MMP-2 Activity Research Articles

Silibinin Inhibits Cell Invasion through the Inhibition of MMPs, p-p38, and IL-1β in Human Fibrosarcoma Cells

Normal cells become tumorigenic owing to mutations in oncogenes and tumor suppressor genes modulating cell division. Cancer cells break down extracellular matrix to metastasize other tissues. Therefore, the development of natural and synthetic substances that suppress metastatic enzymes such as matrix metalloproteinase (MMP)-2 and MMP-9 is useful to inhibit metastasis. Silibinin is the main ingredient of silymarin extracted from the seeds of milk thistle plants having lung cancer-suppressing effects and liver protection. The purpose of this study was to investigate the inhibitory effect of silibinin on the invasion of human fibrosarcoma cells. The effect of silibinin on cell viability was measured in HT1080 cells using an MTT assay. The MMP-9 and MMP-2 activities were analyzed using a zymography assay. The expression of proteins in cytoplasm related to metastasis was examined by western blot analysis and immunofluorescence assay. In this study, silibinin above 20 μM showed growth inhibitory effects. Silibinin above 20 μM remarkably inhibited the levels of MMP-2 and MMP-9 activation under phorbol myristate acetate (PMA) treatment conditions. Furthermore, silibinin at 25 μM reduced the levels of MMP-2, IL-1β, ERK-1/2, and p-p38 expression and silibinin above 10 μM inhibited cell invasion on HT1080 cells. These findings indicate that silibinin may have an inhibitory effect on the enzymes involved in invasion, hence it might influence the metastatic ability of tumor cells.

Abstract 2757: Combination of a fluorescent substrate-based MMP activity assay and hit pick reading/imaging procedure to screen for inhibitors of MT1-MMP activity and 3D glioma tumoroid invasion

Abstract Malignant and nonmalignant brain tumors are composed of a group of more than 100 different types. Malignant gliomas, including glioblastoma multiforme (GBM) and astrocytomas, are the most common primary brain tumors in the United States, and make up 78% of malignant brain tumors. Membrane type 1 matrix metalloproteinase (MT1-MMP) has been shown to be crucial for the progression, invasion, migration, and angiogenesis of tumors. Use of a FRET substrate to quantify MT1-MMP activity, where tumoroids are embedded in a collagen hydrogel, would simplify detection of potential protease inhibitors, while using an in vivo-like cell model. In the present study, multiple glioma cell lines were formed into tumor spheroids, followed by type I collagen addition. Many known MT1-MMP inhibitors were then added to the wells, to potentially inhibit invasion of the 3D cell models. The substrate and experimental setup were then combined with the joint reading and imaging process available in the Agilent BioTek Gen5 microplate reader and imager software, to create a hit pick screening method. During the procedure, all control and test wells were read, and the signal from cleaved substrate was quantified. Uninhibited invasion within positive control wells exhibited high fluorescence values. Therefore, a hit pick criterion was established such that any wells containing test molecules that were statistically lower than average in fluorescent signal from the corresponding positive control, which should exhibit inhibited invasion, were imaged. Wells that were not significantly lower were not imaged. The result was a screening method which increased efficiency by reducing image capture time and storage requirements. Citation Format: Brad Larson, Gregg Fields, Ania Knapinska. Combination of a fluorescent substrate-based MMP activity assay and hit pick reading/imaging procedure to screen for inhibitors of MT1-MMP activity and 3D glioma tumoroid invasion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2757.

Abstract 5823: Uncovering the mechanisms of matrix metalloproteinase-3 (MMP-3) in the cisplatin resistance of ovarian cancer

