Markers Matrix Metalloproteinase Research Articles

Anti-lung cancer targets of ellagic acid and biological interaction with a blood carrier protein

Ellagic acid (C14H6O8, 4,4′,5,5′6,6′-hexahydroxydiphenic acid 2,6,2′6′-dilactone) as a metabolite polyphenol compound has shown potential anticancer activities. However, its potential interaction affinity with plasma carrier proteins and antimetastatic activities are still unknown. Therefore, in this study, the anticancer and antimetastatic activities of ellagic acid in H358 metastatic lung cancer cells were assessed by MTT, qRT-PCR and western blotting. Then, the interaction of ellagic acid with human transferrin (HTf) as a drug carrier protein was measured. The results showed that ellagic acid-triggered cytotoxicity in H358 metastatic cells with an IC50 concentration of 7.18 µM. was mediated by overexpression of Bax/Bcl-2 ratio and caspase-3 mRNA as well as caspase-3 activity. Additionally, after being incubated with ellagic acid, the mRNA and protein expression levels of vascular endothelial growth factor (VEGF), a hallmark of angiogenesis, and the metastasis markers matrix metalloproteinase-2 (MMP-2) and MMP-9 were noticeably reduced. In protein–ligand interaction section, we found that one molecule of ellagic acid potentially binds to one molecule of HTf mediated by the involvement of hydrogen bonds and van der Waals forces, supported by molecular docking simulations. In conclusion, our novel data suggested that ellagic acid can show promising antimetastatic activity against H358 lung cancer cells as well as ideal binding affinity to a drug carrier protein model.

LncRNA SNHG11 promotes malignant progression of colorectal cancer cells through the PI3K/Akt/mTOR signaling pathway

Objective: To investigate the effects of lncRNA SNHG11 on proliferation, migration, invasion and apoptosis of colorectal cancer cancer cells and possible mechanisms. Methods: qRT-PCR was performed to detect the expression level of lncRNA SNHG11 in colorectal cancer tissues and its related cell lines. The correlation between SNHG11 expression and clinical prognosis of patients was assessed by bioinformatics techniques. Cultured CRC cell lines were transfected with shCtrl (shCtrl group), shSNHG11#1 (shSNHG11#1 group), shSNHG11#2 (shSNHG11#2 group), Control cDNA (Control cDNA group), and SNHG11 cDNA (SNHG11 cDNA), respectively. Thiazolyl blue (MTT), clone formation assay, Transwell assay, cell scratch assay, and flow cytometry were used to detect the proliferation, migration, invasion, and apoptosis of CRC cells in each group. Western protein blotting was used to detect the expression of relevant proteins in each group, and the effect of lncRNA SNHG11 knockdown on the growth of tumour cells in vivo was analysed by nude mice tumouring assay. Phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signalling pathway inhibitor LY294002 was used for rescue experiments. Results: The expression of lncRNA SNHG11 was significantly higher in colorectal cancer cells and tissues than in normal tissues (P<0.05). Survival analysis showed that the expression level of SNHG11 was not statistically associated with CRC survival (P>0.05). shSNHG11#2 group compared with shCtrl group. MTT OD490/570 values decreased, the number of CRC cell clones decreased, the number of Transwell cells decreased, the area of cell scratch decreased, and the apoptosis rate increased (P<0.05). The mesenchymal markers matrix metalloproteinase (MMP9), N-cadherin and vimentin were significantly reduced, and the expression of the epithelial marker E-cadherin was upregulated. The expression of anti-apoptotic proteins Bcl-2 and Bcl-xl was decreased, and the expression of pro-apoptotic protein Bax was increased (P<0.05).In vivo experiments showed that lncRNA SNHG11 knockdown inhibited the growth of colorectal cancer cells, and the expression of Ki67 was reduced in tumours (P<0.05). LncRNA SNHG11 knockdown inhibited the expression of p-PI3K, p-Akt and p-mTOR.The PI3K/Akt/mTOR signaling pathway inhibitor LY294002 was able to restore the malignant cytological progression of colorectal cancer cells induced by the overexpression of lncRNA SNHG11. Conclusions: LncRNA SNHG11 is highly expressed in colorectal cancer. lncRNA SNHG11 can promote the malignant progression of colorectal cancer cells by regulating the PI3K/Akt/mTOR signaling pathway, and this finding provides a new theoretical basis for targeted therapy of colorectal cancer.

