MMP-9 Gene Expression Research Articles

The combined treatment of brassinin and imatinib synergistically downregulated the expression of MMP-9 in SW480 colon cancer cells.

In cancer treatment, which is a major cause of mortality today, combination studies with clinically used chemotherapeutics are becoming increasingly important as much as investigating the effects of novel natural compounds. In this context, phytoalexins constitute an important group due to their unique structure. Brassinin is an essential indole phytoalexin and is a biosynthetic precursor for other phytoalexins. The purpose of this study was to evaluate the anticancer effects of brassinin in combination with imatinib in SW480 cells. In the study, it was observed that brassinin-imatinib combination significantly increased cytotoxicity compared with the single treatment of both compounds and inhibited cell cycle at G0/G1 phase. Annexin V binding and fluorescence imaging assays showed that the combination of brasinin-imatinib induces apoptosis in a dose-dependent manner. Furthermore, the effect of brassinin on the activity of MMP-9 in SW480 cells was evaluated for the first time, and it was detected that MMP-9 activity was significantly reduced. The combination of brassinin-imatinib was found to inhibit MMP-9 activity as well as relative MMP-9 gene expression on a higher level compared with control and compounds alone. Our findings have revealed that the combination of brassinin-imatinib synergistically induces cytotoxicity and apoptosis in SW480 cells. The findings on MMP-9 downregulation have also revealed the anti-metastatic potential of treatment.

Inhibition of the Expression of Matrix Metalloproteinases in Articular Chondrocytes by Resveratrol through Affecting Nuclear Factor-Kappa B Signaling Pathway.

In the present study, we tried to examine whether resveratrol regulates the expression of matrix metalloproteinases (MMPs) through affecting nuclear factor-kappa B (NF-κB) in articular chondrocytes. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-1β (IL-1β)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. Effect of resveratrol on IL-1β-induced secretion of MMP-3 was investigated in rabbit articular chondrocytes using western blot analysis. To elucidate the action mechanism of resveratrol, effect of resveratrol on IL-1β-induced NF-κB signaling pathway was investigated in SW1353, a human chondrosarcoma cell line, by western blot analysis. The results were as follows: (1) resveratrol inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) resveratrol reduced the secretion of MMP-3; (3) resveratrol inhibited IL-1β-induced activation (phosphorylation) of inhibitory kappa B kinase (IKK), and thus phosphorylation and degradation of inhibitory kappa Bα (IκBα); (4) resveratrol inhibited IL-1β-induced phosphorylation and nuclear translocation of NF-κB p65. This, in turn, led to the down-regulation of gene expression of MMPs in SW1353 cells. These results suggest that resveratrol can regulate the expression of MMPs through affecting NF-κB by directly acting on articular chondrocytes.

Maldevelopment of the submandibular gland in a mouse model of apert syndrome.

Apert syndrome is characterized by craniosynostosis and bony syndactyly of the hands and feet. The cause of Apert syndrome is a single nucleotide substitution mutation (S252W or P253R) in fibroblast growth factor receptor 2 (FGFR2). Clinical experience suggests increased production of saliva by Apert syndrome patients, but this has not been formally investigated. FGFR2 signaling is known to regulate branching morphogenesis of the submandibular glands (SMGs). With the Apert syndrome mouse model (Ap mouse), we investigated the role of FGFR2 in SMGs and analyzed the SMG pathology of Apert syndrome. Ap mice demonstrated significantly greater SMG and sublingual gland (SMG/SLG complex) mass/body weight and percentage of parenchyma per unit area of the SMG compared with control mice. Furthermore, gene expression of Fgf1, Fgf2, Fgf3, Pdgfra, Pdgfrb, Mmp2, Bmp4, Lama5, Etv5, and Dusp6 was significantly higher in the SMG/SLG complex of Ap mice. FGF3 and BMP4 exhibited altered detection patterns. The numbers of macrophages were significantly greater in SMGs of Ap mice than in controls. Regarding functional evaluations of the salivary glands, no significant differences were observed. These results suggest that the gain-of-function mutation in FGFR2 in the SMGs of Ap mice enhances branching morphogenesis. Developmental Dynamics 247:1175-1185, 2018. © 2018 Wiley Periodicals, Inc.