Abstract Seventy percent of women with ovarian cancer will develop chemotherapy-resistance to cisplatin. Regardless of the advancements in technology, mortality rates for ovarian cancer have declined only slightly in the last forty years, reiterating the need to generate novel treatment options targeting cisplatin resistance. In RNA-seq and microarrays experiments we observed that Matrix-metalloproteinase-3 (MMP3) was highly abundant in cisplatin-resistant when compared to cisplatin-sensitive cells. MMP3 is a member of the matrix metalloproteinases (MMPs) family of proteolytic enzymes, involved in the degradation of proteins in the extracellular matrix (ECM). Deregulation of MMPs alters the extracellular matrix and contributes to proliferation, invasion, and metastasis of cancerous cells. A typical MMP structure consists of a pro-domain, a catalytic, and a hemopexin domain. The catalytic domain is responsible for MMPs proteolytic activity, while the hemopexin domain contributes to substrate and ligand specificity. Inhibitors targeting the catalytic domain of MMPs were unsuccessful in clinical trials due to a lack of selectivity and specificity. When we used a chemical inhibitor, we did not observe reduced cell-proliferation or invasiveness. However, MMP3-targeted small interference RNA (siRNA) was able to reduce cell proliferation and invasive capabilities in cisplatin resistant ovarian cancer cells. Additionally, MMP3-targeted siRNA significantly reduced the sensitivity of ovarian cancer cells to cisplatin treatment. Cisplatin also increased MMP3 mRNA levels in ovarian cancer mice models and human cell lines. This evidence suggests that the MMP3 levels correlate with the sensitivity of ovarian cancer cells to cisplatin treatment. We hypothesize that the hemopexin domain will bind to proteins promoting cisplatin-resistance. We validated our expression profile studies at mRNA and protein levels via western blot, ELISA and qRT-PCR. Additionally, survival analysis using the Kaplan-Meier plotter database showed that ovarian cancer patients with low MMP3 levels live longer than patients with higher MMP3 levels. Furthermore, when we measured MMP3 activity, we observed decreased activity in cisplatin-resistant versus cisplatin-sensitive cells. Taken together, these data suggest that other domains of MMP3 (i.e., hemopexin domain) may have a function in the cisplatin resistance of ovarian cancer cells. Citation Format: Mariela Rivera-Serrano, Pablo E. Vivas-Mejia. Uncovering the mechanisms of matrix metalloproteinase-3 (MMP-3) in the cisplatin resistance of ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5823.

Involvement of matrix metalloproteinase 1 and urokinase-type plasminogen activator in the PKCα-p38 MAPK pathway-mediated progression of human liver cancer cells.

Our previous studies have shown that the plasminogen activator (PA) and matrix metalloproteinases (MMPs) proteinase systems were highly expressed in highly malignant liver cancer cells and regulated by PKCα. This study investigates whether the PKCα regulation of PA and MMPs systems is conducted through p38 mitogen-activated protein kinase (MAPK) signaling and the pathway is responsible for promoting cell progression. We found that the expressions of p38 MAPK in both highly malignant HA22T/VGH and SK-Hep-1 liver cancer cells were higher than that in other lower malignancy liver cancer cells. Since PKCα activates p38 MAPK in progression of liver cancer, we suspected the PKCα/p38 MAPK signaling pathway to be involved in the regulation of MMPs and PA systems. When SK-Hep-1 cells were treated with SB203580 or DN-p38, only MMP-1 and u-PA mRNA expressions decreased. The p38 MAPK inhibition also decreased the cell migration and invasion. In addition, the mRNA decay assays showed that the higher expressions of MMP-1 and u-PA mRNA in SK-Hep-1 cells were due to the alteration of mRNA stability by p38 MAPK inhibition. Zymography of SK-Hep-1 cells treated with siPKCα vector also showed the decrease of the activity of MMP-1 and u-PA and confirmed changes in mRNA level. Furthermore, only the transfection of MKK6 to the siPKCα-treated SK-Hep-1 stable clone cell restored the attenuation of MMP-1 and u-PA expressions. The treatment of SK-Hep-1 cells with either inhibitor of MMP-1 or u-PA reduced migration, and the reduction was enhanced with both inhibitors. In addition, tumorigenesis was also reduced with both inhibitors. These data suggest a novel finding that MMP-1 and u-PA are critical components in PKCα/MKK6/p38 MAPK signaling pathway which mediates liver cancer cell progression, and that the targeting of both genes may be a viable approach in liver cancer treatment.

Comparison of DPPIV Levels in Serum and Tumour of OSCC Patients and Its Correlation with Active Matrix Metalloproteinases 2 and 9.

