, Fujimoto M, Matsushita T, Hamaguchi Y, Huu DL, Yanaba M, Sato S, Takehara K, Hasegawa M: Host-derived MCP-1 and MIP-1alpha regulate protective anti-tumor immunity to localized and metastatic B16 melanoma. Am J Pathol. 2012, 180: 365-374.PubMedView ArticleGoogle Scholar
- Arabzadeh A, Chan C, Nouvion AL, Breton V, Benlolo S, Demarte L, Turbide C, Brodt P, Ferri L, Beauchemin N: Host-related carcinoembryonic antigen cell adhesion molecule 1 promotes metastasis of colorectal cancer. Oncogene. 2013, 32: 849-860.PubMedView ArticleGoogle Scholar
- Wu Y, Li YY, Matsushima K, Baba T, Mukaida N: CCL3-CCR5 axis regulates intratumoral accumulation of leukocytes and fibroblasts and promotes angiogenesis in murine lung metastasis process. J Immunol. 2008, 181: 6384-6393.PubMedView ArticleGoogle Scholar
- Janciauskiene S, Wright HT: Inflammation, antichymotrypsin, and lipid metabolism: autogenic etiology of Alzheimer’s disease. Bioessays. 1998, 20: 1039-1046.PubMedView ArticleGoogle Scholar
- Yin K, Tang C: Inflammation, lipid metabolism dysfunction, and hypertension: active research fields in atherosclerosis-related cardiovascular disease in China. Sci China Life Sci. 2011, 54: 976-979.PubMedView ArticleGoogle Scholar
- Hursting SD, Digiovanni J, Dannenberg AJ, Azrad M, Leroith D, Demark-Wahnefried W, Kakarala M, Brodie A, Berger NA: Obesity, energy balance, and cancer: new opportunities for prevention. Cancer Prev Res (Phila). 2012, 5: 1260-1272.View ArticleGoogle Scholar
- Bijland S, Mancini SJ, Salt IP: Role of AMP-activated protein kinase in adipose tissue metabolism and inflammation. Clin Sci (Lond). 2013, 124: 491-507.View ArticleGoogle Scholar
- O’Neill HM: AMPK and exercise: glucose uptake and insulin sensitivity. Diabetes Metab J. 2013, 37: 1-21.PubMed CentralPubMedView ArticleGoogle Scholar
- Tang CH, Lu ME: Adiponectin increases motility of human prostate cancer cells via adipoR, p38, AMPK, and NF-kappaB pathways. Prostate. 2009, 69: 1781-1789.PubMedView ArticleGoogle Scholar
- Tan TW, Yang WH, Lin YT, Hsu SF, Li TM, Kao ST, Chen WC, Fong YC, Tang CH: Cyr61 increases migration and MMP-13 expression via alphavbeta3 integrin, FAK, ERK and AP-1-dependent pathway in human chondrosarcoma cells. Carcinogenesis. 2009, 30: 258-268.PubMedView ArticleGoogle Scholar
- Wu MH, Chen LM, Hsu HH, Lin JA, Lin YM, Tsai FJ, Tsai CH, Huang CY, Tang CH: Endothelin-1 enhances cell migration through COX-2 up-regulation in human chondrosarcoma. Biochim Biophys Acta. 2013, 1830: 3355-3364.PubMedView ArticleGoogle Scholar
- Tang CH, Hsu CJ, Fong YC: The CCL5/CCR5 axis promotes interleukin-6 production in human synovial fibroblasts. Arthritis Rheum. 2010, 62: 3615-3624.PubMedView ArticleGoogle Scholar
- Huang CY, Chen SY, Tsai HC, Hsu HC, Tang CH: Thrombin induces epidermal growth factor receptor transactivation and CCL2 expression in human osteoblasts. Arthritis Rheum. 2012, 64: 3344-3354.PubMedView ArticleGoogle Scholar
- Hou CH, Chiang YC, Fong YC, Tang CH: WISP-1 increases MMP-2 expression and cell motility in human chondrosarcoma cells. Biochem pharmacol. 2011, 81: 1286-1295.PubMedView ArticleGoogle Scholar
- Roberti MP, Arriaga JM, Bianchini M, Quinta HR, Bravo AI, Levy EM, Mordoh J, Barrio MM: Protein expression changes during human triple negative breast cancer cell line progression to lymph node metastasis in a xenografted model in nude mice. Cancer Biol Ther. 2012, 13: 1123-1140.PubMed CentralPubMedView ArticleGoogle Scholar