Ski drives an acute increase in MMP-9 gene expression andrelease in primary cardiac myofibroblasts.

Many etiologies of heart disease are characterized by expansion and remodeling of the cardiac extracellular matrix (ECM or matrix) which results in cardiac fibrosis. Cardiac fibrosis is mediated in cardiac fibroblasts by TGF‐β 1/R‐Smad2/3 signaling. Matrix component proteins are synthesized by activated resident cardiac fibroblasts known as myofibroblasts (MFB). These events are causal to heart failure with diastolic dysfunction and reduced cardiac filling. We have shown that exogenous Ski, a TGF‐β 1/Smad repressor, localizes in the cellular nucleus and deactivates cardiac myofibroblasts. This deactivation is associated with reduction of myofibroblast marker protein expression in vitro, including alpha smooth muscle actin (α‐SMA) and extracellular domain‐A (ED‐A) fibronectin. We hypothesize that Ski also acutely regulates MMP expression in cardiac MFB. While acute Ski overexpression in cardiac MFB in vitro was not associated with any change in intracellular MMP‐9 protein expression versus LacZ‐treated controls,exogenous Ski caused elevated MMP‐9 mRNA expression and increased MMP‐9 protein secretion versus controls. Zymographic analysis revealed increased MMP‐9‐specific gelatinase activity in myofibroblasts overexpressing Ski versus controls. Moreover, Ski expression was attended by reduced paxillin and focal adhesion kinase phosphorylation (FAK ‐ Tyr 397) versus controls. As myofibroblasts are hypersecretory and less motile relative to fibroblasts, Ski's reduction of paxillin and FAK expression may reflect the relative deactivation of myofibroblasts. Thus, in addition to its known antifibrotic effects, Ski overexpression elevates expression and extracellular secretion/release of MMP‐9 and thus may facilitate internal cytoskeletal remodeling as well as extracellular ECM components. Further, as acute TGF‐β 1 treatment of primary cardiac MFB is known to cause rapid translocation of Ski to the nucleus, our data support an autoregulatory role for Ski in mediating cardiac ECM accumulation.

Short-term Effects of Whey, Creatine, and L-carnitine Supplementation on Muscle Hypertrophy Marker Candidates in Young Males: A Randomized Placebo-controlled Pilot Study

Previous research showed that resistance exercise could induce muscle development, noticeably from the increased level of several markers from blood samples. However, no study had been performed to explore the effect of combination of resistance exercise and proper nutrition supply on those markers. The aim of this study to investigate the effect of whey protein, creatine, and L-carnitine; on potential molecular markers of muscle hypertrophy (arg1 and mmp9) from blood samples. Twelve healthy male participants were randomly categorized into supplement (SUPP) or placebo (PLAC) treatment, and performed resistance training three times in a one-week period. Blood sampling was carried out before (day one) and 2 hours after the exercise (day one, day three and day five). The level of mmp9 gene expression was increased along with the progress of the resistance training program. Moreover, participants who received supplementation (SUPP) showed a higher level of mmp9 gene expression compared to resistance training only (PLAC). A significant difference was observed between two treatments in the first day, 2 hours after the resistance training session (p = .04); and between SUPP group on the fifth day, 2 hours after the resistance training; compared to the first day, before the resistance training session (p = .02). The effect was not observed on arg1 gene. A combination of resistance training with supplementation; was considered to enhance the muscle hypertrophy process, compared to resistance training only. The results also suggested that mmp9 could act as a blood-derived molecular marker of muscle hypertrophy.

Methylsulfonylmethane and mobilee prevent negative effect of IL-1β in human chondrocyte cultures via NF-κB signaling pathway

