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OBJECTIVE@#To explore the changes of Ⅻ antithrombin (FⅫa-AT), thrombospondin-1 (TSP-1), and lupus anticoagulant (LA) ratio in the peripheral blood factor of patients with systemic lupus erythematosus (SLE) and the clinical value of combined diagnosis of thrombotic events.@*METHODS@#A total of 133 SLE patients treated in Xingtai People's Hospital were selected and divided into simple SLE group (105 cases) and SLE complicated with thrombosis group (28 cases) according to whether thrombotic events occurred, and 102 cases of healthy people in the same period were selected as control. The clinical data of the 3 groups, the level of peripheral blood FⅫa-AT, TSP-1, and LA ratio were compared, the relationship between each peripheral blood index and SLE disease activity index (SLEDAI) score were analyzed. The influencing factors of thrombotic events in SLE patients were analyzed, and the value of each peripheral blood index in the diagnosis of SLE complicated with thrombotic events were evaluated.@*RESULTS@#The proportion of the patients with age ≥60 year, hypertension, and smoking history in SLE complicated with thrombosis group was higher than those in simple SLE group and control group (P<0.05). The SLEDAI score, peripheral blood FⅫa-AT, TSP-1, LA ratio levels of the patients in SLE complicated with thrombosis group were significantly higher than those in simple SLE group and control group, and the simple SLE group was significantly higher than the control group (P<0.05). FⅫa-AT, TSP-1, LA ratio in peripheral blood of SLE patients were positively correlated with SLEDAI score (r=0.663, 0.578 and 0.625). Age, blood pressure, smoking history, peripheral blood FⅫa-AT, TSP-1, LA ratio were the important influencing factors of thrombotic events in SLE patients (P<0.05). The AUC diagnosed by the FⅫa-AT, TSP-1, and LA ratio in peripheral blood was 0.881, the 95% CI was 0.813-0.931, the sensitivity was 82.14%, and the specificity was 91.43%, which was superior to each index alone (P<0.05).@*CONCLUSION@#Peripheral blood FⅫa-AT, TSP-1, LA ratio level changes in SLE patients are significantly related to disease activity, and the combined diagnosis of thrombotic events is more reliable.
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Humanos , Lúpus Eritematoso Sistêmico/complicações , Fatores de Risco , Trombose/etiologia , Trombospondina 1RESUMO
Fibrosis caused by the increase in extracellular matrix in cardiac fibroblasts plays an important role in the occurrence and development of atrial fibrillation (AF). The aim of this study was to investigate the role of hsa-miR-4443 in AF, human cardiac fibroblast (HCFB) proliferation, and extracellular matrix remodeling. TaqMan Stem-loop miRNA assay was used to measure hsa-miR-4443 expression in patients with persistent AF (n=123) and healthy controls (n=100). Patients with AF were confirmed to have atrial fibrosis by late gadolinium enhancement. At the cellular level, after hsa-miR-4443 mimic and inhibitor were transfected with HCFBs, proliferation, apoptosis, migration, and invasion were analyzed. Lastly, hsa-miR-4443-targeted gene and transforming growth factor (TGF)-β1/α-SMA/collagen pathway were evaluated by dual-luciferase reporter assay and western blot, respectively. In patients with AF, hsa-miR-4443 decreased significantly and collagen metabolism level increased significantly. Logistic regression analysis showed that low hsa-miR-4443 level was a risk factor of AF (P<0.001). The receiver operating characteristic curve revealed that hsa-miR-4443 was useful for predicting AF (area under the curve: 0.828, sensitivity: 0.71, specificity: 0.78, P<0.001). In HCFBs, hsa-miR-4443 targeted thrombospondin-1 (THBS1) and downregulated TGF-β1/α-SMA/collagen pathway. The inhibition of hsa-miR-4443 expression promoted HCFB proliferation, migration, invasion, myofibroblast differentiation, and collagen production. The significant reduction of hsa-miR-4443 can be used as a biomarker for AF. hsa-miR-4443 protected AF by targeting THBS1 and regulated TGF-β1/α-SMA/collagen pathway to inhibit HCFB proliferation and collagen synthesis.