Клініко-інструментальні, цитологічні та інфламатометричні маркери ремоделінгу дихальних шляхів у хворих на бронхіальну астму дітей

Purpose - to investigate the clinical-instrumental, cytological and inflammatory characteristics of the disease in children with bronchial asthma, taking into account the content of the remodeling marker matrix metalloproteinase-9 (MMP-9) in the sputum to optimize the timely detection of structural remodeling of the bronchi. Materials and methods. A cohort of 99 children with BA was examined by the method of random sampling in the conditions of the Pulmonary Allergology Department of the RMCE CRCCH. All children underwent a complex clinical and paraclinical examination with the determination of the cytological composition of sputum, the determination of bronchial hyperreactivity and their hypersensitivity in a sample with serial dilution of histamine, the study of the concentration of metabolites of nitrogen monoxide in peripheral blood and condensate of pulmonary expiratory air. Two clinical groups were formed. The Group I included 55 patients in whom the content of MMP-9 in the sputum supernatant did not reach the level of 5.4 ng/ml, and the II group was formed by 44 patients with MMP-9 content ≥5.4 ng/ml. The main clinical characteristics of the observation groups were comparable. Results. It was established that the risk of a clinically more severe exacerbation of the disease is twice as high in children with an increased content of MMP-9 in the sputum (relative risk - 1.4, odds ratio - 2.0) with the preservation of this trend during 5 days of inpatient treatment. The work also shows that in patients with an increased content of MMP-9 in sputum, the concentration of FeNO more than 22.0 μmol/l occurs twice as often as in the first clinical group (pφ&lt;0.05). This finding reflects the association of the processes of allergic inflammation of the respiratory tract and their remodeling. The results of the PC20H bronchoprovocation test indicate statistically significantly lower levels of the provocation dose and provocation concentration of histamine, which corresponds to significant bronchial hypersensitivity, the chances of which increase against the background of the accumulation of MMP-9 in sputum: 27.7 times for PC20H (FEV1) &lt;3.7mg/ml and 48.8 times for PD20H (FEV1) &lt;0.8mg. Conclusions. An increase in the content of MMP-9 in the supernatant fluid of sputum was associated with a more severe course of exacerbations of bronchial asthma in children, an increase in the content of metabolites of nitrogen monoxide in the blood and lung expiratory air, more pronounced signs of bronchial hypersensitivity. The research was carried out in accordance with the principles of the Helsinki Declaration. The study protocol was approved by the Local Ethics Committee of participating institution. The informed consent of the patient was obtained for conducting the studies. No conflict of interests was declared by the authors.

CAMKK2 is upregulated in primary human osteoarthritis and its inhibition protects against chondrocyte apoptosis

To investigate the role of calcium/calmodulin-dependent protein kinase kinase 2 (CAMKK2) in human osteoarthritis. Paired osteochondral plugs and articular chondrocytes were isolated from the relatively healthier (intact) and damaged portions of human femoral heads collected from patients undergoing total hip arthroplasty for primary osteoarthritis (OA). Cartilage from femoral plugs were either flash frozen for gene expression analysis or histology and immunohistochemistry. Chondrocyte apoptosis in the presence or absence of CAMKK2 inhibition was measured using flow cytometry. CAMKK2 overexpression and knockdown in articular chondrocytes were achieved via Lentivirus- and siRNA-mediated approaches respectively, and their effect on pro-apoptotic and cartilage catabolic mechanisms was assessed by immunoblotting. CAMKK2 mRNA and protein levels were elevated in articular chondrocytes from human OA cartilage compared to paired healthier intact samples. This increase was associated with elevated catabolic marker matrix metalloproteinase 13 (MMP-13), and diminished anabolic markers aggrecan (ACAN) and type II collagen (COL2A1) levels. OA chondrocytes displayed enhanced apoptosis, which was suppressed following pharmacological inhibition of CAMKK2. Levels of MMP13, pSTAT3, and the pro-apoptotic marker BAX became elevated when CAMKK2, but not its kinase-defective mutant was overexpressed, whereas knockdown of the kinase decreased the levels of these proteins. CAMKK2 is upregulated in human OA cartilage and is associated with elevated levels of pro-apoptotic and catabolic proteins. Inhibition or knockdown of CAMKK2 led to decreased chondrocyte apoptosis and catabolic protein levels, whereas its overexpression elevated them. CAMKK2 may be a therapeutic target to prevent or mitigate human OA.