The important role of Dipeptidyl Peptidase IV (DPPIV) has been reported in tumour progression of several human cancers. This study demonstrates the DPPIV mRNA expression level and activity in tumour and paired non-tumour tissues of oral squamous cell carcinoma (OSCC) patients and the potential modulation of DPPIV in the metastasis of tumour through regulating MMP2 and MMP9 activities. This study was conducted on 16 OSCC patients. The mRNA expression level of DPPIV was evaluated by RT-qPCR in tumour of OSCC patientsand compared with their paired non-tumour tissues. Additionally, DPPIV activity was measured in serum, tumour and paired non-tumour tissues of OSCC patients. Zymography was performed to measure and compare the activities of MMP2 and MMP9 between tumour and paired non-tumour tissues of OSCC patients. The results showed significantly higher DPPIV mRNA level and activity in tumour of OSCC patients compared to their paired non-tumour tissues. Tumour DPPIV mRNA expression and activity were positively correlated with activities of MMP2 and MMP9, respectively. Serum DPPIV activity of OSCC patients was lower compared to healthy control and did not show correlation with tumour DPPIV mRNA level. These data indicate that secreted DPPIV may not originate from the tumour tissue of OSCC patients. Furthermore, increased DPPIV gene expression and activity in tumour of OSCC patients might be involved in the ECM degradation and invasion of OSCC through regulation of MMP2 and MMP9 activities.

Retraction notice to Glucose transporter-1 gene expression is associated with pancreatic cancer invasiveness and MMP-2 activity [Surgery -September 2004, Pages 548-556

This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the Editors-in-Chief, because an investigation by Harvard Medical School and the Brigham and Women’s Hospital has concluded that Figure 3 is invalid and no underlying research data are available to resolve the discrepancies or validate the reported results. Glucose transporter-1 gene expression is associated with pancreatic cancer invasiveness and MMP-2 activitySurgeryVol. 136Issue 3PreviewOverexpression of the facilitative glucose transporter-1 (GLUT-1) has been observed for a wide range of human cancers, with the degree of overexpression generally being inversely correlated with prognosis. We tested the effects of modulating GLUT-1 expression on pancreatic cancer cellular invasiveness. Full-Text PDF

Screening of MMP-2 Inhibiting Phytoconstituents for the Development of Newer Pancreatic Cancer Treatment Modalities.

Pancreatic cancer metastasis is characterized by a higher incidence of morbidity and mortality. The present study attempts to identify phytocomponents with the potential to inhibit the secretion of MMP-2 by pancreatic cancer cells and ascertain the efficacy of individual components. Overall survival analysis carried out revealed reduced survival of patients with high MMP-2 expression. Data analysis from TCGA revealed increased MMP-2 expression in pancreatic cancer patients compared to adjacent normal tissues. The expression of MMP-2 was reported at different stages of pancreatic cancer (Stage I-IV). To understand the relevance of phytocomponents in binding to the catalytic site of MMP-2, molecular docking studies were performed to find the effectiveness based on Glide score/energy. To substantiate the in-silico analysis, the eight components were also tested in vitro for reducing the survival in PANC-1 cells at three different time points (24, 48, and 72 hours). Finally, zymography analysis was performed using the eight components in the PANC-1 conditioned media of treated cells to ascertain the enzymatic activity of MMP-2. The obtained results suggest plumbagin, emodin, and EGCG exert potential inhibition in PANC-1 cells, among other phytocomponents tested. Therefore, as assessed using computational studies, the binding ability of plumbagin, emodin, and EGCG can be interpreted as inhibiting effects on MMP-2 activities. These compounds could find potential application in preventing the progression, sustenance, and metastasis of pancreatic cancer and need to be explored further using a pre-clinical model system in order to validate the efficacy, bioavailability, and safety.

Vernodalin Suppresses Tumor Proliferation and Increases Apoptosis of Gastric Cancer Cells Through Attenuation of FAK/PI3K/AKT/mTOR and MAPKs Signaling Pathways.