Nutraceuticals are compounds that serve as nutrition with an easy accessibility and favourable safety profile. Recent studies showed their potential activity on osteoarthritis (OA) inflammation and cartilage metabolism.We investigated the effect of methylsulfonylmethane (MSM) and mobilee in human OA chondrocyte cultures exposed to interleukin (IL)-1β.OA cartilage was obtained from femoral heads of five patients undergoing total replacement surgery. Chondrocytes were incubated with mobilee (200 and 500 μM) and MSM (2000 and 6000 μM) in presence of IL-1β (10 ng/mL) and nuclear factor (NF)-κB inhibitor (BAY 11-7082, 1 μM), for 24 and 48 h. Viability and apoptosis were performed by MMT and flow cytometry. The metalloproteinase (MMP)-1,-3,-13 and type II collagen (Col2a1) were analyzed by qRT-PCR and ELISA, and NF-κB activation by immunofluorescence.IL-1β stimulus determined a significant regulation of survival, apoptotic ratio, as well as of gene expression and serum levels of MMP-1,-3,-13 and Col2a1 in OA chondrocytes compared to baseline. Mobilee and MSM incubation significantly reversed the effect of IL-1β. IL-1β significantly induced NF-κB p50 nuclear translocation, which was significantly counteracted by the pre-treatment of OA chodrocytes with the tested compounds. BAY11-7082 significantly modulated MMPs and Col2a1 expression respectively to basal state. Co-treatment of IL-1β with mobilee, MSM and BAY11-7082 didn't cause changes of MMPs or Col2a1 beyond that caused by each single treatment.We demonstrated that MSM and mobilee have a beneficial effect on OA chondrocytes metabolism, probably due to the modulation of NF-κB pathway, providing a powerful rationale for the use of these substances in OA treatment.

Surface Nanocoating with Plant‐Derived Pectins Improves Fibroblast Response In Vitro

Pectins, plant‐derived polysaccharides, are novel candidates for biomaterial nanocoatings. Their chemical structure imitates the polysaccharides from the extracellular matrix of mammals, providing bio‐specific cell adhesion. It is possible to control the structure of pectin rhamnogalacturonan‐I (RG‐I) by enzymatic modification. RG‐Is has been proposed for surface nanocoating of orthopedic and dental titanium implants due to its effect on osseointegration with stimulation of bone and connective tissue healing. The aim of the current study is to evaluate in vitro, the impact of plant‐derived pectin RG‐I nanocoating on the cellular response of human fibroblasts. In this study, unmodified RG‐I and enzymatically modified RG‐I with low amount of arabinose and high amount of galactose are examined. All in vitro studies are performed on tissue culture polystyrene plates, whose surfaces are coated with unmodified and modified RG‐I. The results show that the nanocoating with pectin RG‐Is increas proliferation, cell metabolic activity, and gene expression of COL1A1, FN1, FGFR1, BMP7, and MMP2 as compared to control surface, without pectin coatings. The results show that nanocoating with RG‐Is has the capacity to improve fibroblast proliferation, differentiation, and extracellular matrix protein production, and may therefore be considered as a potential candidate for further improvement of bone and connective tissue regeneration.

Activation of NF-κB in Synovium versus Cartilage from Patients with Advanced Knee Osteoarthritis: A Potential Contributor to Inflammatory Aspects of Disease Progression.

The aim was to assess the activation and association of the NF-κB system across synovial membrane (SM) and articular cartilage (AC) in patients with knee osteoarthritis (OA) and ascertain its potential effects on catabolic mediator expression in advanced OA. SM and AC were obtained from 40 OA patients undergoing total knee arthroplasty and from 19 postmortem control subjects. NF-κB subunit RelA in nuclear and cytosolic fractions and NF-κB1-DNA binding in nuclear extracts was assessed by ELISA, whereas NFKB1, RELA, IL-8, IL-6, and MMP3 gene expression were analyzed by reverse transcriptase-quantitative PCR in tissues. We observed higher SM nuclear RelA protein levels and upregulated NF-κB1-DNA binding in OA patients compared with postmortem controls. However, in AC, lower nuclear RelA levels were observed compared with cytosolic extracts in patients. Nuclear RelA levels correlated positively with NF-κB1-DNA binding in SM and AC in patients. SM RELA and MMP3 mRNA levels were upregulated, whereas IL-8 and IL-6 as well as AC RELA were downregulated in patients compared with controls. In SM, nuclear RelA levels correlated positively with MMP3 gene expression in patients. A negative correlation was observed between SM nuclear RelA levels and AC NF-κB1-DNA binding, and SM nuclear NF-κB1-DNA binding correlated negatively with AC MMP3 and NFKB1 mRNA levels in patients. These findings highlight NF-κB-triggered cross-talk and feedback mechanisms between SM and AC in OA. Further, our findings strongly support a role for an activated NF-κB system in the transcriptional mechanism of inflammatory processes, especially in SM of patients with advanced OA.