Evaluation of Fibrosis Markers as a Potential Method for Diagnosing Non-Obstructive Coronary Artery Disease in Patients with Stable Coronary Artery Disease

Aim. To study the levels of fibrosis markers in patients with stable coronary artery disease (CAD) and various types of coronary artery (CA) lesions (obstructive and non-obstructive), to identify possible differences for diagnosing the types of coronary obstruction.Material and methods. The observational study included three groups of patients: with non-obstructive (main group, coronary artery stenosis &lt;50%; n=20) and obstructive (comparison group, hemodynamically significant coronary artery stenosis according to the results of coronary angiography; n=20) CAD and healthy volunteers (control group; n=40). Transforming growth factor beta 1 (TGF-β1) and matrix metalloproteinase 9 (MMP-9) levels were measured in plasma by enzyme immunoassay. According to the results of echocardiography, all patients included in the study were divided into four groups depending on the type of myocardial remodeling.Results. TGF-β1 levels were significantly higher in patients with obstructive CAD (p=0.008) than in patients with non-obstructive CAD and healthy volunteers (p &lt;0.001). There were no significant differences between the main and control groups (p&gt;0.05). There were no statistically significant differences in TGF-β1 levels depending on the type of left ventricular remodeling (p=0.139). The maximum level of MMP-9 was in the group with obstructive coronary disease and significantly differed from the main group (p &lt;0.001) and the control group (p=0.04).Conclusio. The maximum levels of TGF-β1 and MMP-9 were found in the group with obstructive coronary artery disease. The levels of these biomarkers in the main group were statistically different from the values obtained in the control group. Thus, considering the pathogenesis of the development of non-obstructive CAD, the use of fibrosis markers TGF-β1 and MMP-9 may be promising for diagnosing the severity of CA obstruction.

Co-expression of Matrix Metalloproteinase 9 (MMP9) and Motility-related Protein-1 (MRP-1/CD9) in Human Breast Cancer

Background: Controversial evidence exists regarding the metastasis-suppressor or promoting activity of motility-related protein-1 (MRP-1/CD9) in breast cancer (BC). Objectives: we aimed at investigating the possible correlation of CD9 with a metastasis marker matrix metalloproteinase 9 (MMP9) in human BC tissues. Methods: A total of 19 BC and 5 normal breast tissues were analyzed. The mRNA expression of CD9 and MMP9 was measured by quantitative PCR. The correlation of genes’ expression with each other and clinicopathological features (i. e. tumor pathology, grade, estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER-2), P53, and KI67) was tested by relevant statistical analysis methods. Results: There was no significant difference between patients with BC and the control group regarding the expression of MMP9 (P = 0.394) and CD9 (P = 0.887). A significant strong positive correlation was observed between CD9 and MMP-9 expressions (r = 0.761 and P = 0.0002). The MMP9 expression was significantly higher in ER or PR positive compared to ER or PR negative tumors (P = 0.046). Conclusions: Given the strong correlation with MMP9, it seems that CD9 might have a role in metastasis in human BC. However, much more studies are required to further support this hypothesis.

Histone Deacetylase 3 Inhibition Decreases Cerebral Edema and Protects the Blood–Brain Barrier After Stroke

We have previously shown that selective inhibition of histone deacetylase 3 (HDAC3) decreases infarct volume and improves long-term functional outcomes after stroke. In this study, we examined the effects of HDAC3 inhibition on cerebral edema and blood–brain barrier (BBB) leakage and explored its underlying mechanisms. Adult male Wistar rats were subjected to 2-h middle cerebral artery occlusion (MCAO) and randomly treated i.p. with either vehicle or a selective HDAC3 inhibitor (RGFP966) at 2 and 24 h after stroke. Modified neurological severity scores (mNSS) were calculated at 2 h, 1 day, and 3 days. H&E, Evans blue dye (EBD) assay, and fluorescein isothiocyanate (FITC)-dextran were employed to assess cerebral edema and BBB leakage. Western blot for matrix metalloproteinase-9 (MMP9), MMP-9 zymography, and immunostaining for HDAC3, GFAP, Iba-1, albumin, aquaporin-4, claudin-5, ZO-1, and NF-kB were performed. Early RGFP966 administration decreased cerebral edema (p = 0.002) and BBB leakage, as measured by EBD assay, FITC-dextran, and albumin extravasation (p < 0.01). RGFP966 significantly increased tight junction proteins (claudin-5 and ZO-1) in the peri-infarct area. RGFP966 also significantly decreased HDAC3 in GFAP + astrocytes, which correlated with better mNSS (r = 0.67, p = 0.03) and decreased cerebral edema (r = 0.64, p = 0.04). RGFP966 decreased aquaporin-4 in GFAP + astrocytes (p = 0.002), as well as, the inflammatory markers Iba-1, NF-kB, and MMP9 in the ischemic brain (p < 0.05). Early HDAC3 inhibition decreases cerebral edema and BBB leakage. BBB protection by RGFP966 is mediated in part by the upregulation of tight junction proteins, downregulation of aquaporin-4 and HDAC3 in astrocytes, and decreased neuroinflammation.