Gastric cancer (GC) is the most aggressive malignant tumor with limited treatment alternatives post metastasis. Vernodalin (VN) induced apoptosis has been reported in various types of human cancer cells. However, the precise molecular mechanisms underlying the anti-metastasis action of VN on GC cells are yet to be elucidated. In this study, we investigated the anti-metastatic and apoptotic effects of VN on SGC- 7901 and AGS cells, with a purpose of gaining a deeper understanding of the anti-metastatic mechanisms of VN on gastric carcinoma. To attenuate the activation of PI3K/AKT/mTOR and mitogen-activated protein kinase (MAPK) signaling pathways by VN in GC cells. We employed VN and gastric cancer cells in experiments such as MTT assay, apoptosis, MMP, DAPI, Rh-123, cell adhesion assay, and western blot analysis on GC SGC-7901 and AGS cells. Our results revealed that VN inhibits cell proliferation, adhesion, and metastasis and induces apoptosis of both GC cells. VN potentially reduced the protein expressions of MMP-2, MMP-9, and uPA, whereas intensified expressions of TIMP-1 and TIMP-2. Also, VN attenuates the expression of FAK, p-PI3K, p-AKT, p-mTOR, p-JNK, p-p38MAPK, and p-ERK. Thus, it is inferred that VN treatment reduced the activities of MMP-2 and MMP-9 via the FAK/PI3K/AKT/ mTOR, and MAPKs signaling pathways. Our results confirm that VN prevented GC growth, invasion and metastasis and induce apoptosis in GC cells. Our findings suggest that VN is a potential natural therapeutic compound as a new remedy for GC chemotherapy treatment.

Protease-Triggered, Spatially Controlled DNA Assembly in Apoptotic Cells for Early Evaluation of Therapeutic Efficacy.

Despite numerous advances in the use of DNA as building blocks to assemble complex structures, the dearth of strategies that allow for protease-controlled in situ DNA assembly in living cells remains a bottleneck in this field. Here, we present a modular engineering approach to achieve protease-triggered self-assembly of DNA in apoptotic cells for early evaluation of tumor response to drug treatment. In the design, peptide nucleic acid is introduced as a building bridge to engineer DNA building blocks with peptides and thus to suppress their self-assembly activity, while caspase-3 (Casp-3) protease-mediated enzymatic cleavage of the peptide substrate enables the activation of the DNA assembly, generating fluorescence signal output for real-time monitoring of Casp-3 activity. Furthermore, the specific protease triggering imparts DNA assembly with spatial selectivity to apoptotic cells in vivo, allowing for early evaluation of tumor therapeutic efficacy. Moreover, the strategy is extended to probe the activity of MMP-2 for lymph node metastasis imaging, demonstrating the universality of this approach. This work highlights protease-controlled DNA assembly in ways that are simple and versatile, with the potential to expand the repertoire of DNA nanotechnology for diverse biomedical applications.

A new approach for the synthesis of tri-substituted pyrazole propionic acids derivatives: Anti-inflammatory, antimicrobial and molecular docking studies

The present study involves design, synthesis and biological evaluation of novel 3-(3,5-diphenyl-1H-pyrazol-4-yl)-3-(aryl)propanoic acid 6(a-f) and propanoate 7(a-f) derivatives which are analogous to the known drug lonazolac. The pyrazole derivative of lonazolac exhibiting interesting biological activities, such as antimicrobial, antifungal and anti-inflammatory etc. Therefore, the targeted moieties were synthesised in their pure form without the use of column chromatography. All the synthesised moieties were examined against MMP-2 and MMP-9 anti-inflammatory activity in comparison with tetracycline. Compound 6(a-f) series showed more than 80% and 70% inhibition against MMP-2 and MMP-9 respectively. Wherein, 7(a-f) series showed moderate percentage of inhibition against both MMP-2 and MMP-9. The antimicrobial activity was performed by micro-broth dilution assay using Itraconazole and ciprofloxacin as reference. The targets have shown promising biological activities. Docking studies of the molecule on crystal structure of E. coli have shown good docking score with similar interactions as that of the standard clorobiocin.

A self-etch bonding system with potential to eliminate selective etching and resist proteolytic degradation