Involvement of Catechols in Acteoside in the Activation of Promatrix Metalloproteinase-2 and Membrane Type-1-Matrix Metalloproteinase Expression via a Phosphatidylinositol-3-Kinase Pathway in Human Dermal Fibroblasts.

Granulation tissue formation during skin wound healing requires the migration and proliferation of dermal fibroblasts in the wound site, where a subsequent remodeling of extracellular matrices (ECM) occurs. An abnormality of ECM remodeling within the healing wound leads to fibrosis and a contracted scar. To evaluate whether acteoside, a phenylethanoid glycoside isolated from the leaves of Rehmannia glutinosa LIBOSCH., exhibits wound-healing actions, we examined the effect of acteoside on the expression of matrix metalloproteinases (MMPs) in normal human dermal fibroblasts (NHDF). Acteoside dose- and time-dependently augmented the activation of the precursor of MMP-2 (proMMP-2/progelatinase A) in untreated- and interleukin-1β-treated NHDF, while the alteration of the MMP-2 gene expression was negligible. The acteoside-induced proMMP-2 activation was associated with the augmented membrane-type 1 MMP (MT1-MMP) expression in the NHDF. In addition, the proMMP-2 activation was enhanced by two aglycones in acteoside: caffeic acid and 3,4-dihydroxyphenylethanol, which consist of catechol. However, there was no change in the proMMP-2 activation in other catechol derivatives: homovanillyl alcohol- and homovanillic acid-treated NHDF, indicating that catechols in acteoside were requisite for the regulation of the MMP activation and expression in NHDF. Furthermore, the proMMP-2 activation by acteoside was selectively inhibited by LY294002, a potent phosphatidylinositol-3-kinase (PI3K) inhibitor. These results provide novel evidence that acteoside augments proMMP-2 activation along with an increase in MT1-MMP expression through a PI3K signal pathway in NHDF. Thus, acteoside is likely to be an attractive candidate that facilitates ECM remodeling in the skin wound repair process.

Tc17/IL-17A Up-Regulated the Expression of MMP-9 via NF-κB Pathway in Nasal Epithelial Cells of Patients With Chronic Rhinosinusitis.

Chronic rhinosinusitis (CRS) is a common chronic inflammatory disease of the upper airways involving nasal cavity and sinus. Deriving both from its clinical complexity with protean clinical manifestations as well its pathogenetic heterogeneity, the molecular mechanisms contributing to the pathogenesis of CRS remain unclear, and attract a wide interest in the field. Current evidences indicate that IL-17A is highly expressed in chronic rhinosinusitis with nasal polyps (CRSwNP). However, its pathogenetic role in regulation of tissue remodeling of CRSwNP remains unknown. The present study aimed to investigate the cellular origins and functions of IL-17A cytokine in CRSwNP, and further determined whether IL-17A could affect the expression of metalloproteinases (MMPs), the remodeling factors of CRSwNP. The results showed that the expression of IL-17A was upregulated in nasal tissues of patients with CRSwNP compared to those with chronic rhinosinusitis without nasal polyps (CRSsNP) and controls. CD8+ cytotoxic T lymphocytes (Tc) were major IL-17A producers in nasal tissues of CRSwNP. Interleukin (IL)-17-producing CD8+ T cells (Tc17) was significantly higher in nasal tissues of CRSwNP than CRSsNP and controls. Nonetheless, no difference was observed among the IL-17A in peripheral blood lymphocytes of these three groups. Moreover, in the same patients, IL-17A expression was negligible in lymphocytes of peripheral blood when compared with nasal tissues. Increased gene and protein expression of MMP-7 and MMP-9 in patients with CRSwNP compared with controls were observed. In CRSwNP samples, IL-17A receptor (IL-17AR) co-localized with MMP-9 and they were mainly expressed in the epithelial cells. MMP-9 expression was up-regulated both in Primary human nasal epithelial cells (PHNECs) and a nasal epithelial cell line (RPMI 2650) by IL-17A treatment, and diminished by anti-IL-17AR treatment. Furthermore, IL-17A promoted the expression of MMP-9 by activating the NF-κB signal pathway. Thus, our results have revealed a crucial role of IL-17A and Tc cells on pathogenesis and tissue remodeling of CRSwNP.