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Humanos , Fibrilação Atrial , MicroRNAs/genética , Fibrose , Colágeno , Meios de Contraste , Trombospondina 1/genética , Proliferação de Células , Fator de Crescimento Transformador beta1 , Fibroblastos , GadolínioRESUMO
PURPOSE: Polycystic ovary syndrome (PCOS) is a gynecological endocrine disorder that is characterized by disturbances in ovarian blood flow and angiogenesis. The aim of this study was to determine the association of vascular endothelial growth factor (VEGF) and thrombospondin-1 (TSP-1) serum levels with the body mass index (BMI) in patients with PCOS compared with healthy subjects. METHODS: The study was conducted with 80 subjects in 3 PCOS groups, including normal weight, overweight, and obese PCOS groups, and a control group of healthy subjects (n=20). The participants in all groups completed a questionnaire comprising sociodemographic and obstetric questions. The PCOS diagnosis in the study subjects was confirmed based on the Rotterdam criteria, BMI was determined according to the World Health Organization guidelines, and the lipid accumulation product index was calculated for all groups. Venous blood samples were collected from all participants after fasting to measure the serum levels of fasting blood glucose (FBG), lipids, insulin, VEGF, TSP-1, and leptin. RESULTS: Our findings showed that the serum VEGF level was significantly higher in the normal BMI PCOS group than that in the control group (P=0.03), and the TSP-1 level was significantly lower in the obese PCOS group than that in the control group (P=0.04). CONCLUSIONS: Our study demonstrated that alterations in VEGF and TSP-1 concentrations are dependent on BMI. Because abnormal ovarian angiogenesis is considered to be the main feature of PCOS, the study of ovarian angiogenic imbalance is proposed as a new tool for PCOS diagnosis and management.
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Humanos , Glicemia , Índice de Massa Corporal , Estudos de Casos e Controles , Diagnóstico , Jejum , Voluntários Saudáveis , Insulina , Leptina , Produto da Acumulação Lipídica , Sobrepeso , Síndrome do Ovário Policístico , Trombospondina 1 , Fator A de Crescimento do Endotélio Vascular , Organização Mundial da SaúdeRESUMO
Thrombospondin-1 (TSP-1) is widely distributed in human tissues and is important in inhibiting angiogenesis.It also occupies an indispensable position in the formation, growth, differentiation, and metastasis of tumors in different tissues.TSP-1 plays an important role in the occurrence and development of various types of tumors. The inhibitory effect of TSP-1 on the angiogenesis and tumor development of oral and maxillofacial malignant tumors has been demonstrated in recent years. This paper reviews the findings and progress of TSP-1 research involving all kinds of tumors as well as oral and maxillofacial malignancies.
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Humanos , Neoplasias , Neovascularização Patológica , Trombospondina 1RESUMO
Sphingosylphosphorylcholine (SPC) is one of the bioactive phospholipids that has many cellular functions such as cell migration, adhesion, proliferation, angiogenesis, and Ca²⁺ signaling. Recent studies have reported that SPC induces invasion of breast cancer cells via matrix metalloproteinase-3 (MMP-3) secretion leading to WNT activation. Thrombospondin-1 (TSP-1) is a matricellular and calcium-binding protein that binds to a wide variety of integrin and non-integrin cell surface receptors. It regulates cell proliferation, migration, and apoptosis in inflammation, angiogenesis and neoplasia. TSP-1 promotes aggressive phenotype via epithelial mesenchymal transition (EMT). The relationship between SPC and TSP-1 is unclear. We found SPC induced EMT leading to mesenchymal morphology, decrease of E-cadherin expression and increases of N-cadherin and vimentin. SPC induced secretion of thrombospondin-1 (TSP-1) during SPC-induced EMT of various breast cancer cells. Gene silencing of TSP-1 suppressed SPC-induced EMT as well as migration and invasion of MCF10A cells. An extracellular signal-regulated kinase inhibitor, PD98059, significantly suppressed the secretion of TSP-1, expressions of N-cadherin and vimentin, and decrease of E-cadherin in MCF10A cells. ERK2 siRNA suppressed TSP-1 secretion and EMT. From online PROGgene V2, relapse free survival is low in patients having high TSP-1 expressed breast cancer. Taken together, we found that SPC induced EMT and TSP-1 secretion via ERK2 signaling pathway. These results suggests that SPC-induced TSP-1 might be a new target for suppression of metastasis of breast cancer cells.