ObjectivesBonded restorations using self-etch (SE) systems exhibit a limited lifespan due to their susceptibility to hydrolytic, enzymatic or fatigue degradation and poor performance on enamel. This study was conducted to develop and assess the performance of a two-step SE system using a functional monomer bis[2-(methacryloyloxy)ethyl]phosphate (BMEP) and demonstrate a strategy to enhance stability of bonded resin composite restorations to both enamel and dentine. MethodsA two-step SE system was formulated with a primer containing BMEP, with an adhesive with or without BMEP, and compared to a commercial 10-MDP-containing system, ClearfilTM SE Bond 2 (CFSE). The systems were evaluated on enamel for surface roughness and microshear bond strength (µSBS) and on dentine for microtensile bond strength (µTBS), nanoleakage, MMP inhibition and cyclic flexural fatigue. ResultsWhilst all bonding systems resulted in statistically similar µSBS, BMEP-based primers yielded greater enamel surface roughness than the CFSE primer. The BMEP-free adhesives resulted in statistically similar or higher µTBS and lower nanoleakage compared to CFSE. In situ zymography revealed minimal to no MMP activity within the hybrid layer of BMEP-based systems. The BMEP-free adhesive exhibited flexural strength and fatigue resistance statistically similar to CFSE. ConclusionsIncorporation of BMEP in the primer led to satisfactory bond strengths with both enamel and dentine, potentially eliminating the need for selective enamel etching. Combined with an adhesive formulation that is solvent-free and hydrophobic, and confining the acidic functional monomer in the primer resulted in minimal interfacial leakage, and resistance to proteolytic degradation and the cyclic nature of chewing. Clinical significanceThe SE bonding system containing BMEP combines the potent etching of phosphoric acid with the therapeutic function of the phosphate-based monomer in creating a homogenous hybrid layer with protection against endogenous proteolytic enzymes. This strategy may overcome current challenges that arise during selective enamel etching.

Chemical profile, in vitro pharmacological activity and Satureja cuneifolia Ten. evaluation of essential oil based on distillation time

ABSTRACT The medicinal plant Satureja cuneifolia Ten. was widely utilized as spice, tea and traditional medicine. The objective of the current study was to examine the chemical composition and in vitro biological activities (LOX, MMP-1, and MMP-12 enzyme inhibition activity and cytotoxicity on A549 cell line) of Satureja cuneifolia extracts and essential oils. The essential oils of the flowering aerial parts were hydro-distilled at four different distillation times (5, 30, 60, and 180 min) using the Clevenger apparatus. The total essential oil and four fragments were compared in terms of the major component, yield, and distillation time. Volatile compounds of the infusion were extracted by using HS-SPME. Ethanolic extract had the strongest inhibition activity on the LOX enzyme (84.50%), while the essential oils exhibited more cytotoxic activity on the A549 cell line than the extracts. The oils and the infusion were analyzed using GC-MS and the primary chemicals identified by LC-MS/MS.

The protective effects of a novel AT2 receptor agonist, β-Pro7Ang III in ischemia-reperfusion kidney injury

Background and purposeThis study investigated the reno-protective effects of a highly selective AT2R agonist peptide, β-Pro7Ang III in a mouse model of acute kidney injury (AKI). MethodsC57BL/6 J mice underwent either sham surgery or unilateral kidney ischemia-reperfusion injury (IRI) for 40 min. IRI mice were treated with either β-Pro7Ang III or perindopril and at 7 days post-surgery the kidneys analysed for histopathology and the development of fibrosis and matrix metalloproteinase (MMP)-2 and -9 activity. The association of the therapeutic effects of β-Pro7Ang III with macrophage number and phenotype was determined in vivo and in vitro. Key resultsDecreased kidney tubular injury, interstitial matrix expansion and reduced interstitial immune cell infiltration in IRI mice receiving β-Pro7Ang III treatment was observed at day 7, compared to IRI mice without treatment. This correlated to reduced collagen accumulation and MMP-2 activity in IRI mice following β-Pro7Ang III treatment. FACS analysis showed a reduced number and proportion of CD45+CD11b+F4/80+ macrophages in IRI kidneys in response to β-Pro7Ang III, correlating with a significant increase in M2 macrophage markers and decreased M1 markers at day 3 and 7 post-IR injury, respectively. In vitro analysis of cultured THP-1 cells showed that β-Pro7Ang III attenuated lipopolysaccharide (LPS)-induced tumour necrosis factor-α (TNF-α) and interleukin (IL)− 6 production but increased IL-10 secretion, compared to LPS alone. ConclusionAdministration of β-Pro7Ang III via mini-pump improved kidney structure and reduced interstitial collagen accumulation, in parallel with an alteration of macrophage phenotype and anti-inflammatory cytokine release, therefore mitigating the downstream progression of ischemic AKI.