A Novel Magnetoelastic Biosensor With Flexible TbDyFe Film as Sensing Materials for Osteoarthritis Marker MMP-3 Detection

A new design of flexible magnetoelastic (ME) biosensor based on TbDyFe/polystyrene-poly (ethylene-butylene)-polystyrene block copolymer (SEBS) film for the osteoarthritis (OA) marker matrix metalloproteinase 3 (MMP-3) detection is presented. This article studied a novel flexible ME film to replace the traditional rigid substrate for biosensors. The magnetostrictive material of TbDyFe was mixed homogeneously with paraffin solution to fabricate TbDyFe fluid, which was reacted at high temperature with SEBS in a certain ratio to prepare the film. TbDyFe/SEBS films were used as substrates for the immobilization of MMP-3 antibodies. Specific binding of antibodies to MMP-3 antigens generated compressive stress on the substrate surface, which in turn led to changes in magnetic permeability and impedance. To improve the sensitivity of the biosensor, the TbDyFe doping concentration was optimized and the magnetic response performance of the TbDyFe/SEBS composite film was measured. The results show that this assay is capable of detecting concentrations of MMP-3 between 0.76 and 10000 ng <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\cdot $ </tex-math></inline-formula> mL−1 with a detection limit of 0.76 ng <inline-formula xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink"> <tex-math notation="LaTeX">$\cdot $ </tex-math></inline-formula> mL−1, covering the range of clinical tests required for the diagnosis of OA. This novel flexible ME biosensor provides a viable method for the diagnosis and monitoring of OA.

LncRNA SNHG15 as a ceRNA modulates Osteoclast Differentiation, Proliferation, and Metastasis by Sponging miR-381-3p/NEK2 Axis.

Background A growing number of studies have shown that long noncoding RNAs play an important role in osteoclast differentiation. However, there are few studies on the roles of lncRNA small nucleolar RNA host gene 15 (SNHG15) in osteoclast differentiation. Methods The expressions of SNHG15, miR-381-3p, and never in mitosis-related kinase 2 (NEK2) mRNA were detected by real-time quantitative polymerase chain reaction (RT-qPCR); Western blot detected NEK2 and osteoclast markers (Cathepsin K, CTSK), matrix metalloproteinase 9 (MMP9), nuclear factor of activated T cell 2 (NFAT2), and tartrate-resistant acid phosphatase (TRAP) protein levels; cell proliferation was detected by Cell Counting Kit-8 (CCK-8), and the formation of osteoclasts was observed by TRAP staining; the F-actin skeleton was stained with tetramethylrhodamine isothiocyanate (TRITC) phalloidin; cell migration rate was detected by Transwell; dual-luciferase reporter gene assay and RNA-binding protein immunoprecipitation (RIP) assay verified the targeting relationship between miR-381-3p, SNHG15, and NEK2. Results The expression of SNHG15 was increased in THP-1 cells stimulated by macrophage colony-stimulating factor (M-CSF)/receptor activator of nuclear factor-kappa B ligand (RANKL). Overexpression of SNHG15 significantly promoted the proliferation, migration, osteoclast differentiation, and expression of osteoclast markers CTSK, MMP9, NFAT2, and TRAP of THP-1 cells induced by M-CSF/RANKL. Knockdown of SNHG15 reversed this effect. Overexpression of SNHG15 downregulated the inhibitory effect of overexpression of miR-381-3p on the proliferation, migration, and differentiation of THP-1 cells induced by M-CSF/RANKL. Knockdown of miR-381-3p reversed the inhibitory effect of knockdown of NEK2 on the proliferation, migration, and differentiation of THP-1 cells induced by M-CSF/RANKL. Conclusion SNHG15 acted as a ceRNA promoted the proliferation, migration, and differentiation of THP-1 cells induced by M-CSF/RANKL through sponging miR-381-3p to promote the expression of NEK2.

Epidermal Growth Factor Is Increased in Conjunctival Malignant Melanoma.