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Humanos , Apoptose , Neoplasias da Mama , Caderinas , Movimento Celular , Proliferação de Células , Transição Epitelial-Mesenquimal , Inativação Gênica , Inflamação , Metástase Neoplásica , Fenótipo , Fosfolipídeos , Fosfotransferases , Receptores de Superfície Celular , Recidiva , RNA Interferente Pequeno , Trombospondina 1 , VimentinaRESUMO
Sphingosylphosphorylcholine (SPC) is one of the bioactive phospholipids that has many cellular functions such as cell migration, adhesion, proliferation, angiogenesis, and Ca²⁺ signaling. Recent studies have reported that SPC induces invasion of breast cancer cells via matrix metalloproteinase-3 (MMP-3) secretion leading to WNT activation. Thrombospondin-1 (TSP-1) is a matricellular and calcium-binding protein that binds to a wide variety of integrin and non-integrin cell surface receptors. It regulates cell proliferation, migration, and apoptosis in inflammation, angiogenesis and neoplasia. TSP-1 promotes aggressive phenotype via epithelial mesenchymal transition (EMT). The relationship between SPC and TSP-1 is unclear. We found SPC induced EMT leading to mesenchymal morphology, decrease of E-cadherin expression and increases of N-cadherin and vimentin. SPC induced secretion of thrombospondin-1 (TSP-1) during SPC-induced EMT of various breast cancer cells. Gene silencing of TSP-1 suppressed SPC-induced EMT as well as migration and invasion of MCF10A cells. An extracellular signal-regulated kinase inhibitor, PD98059, significantly suppressed the secretion of TSP-1, expressions of N-cadherin and vimentin, and decrease of E-cadherin in MCF10A cells. ERK2 siRNA suppressed TSP-1 secretion and EMT. From online PROGgene V2, relapse free survival is low in patients having high TSP-1 expressed breast cancer. Taken together, we found that SPC induced EMT and TSP-1 secretion via ERK2 signaling pathway. These results suggests that SPC-induced TSP-1 might be a new target for suppression of metastasis of breast cancer cells.
Assuntos
Humanos , Apoptose , Neoplasias da Mama , Caderinas , Movimento Celular , Proliferação de Células , Transição Epitelial-Mesenquimal , Inativação Gênica , Inflamação , Metástase Neoplásica , Fenótipo , Fosfolipídeos , Fosfotransferases , Receptores de Superfície Celular , Recidiva , RNA Interferente Pequeno , Trombospondina 1 , VimentinaRESUMO
<p><b>OBJECTIVE</b>To study the inhibitory effect and mechanism of Ganoderma lipsiense extract (GLE) on the growth of triple-negative breast cancer (TNBC) cell line MDA-MB-231-HM in a mouse model.</p><p><b>METHODS</b>The mouse model of TNBC was established by subcutaneous injection of 1.5 x 10(6) of MDA-MB-231-HM cells into BALB/c-nu mouse. Twenty successfully modeled mice were divided into the GLE group and the negative control group according to random digit table, 10 in each group. GLE (0.2 mL 100 mg/mL) was peritoneally injected to mice in the GLE group, while equal dose of normal saline was peritoneally injected to mice in the negative control group. The medication was administered once per 3 days and discontinued after 45 days. The CD34 expression was detected using immunohistochemical assay for counting microvessels. Meanwhile, expressions of thrombospondin 1 (TSP-1) and cyclin D1 were detected using immunohistochemical assay.</p><p><b>RESULTS</b>The average weight was obviously lower in the GLE group than in the negative control group [(0.33 ± 0.16) g vs (0.68 ± 0.37)g, P < 0.05]. The tumor inhibition rate was 51.4% in the GLE group. The volume of transplanted tumor was obviously lesser in the GLE group than in the negative control group (P < 0.05). Results of immunohistochemical staining showed, the microvessel density (MVD) under every field was (20.7 ± 2.1), TSP-1 positive cell count was (66.2 ± 9.2), cyclin D1 positive cell count was (33.8 ± 16.4) in the GLE group, and they were 34.0 ± 2.0, 24.0 ± 6.6, and 168.2 ± 32.6, respectively in the negative control group. There was statistical difference in all indices between the two groups (P < 0.05).</p><p><b>CONCLUSION</b>GLE could inhibit malignant proliferation of tumor cells by suppressing angiogenesis of blood vessels in tumor tissues and regulating cell cycles, thereby inhibiting TNBC.</p>