Exposure to oxLDL impairs TGF-β activity in human tendon cells

BackgroundPrevious studies have shown that patients with hypercholesterolemia experience elevated levels of oxidized LDL (oxLDL), a molecule which triggers inflammation and collagenase activity. In this study we discovered novel mechanistic effects of oxLDL on tendon cells and the mediators regulating matrix remodeling by analyzing the expression and activity of related proteins and enzymes. These effects may contribute to tendon damage in patients with high cholesterol.MethodsIsolated human tendon cells (male and female donors age 28 ± 1.4 age 37 ± 5.7, respectively) were incubated in the presence or absence of oxLDL. The influence of oxLDL on the expression level of key mRNA and proteins was examined using real time quantitative PCR, ELISA and Western blots. The activities of enzymes relevant to collagen synthesis and breakdown (lysyl oxidase and matrix metalloproteinases) were quantified using fluorometry. Finally, the isolated human tendon cells in a 3D construct were exposed to combinations of oxLDL and TGF-β to examine their interacting effects on collagen matrix remodeling.ResultsThe one-way ANOVA of gene expression indicates that key mRNAs including TGFB, COL1A1, DCN, and LOX were significantly reduced in human tendon cells by oxLDL while MMPs were increased. The oxLDL reduced the activity of LOX at 50 µg/ml, whereas conversely MMP activities were induced at 25 µg/ml (P ≤ 0.01). COL1A1 synthesis and TGF-β secretion were also inhibited (P ≤ 0.05). Adding recombinant TGF-β reversed the effects of oxLDL on the expression of collagens and LOX. OxLDL also impaired collagen matrix remodeling (P ≤ 0.01), and adding TGF-β restored the native phenotype.ConclusionExposure to oxLDL in patients with hypercholesterolemia may adversely affect the mechanical and structural properties of tendon tissue through a direct action of oxLDL on tendon cells, including impairment of TGF-β expression. This impairment leads to disturbed matrix remodeling and synthesis, thereby potentially leading to increased risk of acute or chronic tendon injury. Our discovery may provide an opportunity for developing effective treatments for tendon injury in hypercholesterolemia patients by targeting the TGF-β pathway.

Platelet-derived microparticles stimulate the invasiveness of colorectal cancer cells via the p38MAPK-MMP-2/MMP-9 axis

BackgroundMetastasis is the main cause of death in patients with colorectal cancer (CRC). Apart from platelets, platelet-derived microparticles (PMPs) are also considered important factors that can modify the activity of cancer cells. PMPs are incorporated by cancer cells and can also serve as intracellular signalling vesicles. PMPs are believed to affect cancer cells by upregulating their invasiveness. To date, there is no evidence that such a mechanism occurs in colorectal cancer. It has been shown that platelets can stimulate metalloproteases (MMPs) expression and activity via the p38MAPK pathway in CRC cells, leading to their elevated migratory potential. This study aimed to investigate the impact of PMPs on the invasive potential of CRC cells of various phenotypes via the MMP-2, MMP-9 and p38MAPK axis.MethodsWe used various CRC cell lines, including the epithelial-like HT29 and the mesenchymal-like SW480 and SW620. Confocal imaging was applied to study PMP incorporation into CRC cells. The presence of surface receptors on CRC cells after PMP uptake was evaluated by flow cytometry. Transwell and scratch wound-healing assays were used to evaluate cell migration. The level of C-X-C chemokine receptor type 4 (CXCR4), MMP-2, and MMP-9 and the phosphorylation of ERK1/2 and p38MAPK were measured by western blot. MMP activity was determined using gelatine-degradation assays, while MMP release was evaluated by ELISA.ResultsWe found that CRC cells could incorporate PMPs in a time-dependent manner. Moreover, PMPs could transfer platelet-specific integrins and stimulate the expression of integrins already present on tested cell lines. While mesenchymal-like cells expressed less CXCR4 than epithelial-like CRC cells, PMP uptake did not increase its intensity. No significant changes in CXCR4 level either on the surface or inside CRC cells were noticed. Levels of cellular and released MMP-2 and MMP-9 were elevated in all tested CRC cell lines after PMP uptake. PMPs increased the phosphorylation of p38MAPK but not that of ERK1/2. Inhibition of p38MAPK phosphorylation reduced the PMP-induced elevated level and release of MMP-2 and MMP-9 as well as MMP-dependent cell migration in all cell lines.ConclusionsWe conclude that PMPs can fuse into both epithelial-like and mesenchymal-like CRC cells and increase their invasive potential by inducing the expression and release of MMP-2 and MMP-9 via the p38MAPK pathway, whereas CXCR4-related cell motility or the ERK1/2 pathway appears to not be affected by PMPs.A1m_Mu21TUHDLKeRdJvkXhVideo

Protease-Activatable Porphyrin Molecular Beacon for Osteoarthritis Management.