Conjunctival malignant melanoma (CMM) is a rare, but very aggressive tumor with a high metastasis rate. Not much is known about the CMM metastasis mechanisms. So far, epidermal growth factor (EGF) and its receptor (EGF-R) as well as macrophages and matrix metalloproteinase 9 (MMP-9) have been reported to lead to metastasis by epithelial-mesenchymal-transition and tumor migration in different solid tumors. Therefore, we evaluated whether EGF and EGF-R, CD68 and MMP-9 are altered in CMM samples in comparison to conjunctival nevi and healthy conjunctiva. EGF, EGF-R, the macrophage marker CD68 and MMP-9 expression were analyzed in human conjunctival melanoma (CMM, n=16), human conjunctival nevi (n=13) and disease-free human conjunctiva (controls, n=14) by immunohistology. Staining of each sample was evaluated using a standardized score ranging from negative (0) to triple positive (3). The groups were then compared by ANOVA, followed by Tukey's post-hoc test. A statistically significant increase of EGF was seen in CMM samples in comparison to conjunctival nevi (p=0.03). In contrast, no statistically significant differences in EGF-R expression were noted between the three groups. A statistically significant increase of CD68 was only seen in conjunctival nevi compared to controls (p=0.04). MMP-9 expression was similar in all groups. In CMM, the study data demonstrated an up-regulation of EGF in comparison to conjunctival nevi. Hence, EGF might promote proliferation of CMM cells and induce the epithelial-mesenchymal transition. Therefore, our data suggest that an interplay between EGF and CMM might have a critical role in the developing CMM tumors and metastasis.

Expression of Inflammatory Markers RANK, MMP-9 and PTHrP in Chronic Apical Periodontitis from People Living with HIV Undergoing Antiretroviral Therapy.

To compare the expression of the receptor activator of nuclear factor-kappa B (RANK), matrix metalloproteinase 9 (MMP-9) and parathyroid hormone-related protein (PTHrP) in primary chronic apical periodontitis lesions (CAPLs) between people living with HIV (PLWHIV) undergoing antiretroviral therapy (ART) and HIV- individuals, 32 CAPLs (16 lesions from each group) were submitted to histopathological and immunohistochemical analyses and compared between groups. The majority of the PLWHIV group had undetectable plasma viral loads (n = 13; 81.3%). The means of TCD4+ lymphocytes, exposure to HIV-1 and the time of the use of ART were 542.1 cells/mm3 (SD = 256.4), 6.3 years (SD = 2.9) and 5.0 years (SD = 2.5), respectively. Of all variables studied, only histopathological diagnosis showed a significant difference between groups (LWHIV: granuloma n = 11 (68.0%); cyst n = 5 (31.2%); HIV-: granuloma n = 15 (93.8%); cyst n = 1 (6.2%); p = 0.015). When comparing the expressions of the three inflammatory markers between the groups, no significant differences were seen. There was no difference in the expression of RANK, PTHrP and MMP-9 in primary chronic apical periodontitis lesions between PLWHIV under ART and HIV- individuals.

Interleukin (IL)-6: A Friend or Foe of Pregnancy and Parturition? Evidence From Functional Studies in Fetal Membrane Cells.

ObjectiveProtection of the fetus within the amniotic sac is primarily attained by remodeling fetal membrane (amniochorion) cells through cyclic epithelial to mesenchymal and mesenchymal to epithelial (EMT and MET) transitions. Endocrine and paracrine factors regulate EMT and MET during pregnancy. At term, increased oxidative stress forces a terminal state of EMT and inflammation, predisposing to membrane weakening and rupture. IL-6 is a constitutively expressed cytokine during gestation, but it is elevated in term and preterm births. Therefore, we tested the hypothesis that IL-6 can determine the fate of amnion membrane cells and that pathologic levels of IL-6 can cause a terminal state of EMT and inflammation, leading to adverse pregnancy outcomes.MethodsPrimary amnion epithelial cells (AECs) were treated with recombinant IL-6 (330, 1,650, 3,330, and 16,000 pg/ml) for 48 h (N = 5). IL-6-induced cell senescence (aging), cell death (apoptosis and necrosis), and cell cycle changes were studied using flow cytometry. Cellular transitions were determined by immunocytochemistry and western blot analysis, while IL-6 signaling (activation of signaling kinases) was measured by immunoassay. Inflammatory marker matrix metalloproteinase (MMP9) and granulocyte-macrophage colony-stimulating factor (GM-CSF) concentrations were measured using a Fluorokine E assay and ELISA, respectively. Amniotic membranes collected on gestational day (D) 12 and D18 from IL-6 knockout (KO) and control C57BL/6 mice (N = 3 each) were used to determine the impact of IL-6 on cell transitions. Fold changes were measured based on the mean of each group.ResultsIL-6 treatment of AECs at physiologic or pathologic doses increased JNK and p38MAPK activation; however, the activation of signals did not cause changes in AEC cell cycle, cellular senescence, apoptosis, necrosis, cellular transitions, or inflammation (MMP9 and GM-CSF) compared to control. EMT markers were higher on D18 compared to D12 regardless of IL-6 status in the mouse amniotic sac.ConclusionPhysiologic and pathologic concentrations of IL-6 did not cause amnion cell aging, cell death, cellular transitions, or inflammation. IL-6 may function to maintain cellular homeostasis throughout gestation in fetal membrane cells. Although IL-6 is a good biomarker for adverse pregnancies, it is not an indicator of an underlying pathological mechanism in membrane cells.