- Tan TW, Lai CH, Huang CY, Yang WH, Chen HT, Hsu HC, Fong YC, Tang CH: CTGF enhances migration and MMP-13 up-regulation via alphavbeta3 integrin, FAK, ERK, and NF-kappaB-dependent pathway in human chondrosarcoma cells. J Cell Biochem. 2009, 107: 345-356.PubMedView ArticleGoogle Scholar
- Zucchetto A, Benedetti D, Tripodo C, Bomben R, Dal Bo M, Marconi D, Bossi F, Lorenzon D, Degan M, Rossi FM, et al: CD38/CD31, the CCL3 and CCL4 chemokines, and CD49d/vascular cell adhesion molecule-1 are interchained by sequential events sustaining chronic lymphocytic leukemia cell survival. Cancer research. 2009, 69: 4001-4009.PubMedView ArticleGoogle Scholar
- Kim Y: Regulation of cell proliferation and migration in glioblastoma: new therapeutic approach. Front Oncol. 2013, 3: 53-PubMed CentralPubMedGoogle Scholar
- Chiu YC, Shieh DC, Tong KM, Chen CP, Huang KC, Chen PC, Fong YC, Hsu HC, Tang CH: Involvement of AdipoR receptor in adiponectin-induced motility and alpha2beta1 integrin upregulation in human chondrosarcoma cells. Carcinogenesis. 2009, 30: 1651-1659.PubMedView ArticleGoogle Scholar
- Omoike OI, Teague RM, Benedict SH, Chan MA: MIP-1alpha induces binding of nuclear factors to the kappaB DNA element in human B cells. MCBRC. 2000, 4: 15-19.PubMedGoogle Scholar
- Fong YC, Yang WH, Hsu SF, Hsu HC, Tseng KF, Hsu CJ, Lee CY, Scully SP: 2-methoxyestradiol induces apoptosis and cell cycle arrest in human chondrosarcoma cells. J Orthop Res. 2007, 25: 1106-1114.PubMedView ArticleGoogle Scholar
- Menu E, De Leenheer E, De Raeve H, Coulton L, Imanishi T, Miyashita K, Van Valckenborgh E, Van Riet I, Van Camp B, Horuk R, et al: Role of CCR1 and CCR5 in homing and growth of multiple myeloma and in the development of osteolytic lesions: a study in the 5TMM model. Clin Exp Metastasis. 2006, 23: 291-300.PubMedView ArticleGoogle Scholar
- Deryugina EI, Quigley JP: Matrix metalloproteinases and tumor metastasis. Cancer Metastasis Rev. 2006, 25: 9-34.PubMedView ArticleGoogle Scholar
- Scherer RL, McIntyre JO, Matrisian LM: Imaging matrix metalloproteinases in cancer. Cancer Metastasis Rev. 2008, 27: 679-690.PubMedView ArticleGoogle Scholar
- Repeke CE, Ferreira SB, Claudino M, Silveira EM, de Assis GF, Avila-Campos MJ, Silva JS, Garlet GP: Evidences of the cooperative role of the chemokines CCL3, CCL4 and CCL5 and its receptors CCR1+ and CCR5+ in RANKL + cell migration throughout experimental periodontitis in mice. Bone. 2010, 46: 1122-1130.PubMedView ArticleGoogle Scholar
- Gruzman A, Babai G, Sasson S: Adenosine monophosphate-activated protein kinase (AMPK) as a new target for antidiabetic drugs: a review on metabolic, pharmacological and chemical considerations. Rev Diabet Stud. 2009, 6: 13-36.PubMed CentralPubMedView ArticleGoogle Scholar
- Taddei SR, Queiroz CM, Moura AP, Andrade I, Garlet GP, Proudfoot AE, Teixeira MM, da Silva TA: The effect of CCL3 and CCR1 in bone remodeling induced by mechanical loading during orthodontic tooth movement in mice. Bone. 2013, 52: 259-267.PubMedView ArticleGoogle Scholar
- Chen YJ, Wei YY, Chen HT, Fong YC, Hsu CJ, Tsai CH, Hsu HC, Liu SH, Tang CH: Osteopontin increases migration and MMP-9 up-regulation via alphavbeta3 integrin, FAK, ERK, and NF-kappaB-dependent pathway in human chondrosarcoma cells. J Cell Physiol. 2009, 221: 98-108.PubMedView ArticleGoogle Scholar
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