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Animais , Humanos , Camundongos , Produtos Biológicos , Farmacologia , Linhagem Celular Tumoral , Ciclina D1 , Metabolismo , Modelos Animais de Doenças , Ganoderma , Química , Camundongos Endogâmicos BALB C , Camundongos Nus , Microvasos , Transplante de Neoplasias , Neovascularização Patológica , Distribuição Aleatória , Trombospondina 1 , Metabolismo , Neoplasias de Mama Triplo Negativas , Tratamento FarmacológicoRESUMO
BACKGROUND: Wound healing is an interaction of a complex signaling cascade of cellular events, including inflammation, proliferation, and maturation. K+ channels modulate the mitogen-activated protein kinase (MAPK) signaling pathway. Here, we investigated whether K+ channel-activated MAPK signaling directs collagen synthesis and angiogenesis in wound healing. METHODS: The human skin fibroblast HS27 cell line was used to examine cell viability and collagen synthesis after potassium chloride (KCl) treatment by Cell Counting Kit-8 (CCK-8) and western blotting. To investigate whether K+ ion channels function upstream of MAPK signaling, thus affecting collagen synthesis and angiogenesis, we examined alteration of MAPK expression after treatment with KCl (channel inhibitor), NS1619 (channel activator), or kinase inhibitors. To research the effect of KCl on angiogenesis, angiogenesis-related proteins such as thrombospondin 1 (TSP1), anti-angiogenic factor, basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF), pro-angiogenic factor were assayed by western blot. RESULTS: The viability of HS27 cells was not affected by 25 mM KCl. Collagen synthesis increased dependent on time and concentration of KCl exposure. The phosphorylations of MAPK proteins such as extracellular-signal-regulated kinase (ERK) and p38 increased about 2.5-3 fold in the KCl treatment cells and were inhibited by treatment of NS1619. TSP1 expression increased by 100%, bFGF expression decreased by 40%, and there is no significant differences in the VEGF level by KCl treatment, TSP1 was inhibited by NS1619 or kinase inhibitors. CONCLUSIONS: Our results suggest that KCl may function as a therapeutic agent for wound healing in the skin through MAPK signaling mediated by the K+ ion channel.
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Humanos , Western Blotting , Contagem de Células , Linhagem Celular , Sobrevivência Celular , Colágeno , Fator 2 de Crescimento de Fibroblastos , Fibroblastos , Inflamação , Canais Iônicos , Proteínas Quinases Ativadas por Mitógeno , Fosforilação , Fosfotransferases , Canais de Potássio , Cloreto de Potássio , Proteínas Quinases , Pele , Trombospondina 1 , Fator A de Crescimento do Endotélio Vascular , CicatrizaçãoRESUMO
Angiogenesis is the fundamental biological phenomenon in the development of vertebrates and various pathophysiological process such as cancer, inflammation and wound healing. Thrombospondin-1 is a well-known anti-angiogenic molecule which is distributed in the extracellular matrix of various tissues. The second and third type I repeats of human TSP-1 have inhibitory effects on endothelial cell migration and induce angiogenesis inhibition. However the role of the first type I repeat was not elucidated. In addition, the first type I repeat of bovine TSP-1 has CSVTCG amino acid sequence which is known to have anti-angiogenic activity. In the present study, we compared the inhibition of angiogenesis to investigate the role of the first type I repeat of the human and bovine TSP-1. Matrigel was mixed with or without TSR-1 peptides and then injected into C57BL/6J mice. We compared angiogenesis inhibition activity by hemoglobin assay, microvessel density and optical density value after 7 days. Furthermore, inhibition of angiogenesis was confirmed on CAM assay by TSR-1 peptides. For in vitro angiogenesis assay, TSR-1 peptides were treated on the proliferation, migration, and tube formation assay of HUVEC. Apoptosis effect of TSR-1 peptides was confirmed by apoptosis assay kit and flow cytometry. Bovine and human TSR-1 peptides blocked neovascularization in in vivo Matrigel plug assay and CAM assay at 10 microM. Bovine TSR-1 peptides have shown stronger angiogenesis inhibition in bFGF-induced angiogenesis than human TSR-1 and CSVTCG peptides. However, all of TSR-1 peptides inhibit migration and tube formation of HUVEC in in vitro. Furthermore, these peptides also induced apoptosis of HUVEC. These results suggest that TSR-1 peptides of bovine and human TSP-1 have angiogenesis inhibition activity.