Tumour-associated macrophages mediate the invasion and metastasis of bladder cancer cells through CXCL8.

Tumour-associated macrophages (TAMs) are associated with both the progression and poor prognosis of a variety of solid tumours. This study aimed to investigate and clarify the tumour-promoting role of CXCL8 secreted by TAMs in the urothelial carcinoma microenvironment of the bladder. Immunohistochemistry (n = 55) was used to detect Chemokine (C-X-C motif) ligand 8 (CXCL8), CD163 (a TAM marker), Matrixmetalloproteinase-9 (MMP-9), vascular endothelial growth factor (VEGF), and E-cadherin in cancerous and adjacent tissues of bladder cancer patients. TAMs-like PBM (peripheral blood mononuclear)-derived macrophages were developed using in vitro experiments. T24, 5637, and UM-UC-3 were treated with conditioned medium (CM) for the experimental intervention group, without CM for the blank control group, and with CM and an anti-CXCL8 neutralizing antibody for the experimental control group, respectively. The immunohistochemical study showed that the expression of CXCL8 was significantly upregulated as the number of infiltrating TAMs increased in the tumour tissues. A high expression of CXCL8 significantly correlated with an increase in the expression of MMP-9 and VEGF and a decrease in expression of E-cadherin in the microenvironment. This revealed that TAM-derived CXCL8 is highly associated with bladder cancer migration, invasion, and angiogenesis. The concentration of CXCL8 was significantly higher in CM collected from TAM-like PBM-derived macrophages than that from THP-1 cells. In subsequent in vitro experiments, we found that CM derived from TAM-like PBM-derived macrophages can also increase the migration rate, invasiveness, and pro-angiogenic properties of tumour cells. Additionally, the effect of CXCL8 was significantly diminished by the addition of an anti-CXCL8 neutralizing antibody to CM. The infiltration of TAMs in the tumour microenvironment leads to the elevation of CXCL8, which in turn promotes the secretion of MMP-9, VEGF, and E-cadherin by bladder cancer cells. This alters the migration, invasion, and pro-angiogenic capacity of bladder cancer cells and accelerates cancer progression.

HER4 promotes the growth and metastasis of osteosarcoma via the PI3K/AKT pathway.

Osteosarcoma is the most common primary malignant bone tumor, which occurs in adolescents. As reported by our previous studies, HER4 indicates a poor prognosis of primary osteosarcoma. However, its mechanisms in the pathogenesis of osteosarcoma have not yet been studied. The purpose of this study was to investigate the role of HER4 in osteosarcoma and whether the PI3K/AKT pathway is involved. In this study, western blot analysis was used to investigate the expression of HER4 protein in osteosarcoma tissues and cell lines. CCK8 and transwell assays were used to detect the effects of HER4 on the proliferation, migration, and invasion of osteosarcoma cells in vitro. The effects of HER4 on the growth and metastasis of osteosarcoma in vivo were detected by tumor formation and immunofluorescence in nude mice. The role of the PI3K/AKT pathway in HER4 regulation of the growth and metastasis of osteosarcoma was examined by western blot analysis and immunofluorescence assay. We found that HER4 protein was highly expressed in clinical osteosarcoma specimens and osteosarcoma cells. HER4 markedly promoted the proliferation, migration, and invasion of osteosarcoma cells in vitro as well as the growth and metastasis of osteosarcoma in vivo. HER4 overexpression upregulated the expression of phosphorylated protein kinase B (pAKT), proliferation marker antigen Ki67, and metastasis cell marker matrix metalloproteinase 9 (MMP9). Notably, PI3K/AKT inhibitor LY294002 significantly inhibited the effects of HER4 via the downregulation of pAKT, Ki67, and MMP9. Moreover, LY294002 markedly blocked the effects of HER4-induced upregulation of tumor malignancy. The present study suggests that HER4 may promote the growth and metastasis of osteosarcoma via the PI3K/AKT pathway. The HER4/PI3K/AKT pathway could serve as a potential target for the treatment of osteosarcoma.

Wnt/β‑catenin signaling modulates piperine‑mediated antitumor effects on human osteosarcoma cells.