Gender-specific effects of MMP-2 and MMP-9 gene variants and the risk of aneurysmal subarachnoid haemorrhage

Matrix metalloproteinases (MMPs), a family of zinc-dependent proteases, have been linked with the pathogenesis of intracranial aneurysm (IA) formation and rupture. Patients with IA have elevated serum levels of MMP-2 and MMP-9 gene expression has been shown to be increased in aneurysm tissue. In this study, we have evaluated the association between MMP gene variants and aneurysmal subarachnoid hemorrhage (aSAH). The rs243865 in MMP-2 and rs17576 in MMP-9 genes were genotyped using Taqman allelic discrimination assay. Upon gender stratification, the presence of G allele of MMP-9 gene exon variant was found to increase the risk of aSAH by 1.4 fold (p = 0.032), in men. The GG genotype also showed a trend towards association with the risk of aSAH (OR: 1.785, CI: 0.971–3.283, p = 0.062) in men, after adjusting for the vascular risk factors. In women, no association was observed. The MMP-2 gene variants were not associated with aSAH in both genders. Further functional studies are required to describe the exact basis of the association between MMP-9 polymorphism and intracranial aneurysm in men.

Dimetallic Ru(II) arene complexes appended on bis-salicylaldimine induce cancer cell death and suppress invasion via p53-dependent signaling

A series of bis-salicylaldimine ligands bearing two ON-donor functions were reacted with dichloro(p-cymene)ruthenium(II) dimer in the presence of base (NaOAc) and a series of four dimetallic Ru(II) arene complexes (Ru(p-cymene))2(bis-salicylaldimine)Cl2 (C1C4) were prepared. These complexes were obtained in excellent isolated yields and characterized in detail by using different spectroscopic techniques. The structure of C1 was also determined in solid state by single crystal X-ray analysis. These complexes were studied for their cytotoxic effect against three different types of human cancer cells including hepatocellular carcinoma (HepG2), non-small-cell lung cancer (A549) and breast cancer (MCF-7) cells by MTT assay. These complexes showed considerable cytotoxic effect in all the above-mentioned cell lines that was comparable to the effect of cisplatin. C1 and C2 showed moderate anticancer effect while C3 and C4 showed reasonable cytotoxicity. We found the cytotoxicity was increased in series from C1 to C4 representing the effect of ligand modification from small to bulky group at the amine functionality of the salicylaldimine. We selected C3 and C4 for mechanistic anticancer study in MCF-7 cells. The acridine orange/ethidium bromide and DAPI staining assays of MCF-7 cells treated with Ru(II) complexes showed apoptosis in cancer cells. Similarly, these complexes induced p53 protein expression in MCF-7 cells. Further, increased mRNA levels of p63, p73, PUMA, BAX and NOXA genes were observed in response to the treatment with C3 and C4, while cyclinD1, MMP3 and ID1 gene expression was significantly reduced. We found reduced invasion ability in breast cancer cells treated with C3 and C4. Taken together, we demonstrated that bis-salicylaldimine based dimetallic Ru-(p-cymene) complexes exerts anticancer effects by p53 pathway, suggesting the promising chemotherapeutic potentials of these Ru(II) complexes for the treatment of cancer. This study may further pave for their in depth in vitro or in vivo anticancer investigations.

Platelet-Derived Growth Factor Receptor-α and β are Involved in Fluid Shear Stress Regulated Cell Migration in Human Periodontal Ligament Cells.