Despite the substantial burden posed by osteoarthritis (OA) globally, difficult challenges remain in achieving early OA diagnosis and adopting effective disease-modifying treatments. In this study, we use a biomolecular approach to address these limitations by creating an inherently theranostic molecular beacon whose imaging and therapeutic capabilities are activated by early pathological changes in OA. This platform comprised (1) a peptide linker substrate for metalloproteinase-13 (MMP-13), a pathological protease upregulated in OA, which was conjugated to (2) a porphyrin moiety with inherent multimodal imaging, photodynamic therapy, and drug delivery capabilities, and (3) a quencher that silences the porphyrin's endogenous fluorescence and photoreactivity when the beacon is intact. In diseased OA tissue with upregulated MMP-13 expression, this porphyrin molecular beacon (PPMMP13B) was expected to undergo sequence-specific cleavage, yielding porphyrin fragments with restored fluorescence and photoreactivity that could, respectively, be used as a readout of MMP-13 activity within the joint for early OA imaging and disease-targeted photodynamic therapy. This study focused on the synthesis and characterization of PPMMP13B, followed by a proof-of-concept evaluation of its OA imaging and drug delivery potential. In solution, PPMMP13B demonstrated 90% photoactivity quenching in its intact form and robust MMP-13 activation, yielding a 13-fold increase in fluorescence post-cleavage. In vitro, PPMMP13B was readily uptaken and activated in an MMP-13 cell expression-dependent manner in primary OA synoviocytes without exuding significant cytotoxicity. This translated into effective intra-articular cartilage (to a 50 μm depth) and synovial uptake and activation of PPMMP13B in a destabilization of the medial meniscus OA mouse model, yielding strong fluorescence contrast (7-fold higher signal than background) at the diseased joint site. These results provide the foundation for further exploration of porphyrin molecular beacons for image-guided OA disease stratification, effective articular delivery of disease-modify agents, and OA photodynamic therapy.

The differential role of resistin on invasive liver cancer cells.

To determine whether inhibition of kinase signaling will suppress resistin-induced liver cancer progression. Resistin is located in monocytes and macrophages of adipose tissue. This adipocytokine is an important link between obesity, inflammation, insulin resistance, and cancer risk. Pathways that resistin is known to be involved include but are not limited to mitogen-activated protein kinases (MAPKs) and extracellular signal-regulated kinases (ERK). The ERK pathway promotes cellular proliferation, migration, survival of cancer cells, and tumor progression. The Akt pathway is known to be up-regulated in many cancers including liver cancer. Using an invitro model, HepG2 and SNU-449 liver cancer cells were exposed to resistin±ERK, Akt, or both inhibitors. The following physiological parameters were assessed: cellular proliferation, ROS, lipogenesis, invasion, MMP, and lactate dehydrogenase activity. The inhibition of kinase signaling suppressed resistin-induced invasion and lactate dehydrogenase in both cell lines. In addition, in SNU-449 cells, resistin increased proliferation, ROS, and MMP-9 activity. Inhibition of PI3K and ERK decreased phosphorylated Akt and ERK, and pyruvate dehydrogenase. In this study, we describe the effect of Akt andERK inhibitors to determine if inhibition suppresses resistin-induced liver cancer progression. Resistin promotes cellular proliferation, ROS, MMP, invasion and LDH activity in SNU-449 liver cancer cells which is differentially mediated by Akt and ERK signaling pathways.

Citral Modulates MMP-2 and MMP-9 Activities on Healing of Gastric Ulcers Associated with High-Fat Diet-Induced Obesity.