The plant extract piperine is used as a traditional Chinese medicine due to its anti-inflammatory effects and efficacy against numerous types of cancer. The aim of the present study was to investigate the antitumor mechanism of piperine in human osteosarcoma U2OS and 143B cell lines. The effects of piperine on cell apoptosis and invasion of human osteosarcoma cells were assessed using flow cytometry and Transwell assays. Moreover, western blotting was used to measure the effects of piperine on the protein expression levels of the metastasis markers matrix metalloproteinase-2 (MMP-2) and vascular endothelial growth factor (VEGF). In addition, the involvement of the Wnt/β-catenin signaling pathway in modulating the effects of piperine was examined via western blot analysis. The results of MTT and Transwell invasion assays indicated that piperine treatment dose-dependently reduced U2OS and 143B cell viability and invasion. Furthermore, a significant reduction was identified in MMP-2, VEGF, glycogen synthase kinase-3β and β-catenin protein expression levels, as well as the expression levels of their target proteins cyclooxygenase-2, cyclin D1 and c-myc, in U2OS cells after piperine treatment. In addition, similar results were observed in 143B cells. Therefore, the present study demonstrated the efficacy of piperine in osteosarcoma, and identified that the Wnt/β-catenin signaling pathway may modulate the antitumor effects of piperine on human U2OS and 143B cells.

Simultaneous bioimaging of MMP-2 and MMP-7 via Au-Se constructed fluorescence nanoprobe

Au-Se bond strategy has been applied to construct fluorescence nanoprobe that is being used to simultaneously dual-image the tumor markers matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-7 (MMP-7) in vitro. The two Se-terminating ligand peptide chains respectively labeled with fluorescein isothiocyanate (FITC) and 5-Carboxytetramethylrhodamine (5-TAMRA) dyes are attached to the surface of Au nanoparticles (NPs). The nanoprobe can be specifically cut off by MMP-2 and MMP-7 respectively to carry out significant enhanced fluorescence recovery for simultaneously imaging both markers. The nanoprobe not only displays high selectivity and sensitivity towards MMP-2 and MMP-7, but also has strong anti-interference stability against biothiols and enhanced fidelity for avoiding ‘false positive’ results. This approach offers new perspectives in designing tumor-related early diagnosis approaches and also provides strategies for clinical applications.

The impact of chronic endometritis on endometrial fibrosis and reproductive prognosis in patients with moderate and severe intrauterine adhesions: a prospective cohort study

To study the effect of chronic endometritis (CE) diagnosed by CD138 immunohistochemical (IHC) staining on endometrial fibrosis and reproductive prognosis in patients with moderate or severe intrauterine adhesions (IUAs). Prospective cohort study (Canadian Task Force classification II-2). University-affiliated hospital. One hundred sixty-seven women with moderate to severe IUAs. Transcervical resection of adhesions (TCRA) and endometrial biopsy were performed in all patients. According to results of IHC staining with anti-syndecan-1 antibodies to identify CD138cells, participants were classified into two groups: 78 patients with CE (CE group) and 89 women without CE (NCE group). IHC staining for fibrosis markers transforming growth factor beta 1, anti-fibrosis markers matrix metalloproteinase 9, and endometrial receptivity marker integrin alpha v beta 3 was later applied to all tissue samples. Endometrial fibrosis, endometrial receptivity, and reproductive prognosis. CE diagnosed by CD138 IHC staining has a high incidence, 46%, in moderate and severe IUAs. In the CE group, the expression of transforming growth factor beta 1 was higher than that in the NCE group, and the expression of matrix metalloproteinase 9 and alpha v beta 3 was lower than that in the NCE group. The pregnancy rate and live birth rate in the NCE group were higher than those in the CE group (42.7% vs. 31.5%, 26.9% vs. 17.9%). CE may affect the endometrial fibrosis homeostasis in IUAs. Women with CE were more likely to experience recurrence of adhesions and had poorer reproductive outcomes. NCT02744807.

Effect of microfragmented adipose tissue on osteoarthritic synovial macrophage factors.