Fluid shear stress (FSS) is the most common stress produced by mastication, speech, or tooth movement. However, how FSS regulates human periodontal ligament (PDL) cell proliferation and migration as well as the underlying mechanism remains unknown. FSS (6dyn/cm2) was produced in a flow chamber. Cell proliferation was tested by the 5-ethynyl-2'-deoxyuridine assay. Cell migration was tested by the wound healing assay. Gene and protein expression of platelet-derived growth factors (PDGFs) and matrix metalloproteinase (MMP)-2 were measured by reverse transcription-polymerase chain reaction and western blot analyses. We investigated the effect of 4h of 6dyn/cm2 FSS on proliferation and migration of PDL cells. FSS promoted PDL cell proliferation but inhibited migration. The gene and protein expression of PDGF receptor (PDGFR)-α and β both decreased in response to FSS. Activating and inhibiting the PDGFRs did not affect the FSS-induced increase in cell proliferation. However, activating PDGFRs with PDGF-BB, which bound both PDGFR-α and β, and PDGF-CC and DD, which had high affinities for PDGFR-α and PDGFR-β, individually rescued FSS-inhibited migration. FSS also inhibited MMP-2 gene expression, which was the most important factor for matrix turnover and migration of PDLs. PDGF-BB, CC, and DD increased the FSS-induced decline in MMP-2 expression. These results indicate that MMP-2 is regulated by FSS and contributes to the FSS-induced decrease in cell migration. Our study suggests a role for PDGFR-α and β in short-term FSS-regulated cell proliferation and migration. These results will help provide the scientific foundation for revealing the mechanisms clinical tooth movement and PDL regeneration.

MicroRNAs and Osteoarthritis.

An imbalance in gene expressional events skewing chondrocyte anabolic and catabolic pathways toward the latter causes an aberrant turnover and loss of extracellular matrix proteins in osteoarthritic (OA) articular cartilage. Thus, catabolism results in the elevated loss of extracellular matrix proteins. There is also evidence of an increase in the frequency of chondrocyte apoptosis that compromises the capacity of articular cartilage to undergo repair. Although much of the fundamental OA studies over the past 20 years identified and characterized many genes relevant to pro-inflammatory cytokines, apoptosis, and matrix metalloproteinases (MMPs)/a disintegrin and metalloproteinase with thrombospondin motif (ADAMTS), more recent studies focused on epigenetic mechanisms and the associated role of microRNAs (miRs) in regulating gene expression in OA cartilage. Thus, several miRs were identified as regulators of chondrocyte signaling pathways, apoptosis, and proteinase gene expression. For example, the reduced expression of miR-146a was found to be coupled to reduced type II collagen (COL2) in OA cartilage, whereas MMP-13 levels were increased, suggesting an association between MMP-13 gene expression and COL2A1 gene expression. Results of these studies imply that microRNAs could become useful in the search for diagnostic biomarkers, as well as providing novel therapeutic targets for intervention in OA.

The effects of mid- and long-term endurance exercise on heart angiogenesis and oxidative stress.

Objective(s):Long-term, irregular endurance exercise may result in disturbance to the angiogenesis of heart muscles and blood supply. The aim of the present study is to evaluate the effects of mid- and long-term endurance exercise on the process of angiogenesis.Materials and Methods:Eighteen male Wister rats of 220±10 g, were randomly assigned to three groups of 6 rats including: Control, Mid, and Long Group. After the training sessions, the rats were weighed and sacrificed.Results:In comparison to the Control Group, the both groups, indicated remarkable increase in the weight of heart and significantly higher serum LDH and CK activity (P<0.01). In addition, after the training sessions, weakened antioxidant heart system (TAC, total thiol groups, and GPX activity) and increased oxidative stress markers (MDA and NO) were remarkably observed in Mid Group and particularly in those in the Long Group in comparison to the Control Group (P<0.05). Finally, significant increase in VEGF-B, MEF-2C and MMP-2 gene expression was found for both experimental groups, associated with the up-regulation of ANGPT-1 and HDAC4 in the Mid Group (P<0.05). While the longer exercise period induced significantly upper VEGF-B, MEF-2C, and MMP-2 and significantly lower ANGPT-1 and HDAC4 in the Long Group (P<0.05). Conclusion:In this study, higher oxidative status and upper angiogenic gene expression with higher VEGF-B, MEF-2c, and MMP-2 and lower ANGPT-1 and HDAC4 were traced as effects of long-term endurance exercise. These results point to the dis-regulation of blood supply in the presence of angiogenesis resulting from long-term exercise.

Expression and correlation of MMP-9, VEGF, and p16 in infantile hemangioma.