Assuntos
Animais , Humanos , Camundongos , Sequência de Aminoácidos , Apoptose , Fenômenos Biológicos , Células Endoteliais , Matriz Extracelular , Citometria de Fluxo , Inflamação , Microvasos , Peptídeos , Trombospondina 1 , Vertebrados , CicatrizaçãoRESUMO
Thrmobospondin-1 is the multifunctional protein that modulates endothelial cell and tumor cell behavior via several cell surface receptors and inhibits angiogenesis. In vitro, thrombospondin-1 alters adhesion, proliferation, motility, and survival of endothelial and cancer cells. Studies have confirmed that increased TSP-1 expression suppresses growth or metastasis of some tumors and inhibits angiogenesis. In the past three decades, inhibitors of angiogenesis have been developed as regulators target the vascular endothelial growth factor (VEGF) signaling pathway and small molecule tyrosine kinase inhibitors have been clinically approved. TSP-1 has several functional domain structures and inhibits tumor angiogenesis by engaging receptors CD36 and CD47. TSP-1 binding to CD47 dissociates it from VEGFR2, inhibiting downstream AKT activation and functional responses of endothelial cells to VEGF. Recently, macrophage phagocytosis and cytotoxic T-cell induction of tumor cells mediated by CD47-specific blocking antibodies have been proposed. These findings provide a new therapeutic paradigm for elinination of cancer cells and inhibition of angiogenesis of tumor by TSP-1.
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Anticorpos Bloqueadores , Células Endoteliais , Macrófagos , Metástase Neoplásica , Fagocitose , Proteínas Tirosina Quinases , Receptores de Superfície Celular , Linfócitos T , Trombospondina 1 , Fator A de Crescimento do Endotélio VascularRESUMO
BACKGROUND: The authors analyzed whether the promoter hypermethylation of cancer-related genes was involved in the tumorigenesis of malignant gliomas. METHODS: A total of 29 patients received surgery and histologically confirmed to have malignant gliomas from January 2000 to December 2006. The promoter methylation status of several genes, which were reported to be frequently methylated in malignant gliomas, was investigated using methylation-specific polymerase chain reaction. RESULTS: All cases of malignant gliomas represented the promoter hypermethylation in at least 2 or more genes tested. Of 29 tumors, 28 (96.55%) showed concurrent hypermethylation of 3 or more genes. Ras association domain family member 1, epithelial cadherin, O-6 methyl guanine DNA methyltransferase, thrombospondin 1, p14 and adenomatous polyposis coli were frequently methylated in high grade gliomas including glioblastomas, anaplastic astrocytomas, and anaplastic oligodendrogliomas. CONCLUSION: Aberrant hypermethylation profile was closely related with malignant gliomas suggesting that epigenetic change may play a role in the development of malignant gliomas. Two or three target genes may provide useful clues to the development of the useful prognostic as well as diagnostic assays for malignant gliomas.
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Humanos , Polipose Adenomatosa do Colo , Astrocitoma , Neoplasias Encefálicas , Carcinogênese , Ilhas de CpG , DNA , Epigenômica , Glioblastoma , Glioma , Guanina , Metilação , Oligodendroglioma , Reação em Cadeia da Polimerase , Trombospondina 1RESUMO
<p><b>OBJECTIVE</b>To observe the effect of sodium tanshinone II (A) sulfonate (STS) on Ang II -induced atrial fibroblast collagen synthesis and TGF-beta1 activation.</p><p><b>METHOD</b>Atrial fibroblasts of neonatal rats were cultured to determine the content of collagen protein. The original synthesis rate determined by the [3H]-proline incorporation method was taken as the index for myocardial fibrosis. The content of active TGF-beta1 and total TGF-beta1 in cell culture supernatants were tested and cultured by ELISA. The expression of thrombospondin-1 (TSP-1) was assessed by using Western blot.</p><p><b>RESULT</b>Ang II could significantly increase the content of atrial fibroblast collagen and the collagen synthesis rate, the TSP-1 expression and the concentration of active TGF-beta1, without any obvious change in total TGF-beta1. After the STS treatment, all of the indexes, apart from total TGF-beta1, were obviously down-regulated.</p><p><b>CONCLUSION</b>STS could decrease the secretion of Ang II -induced atrial fibroblast collagen and the synthesis rate. Its mechanism is related to the inhibition of TSP-1/TGF-beta1 pathway.</p>