Obesity causes low-grade inflammation that results in the development of comorbidities. In people with obesity, exacerbation of gastric lesion severity and delayed healing may aggravate gastric mucosal lesions. Accordingly, we aimed to evaluate the citral effects on gastric lesion healing in eutrophic and obese animals. C57Bl/6 male mice were divided into two groups: animals fed a standard diet (SD) or high-fat diet (HFD) for 12 weeks. Gastric ulcers were induced using acetic acid (80%) in both groups. Citral (25, 100, or 300 mg/kg) was administered orally for 3 or 10 days. A vehicle-treated negative control (1% Tween 80, 10 mL/kg) and lansoprazole-treated (30 mg/kg) were also established. Lesions were macroscopically examined by quantifying regenerated tissue and ulcer areas. Matrix metalloproteinases (MMP-2 and -9) were analyzed by zymography. The ulcer base area between the two examined periods was significantly reduced in HFD 100 and 300 mg/kg citral-treated animals. In the 100 mg/kg citral-treated group, healing progression was accompanied by reduced MMP-9 activity. Accordingly, HFD could alter MMP-9 activity, delaying the initial healing phase. Although macroscopic changes were undetectable, 10-day treatment with 100 mg/kg citral exhibited improved scar tissue progression in obese animals, with reduced MMP-9 activity and modulation of MMP-2 activation.

Correlation between CXCR4 and MMP-2 Expression with T Stage in Clear Cell Renal Cell Carcinoma

Clear cell renal cell carcinoma (ccRCC) is the most common type of kidney cancer with high mortality. Escalation of T stage associates with worse survival. Proper and significant biomarker examination necessary to determine the predictive factor and the opportunity of targeted therapy as well. Upregulated CXCR4 expression on cancer cell promotes its aggressive growth, dissemination, metastasis, and conventional therapy resistance. Overexpression of CXCR4 may induce MMP-2 activity on tumor cell, an enzyme that degrade extracellular matrix and basal membrane particularly, which associated with tumor invasion and metastasis. Those biomarkers activity might be advantaged under hypoxic microenvironment in ccRCC. This study aimed to analyze the role and correlation of CXCR4 and MMP-2 expression with various T stages in ccRCC. Analytic observational study with cross-sectional approach was conducted on 43 formalin-fixed paraffin-embedded tissue of patients diagnosed as ccRCC in Anatomical Pathology Laboratory of Dr. Soetomo Hospital throughout January 2015 until December 2020. CXCR4 and MMP-2 expression were evaluated by immunohistochemistry. Statistical analysis with Kruskal-Wallis and Spearman test were utilized to analyze the expression difference and correlation. Difference of both CXCR4 (p=0.016) and MMP-2 (p=0.029) expression were obtained on various T stages. There was a significant positive correlation between CXCR4 expression (p=0.001) and MMP-2 expression (p=0.002) with T stage, also between CXCR4 with MMP-2 expression in various T stage of ccRCC (p=0.000). In conclusion, this study exhibited that CXCR4 and MMP-2 expression showed synergistic and positive correlation with T stage and might be considered as the basis of targeted therapy in ccRCC.

A Comparative Analysis of Treatment-Related Changes in the Diagnostic Biomarker Active Metalloproteinase-8 Levels in Patients with Periodontitis.

Previous studies have revealed the potential diagnostic utility of aMMP-8, an active form of MMP-8, in periodontal and peri-implant diseases. While non-invasive point-of-care (PoC) chairside aMMP-8 tests have shown promise in this regard, there is a dearth of literature on the evaluation of treatment response using these tests. The present study aimed to investigate treatment-related changes in aMMP-8 levels in individuals with Stage III/IV-Grade C periodontitis compared to a healthy control group, using a quantitative chairside PoC aMMP-8 test, and to determine its correlation with clinical parameters. The study included 27 adult patients (13 smoker, 14 non-smoker) with stage III/IV-grade C periodontitis and 25 healthy adult subjects. Clinical periodontal measurements, real-time PoC aMMP-8, IFMA aMMP-8, and Western immunoblot analyses were performed before and 1 month after anti-infective scaling and root planing periodontal treatment. Time 0 measurements were taken from the healthy control group to test the consistency of the diagnostic test. Both PoC aMMP-8 and IFMA aMMP-8 tests showed a statistically significant decrease in aMMP-8 levels and improvement in periodontal clinical parameters following treatment (p < 0.05). The PoC aMMP-8 test had high diagnostic sensitivity (85.2%) and specificity (100.0%) for periodontitis and was not affected by smoking (p > 0.05). Treatment also reduced MMP-8 immunoreactivity and activation as demonstrated by Western immunoblot analysis. The PoC aMMP-8 test shows promise as a useful tool for the real-time diagnosis and monitoring of periodontal therapy.