The incidence and progression of infantile hemangioma depend on several factors. The objective of the present study was to analyze the expression of MMP-9, VEGF, and p16 in hemangioma, and evaluate the clinical significance of the correlation between the gene and protein expression of MMP-9 and VEGF. 60 pathological tissue samples from patients with hemangioma (30 cases of proliferative hemangioma and 30 cases of degenerative hemangioma) and 30 normal epidermal tissue samples were collected. Tissues were examined by immunohistochemistry. Additionally, MMP-9 and VEGF gene expression were evaluated by qRT-PCR to determine the correlation between gene and protein level. Immunohistochemistry showed that the expression of MMP-9 and VEGF was higher in proliferative hemangioma, while the expression of p16 was lower, and the differences were statistically significant (p<0.05). According to qRT-PCR, MMP-9 and VEGF gene expression were positively correlated with their protein expression (p<0.05). The tumor suppressor gene p16 is negatively correlated with the occurrence and degradation of hemangioma. Furthermore, the gene and protein expression of MMP-9 and VEGF in infantile hemangioma are positively correlated. The levels of MMP-9 and VEGF are higher in the proliferative stage of hemangioma than other stages. Both MMP-9 and VEGF facilitate angiogenesis and tumorigenesis, and are likely to serve as markers for the development of hemangioma in the future.

Modulation of matrix metabolism by ATP-citrate lyase in articular chondrocytes

Certain dysregulated chondrocyte metabolic adaptive responses such as decreased activity of the master regulator of energy metabolism AMP-activated protein kinase (AMPK) promote osteoarthritis (OA). Metabolism intersects with epigenetic and transcriptional responses. Hence, we studied chondrocyte ATP-citrate lyase (ACLY), which generates acetyl-CoA from mitochondrial-derived citrate, and modulates acetylation of histones and transcription factors. We assessed ACLY in normal and OA human knee chondrocytes and cartilages by Western blotting and immunohistochemistry, and quantified acetyl-CoA fluorometrically. We examined histone and transcription factor lysine acetylation by Western blotting, and assessed histone H3K9 and H3K27 occupancy of iNOS, MMP3, and MMP13 promoters by chromatin immunoprecipitation (ChIP) and quantitative PCR (qPCR). We analyzed iNOS, MMP3, MMP13, aggrecan (ACAN), and Col2a1 gene expression by RT-qPCR. Glucose availability regulated ACLY expression and function, nucleocytosolic acetyl-CoA, and histone acetylation. Human knee OA chondrocytes exhibited increased ACLY activation (assessed by Ser-455 phosphorylation), associated with increased H3K9 and H3K27 acetylation. Inhibition of ACLY attenuated IL-1β-induced transcription of iNOS, MMP3, and MMP13 by suppressing acetylation of p65 NF-κB, H3K9, and H3K27, blunted release of NO, MMP3, and MMP13, and also reduced SOX9 acetylation that promoted SOX9 nuclear translocation, leading to increased aggrecan and Col2a1 mRNA expression. ACLY is a novel player involved in regulation of cartilage matrix metabolism. Increased ACLY activity in OA chondrocytes increased nucleocytosolic acetyl-CoA, leading to increased matrix catabolism via dysregulated histone and transcription factor acetylation. Pharmacologic ACLY inhibition in OA chondrocytes globally reverses these changes and stimulates matrix gene expression and AMPK activation, supporting translational investigation in OA.

Fisetin inhibits cell migration via inducing HO-1 and reducing MMPs expression in breast cancer cell lines

Metastasis is commonly seen in advanced stage of cancers, and matrix metalloproteinases (MMPs) are commonly up-regulated and have been identified as critical regulators. In this present study, a flavonoid, fisetin, which can be found in diverse foods, is investigated for its ability to inhibit cell motility, and the underlying mechanism is also studied in breast cancer cells (4T1 and JC cells). We have revealed that fisetin increased HO-1 mRNA and protein expressions. Besides, fisetin also elevated Nrf2 expression in nuclear fraction. By silencing Nrf2, fisetin-induced HO-1 expression was abrogated, suggested that HO-1 expression was mediated by up-regulation of the transcription factor Nrf2. In addition, we also found that fisetin decreased MMP-2 and MMP-9 enzyme activity and gene expression in both protein and mRNA levels. Moreover, by administration of HO-1 inhibitors, tin protoporphyrin and zinc protoporphyrin, fisetin-reduced MMP-2 and MMP-9 expressions were reversed. Furthermore, transfection of siRNA against HO-1 and Nrf2 also abolished MMP-2 and MMP-9 reduction exerted by fisetin. These findings suggest that fisetin-mediated MMP-2 and MMP-9 reduction is regulated by HO-1 through Nrf2. Therefore, fisetin may be useful as a potential therapeutic agent for the treatment of metastatic breast cancer.