Cell-based therapies using adipose-derived mesenchymal stromal cells (ADMSCs) have shown promising results for the treatment of osteoarthritis (OA). In fact, ADMSCs are now indicated as one of the most powerful cell sources through their immunomodulatory and anti-inflammatory activities. Recently, an innovative one-step closed device was developed to obtain microfragmented adipose tissue (MF) to avoid the need for good manufacturing practices for ADMSCs expansion while maintaining their regenerative potential. The aim of this study was to assess the mechanisms of action of MF and ADMSCs from MF (MF-ADMSCs) on an inflammatory cell model of OA synoviocytes. We found that MF produced low levels of inflammatory factors such as interleukin 6 (IL-6), CC-chemokine ligand 5/receptor-activated normal T-cell expressed and secreted (CCL5/RANTES), CC-chemokine ligand 2/monocyte chemoattractant protein-1 (CCL2/MCP-1), and CC-chemokine ligand 3/macrophage inflammatory protein-1α (CCL3/MIP-1α), and a higher level only of CXC-chemokine ligand 8/interleukin 8 compared with MF-ADMSCs. Matrix metalloproteinase 9 (MMP-9) degradative factor but released a lower level of its inhibitor tissue inhibitor of the metalloproteinase (TIMP-1). MF in coculture with synoviocytes significantly induced both the metabolic activity and the release of IL-6. In contrast, MF, not MF-ADMSCs, partially decreased CCL5/RANTES. Moreover, MF reduced the release of both macrophage-specific chemokines (CCL2/MCP-1 and CCL3/MIP-1α) and degradative marker MMP-9. Interestingly, MF increased TIMP-1 (the MMP-9 inhibitor) and down-modulated toll-like receptor (TLR4) receptor and key molecules of NFκB pathways. These data evidenced different effects of MF versus MF-ADMSCs on inflamed synoviocytes. MF reduced typical macrophages markers and its potentiality by switching off macrophages activity was strictly dependent on TLR4 and NFκB signaling.

Effect of remote ischemic preconditioning on left atrial remodeling and prothrombotic response after radiofrequency catheter ablation for atrial fibrillation

Radiofrequency catheter ablation (RFCA) of atrial fibrillation (AF) is known to induce left atrial remodeling and prothrombotic response. This study aimed to evaluate the effect of remote ischemic preconditioning (RIPC) on left atrial remodeling and prothrombotic response induced by RFCA of AF. Forty-four patients with drug-refractory paroxysmal AF undergoing RFCA were randomized into RIPC (four short episodes of forearm ischemia) and control groups before the procedure. Blood samples were collected before RIPC/sham RIPC, and 24 and 72hours later after the procedure. The atrial remodeling marker matrix metalloproteinase-9 (MMP-9) and endothelial damage marker von Willebrand factor (vWF) were measured using enzyme-linked immunosorbent assay. Plateletactivation was evaluated by flow cytometric measurements of the expression of platelet P-selectin (CD62P) and active glycoprotein IIb/IIIa receptor (PAC-1). The early recurrence of atrial fibrillation (ERAF) in the two groups was observed over the subsequent 3 months. RFCA resulted in a significant increase in MMP-9 and vWF in both the groups, which persisted for 72hours. However, the expression of CD62P and PAC-1 showed less increase during RFCA in either group. The RIPC group showed a lower increase in MMP-9 and vWF compared with the control group. In contrast, no significant differences were found in the trend of expression of CD62P and PAC-1 during RFCA between the two groups. The AF recurrence in the 3 months after the ablation was significantly lower in the RIPC group than in the control group. RIPC before RFCA for paroxysmal AF significantly reduces the increase in markers of left atrial remodeling and endothelial damage associated with the procedure, and results in a lower ERAF.

Study of miR-10b regulatory mechanism for epithelial-mesenchymal transition, invasion and migration in nasopharyngeal carcinoma cells.

The aim of the study was to investigate the miR-10b regulatory mechanism for epithelial-mesenchymal transition (EMT) and its effect on the proliferation and migration of nasopharyngeal carcinoma cells. RT-qPCR was used to detect the expression of miR-10b in CNE1 nasopharyngeal carcinoma cell line. The NP69 nasopharyngeal mucosal cell line was used to determine the expression of miR-10b after infection with lentivirus. The effect of miR-10b on the proliferation of NP69 was examined using cell counting kit-8. The effect of miR-10b on NP69 migration was examined using scratch assay. Western blot analysis was used to detect the effects of miR-10b on the expression of epithelial cell markers E-cadherin and β-catenin and mesenchymal cell markers fibronectin, N-cadherin, vimentin and matrix metalloproteinase-9 (MMP-9). The present study showed that miR-10b was highly expressed in CNE1 cells. The stable expression of miR-10b promoted the proliferation and migration of NP69 cells, downregulated the expression of epithelial cell markers E-cadherin and β-catenin, and upregulated the expression of mesenchymal cell markers fibronectin, N-cadherin, vimentin and MMP-9 resulting in cell EMT. In conclusion, miR-10b promotes the proliferation and migration of nasopharyngeal carcinoma cells, and induces EMT in nasopharyngeal carcinoma cells, thereby having the potential to become a new target for the treatment of nasopharyngeal carcinoma.