Static magnetic field regulates proliferation, migration, differentiation and YAP/TAZ activation of human dental pulp stem cells

The dental pulp stem cells (DPSCs) are a population of mesenchymal stem cells, which have multilineage potential and high proliferation. DPSCs are regarded as a promising tool for tissue regeneration of dentine, dental pulp, bone, cartilage, and muscle. Recently, magnetic materials have become commonly applied in dental clinics. Static magnetic field has been reported to regulate the proliferation, migration, or differentiation of stem cells. However, whether static magnetic fields affect DPSCs is still unknown. In our study, we investigated the effect of static magnetic field on the proliferation, migration, and differentiation of DPSCs. The results indicated that static magnetic field rearranged the cytoskeleton of DPSCs. A static magnetic field of 1mT increased DPSC proliferation, as well as the gene expression of several growth factors such as FGF-2, TGF-β, and VEGF. Moreover, the static magnetic field promoted the migration of DPSCs by regulating MMP-1 and MMP-2 gene expression. Static magnetic field of 1mT also induced osteo/odontogenesis and mineralization in DPSCs. Otherwise, the static magnetic field recruited YAP/TAZ to the nucleus, inhibited the phosphorylation of YAP/TAZ, and upregulated the two YAP/TAZ-regulated genes, CTGF and ANKRD1. Cytoskeleton inhibitor, cytochalasin D, obviously inhibited the nuclear localization of YAP/TAZ. When YAP/TAZ were knocked-down, the static magnetic field-induced mineralization of DPSCs was diminished. Our findings provide an insight into the effect of static magnetic field on DPSCs and provide the foundation for the future tissue regeneration.

Upregulating MMP-1 in carcinoma-associated fibroblasts reduces the efficacy of Taxotere on breast cancer synergized by Collagen IV.

Chemotherapy is an important comprehensive treatment for breast cancer, which targets micro-environment of tumors as well as their characterisitcs. A previous microarray analysis revealed that matrix metalloproteinase (MMP)-1 was highly upregulated in carcinoma-associated fibroblasts (CAFs) prior to and following treatment with Taxotere under co-culture conditions. However, whether the chemotherapeutic effects of Taxotere were influenced by the changes in MMP-1 remained unclear. The purpose of the present study was to investigate the impact and mechanism of CAFs in regulating the efficacy of Taxotere on breast cancer cells. CAFs isolated from primary invasive ductal human breast tumors following surgical resection, were used in co-culture with MDA-MB-231 cells to simulate the tumor micro-environment. Following the addition of Taxotere, changes in MMP-1 gene and protein expression were assessed by reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. Proliferation, invasion and apoptosis assays revealed that when MMP-1 was upregulated in CAFs, the therapeutic efficacy of Taxotere was reduced in breast cancer cells. Chemosensitivity was significantly increased when MMP-1 expression was inhibited by GM6001. In addition, Collagen IV was upregulated in CAFs following chemotherapy and protected breast cancer cells against chemotherapeutic side effects. Collagen IV expression significantly decreased, as well as MMP-1 expression when GM6001 was added. Proliferation and invasion assays demonstrated that the exogenous addition of Collagen IV weakend the chemotherapeutic effect of Taxotere on breast tumor cells. Overall, the results revealed that in CAFs, MMP-1 synergized with Collagen IV as a key gene in regulating the chemotherapeutic effect of Taxotere on breast tumor cells and served an important role in reducing the efficacy of Taxotere on breast cancer, potentially via the transforming growth factor-β signaling pathway. These fidings provide a theoretical basis for the mechanism of CAFs in reducing the chemotherapeutic effect of Taxotere on breast cancer cells and a novel approach for enhancing the chemosensitivity of tumors.