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Animais , Ratos , Angiotensina II , Farmacologia , Colágeno , Fibroblastos , Biologia Celular , Metabolismo , Regulação da Expressão Gênica , Átrios do Coração , Biologia Celular , Fenantrenos , Farmacologia , Ratos Wistar , Transdução de Sinais , Trombospondina 1 , Metabolismo , Fator de Crescimento Transformador beta1 , MetabolismoRESUMO
BACKGROUND: Metronomic chemotherapy (MCT) with cyclophosphamide (Cy) and celecoxib (Cel) has therapeutic efficacy and low toxicity profile in advanced breast cancer patients (ABCP), but no reliable biomarkers of response have been found yet that allow patient selection for treatment. AIM: To investigate the potential role as biomarkers of pro‑ and antiangiogenic parameters and evaluate their response in ABCP receiving metronomic Cy 50 mg p.o./day + Cel 400 mg p.o./day. MATERIALS AND METHODS: Serum levels of vascular endothelial growth factor‑C (VEGF‑C), soluble VEGF receptors 2 and 3 (sVEGFR‑2, sVEGFR‑3), were measured at different time points in 13/15 patients included in a phase II trial of MCT with Cy+Cel. RESULTS: Serum levels of sVEGFR‑2 and sVEGFR‑3 increased significantly during treatment (P = 0.0392; P = 0.0066, respectively). VEGF‑C showed no significant modifications. Previous determinations of VEGF and TSP‑1 in the same patients were utilized. VEGF/sVEGFR‑2, VEGF/TSP‑1, and VEGF‑C/sVEGFR‑3 ratios decreased significantly along the treatment (P = 0.0092; P = 0.0072; P = 0.0141, respectively). Nonsignificant variations were observed for VEGF‑C/sVEGFR‑2 ratio. Baseline values of VEGF/sVEGFR‑2 and VEGF/TSP‑1 ratios were associated with time to progression (TTP) (P = 0.0407; P = 0.0394, respectively) meanwhile baseline VEGF was marginally significant (P = 0.0716). Patients with values lower than the 50th percentile for both ratios showed longer TTP. CONCLUSIONS: We have identified the baseline VEGF/sVEGFR‑2 and VEGF/TSP‑1 ratios as potential biomarkers of response in ABCP treated metronomically with Cy+Cel. This finding warrants its confirmation in a higher number of patients.
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Administração Metronômica , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Neoplasias da Mama/sangue , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/patologia , Ciclofosfamida/administração & dosagem , Intervalo Livre de Doença , Feminino , Humanos , Pessoa de Meia-Idade , Pirazóis/administração & dosagem , Ensaios Clínicos Controlados Aleatórios como Assunto , Sulfonamidas/administração & dosagem , Trombospondina 1/sangue , Biomarcadores Tumorais/sangue , Fator A de Crescimento do Endotélio Vascular/sangue , Receptor 2 de Fatores de Crescimento do Endotélio Vascular/sangue , Receptor 3 de Fatores de Crescimento do Endotélio Vascular/sangueRESUMO
Numerous studies have shown that adenosine or adenosine agonists can stimulate angiogenesis. However, the effect of caffeine (a known adenosine receptor antagonist) on angiogenesis has not been previously studied. Accordingly, this study was undertaken to examine the effect of caffeine on angiogenesis and to clarify the mechanism involved. Chick chorioallantoic membrane assays were used to investigate the effect of caffeine on angiogenesis and proliferation assays using human umbilical vein endothelial cells (HUVECs), were used to study its effects on specific aspects of angiogenesis. The expressions of caspase-3 and Bcl-2 were examined by western blotting, immunofluorescence staining was used to identify HUVEC morphological changes, and fluorescence activated cell sorting (FACS) and DAPI staining were used to detect HUVEC apoptosis. Caffeine was found to inhibit blood vessel formation dose-dependently and to inhibit the proliferation of HUVECs time- and dose-dependently. FACS analysis and DAPI staining showed that inhibitory effect of caffeine on HUVEC proliferation was the result of apoptosis and the up-regulation of thrombospondin-1 (TSP-1). Furthermore, TSP-1 levels were down-regulated by NECA but were unaffected by CGS21680, indicating that caffeine regulated TSP-1 expression via adenosine A2B receptor. In addition, caffeine up-regulated caspase-3 and down-regulated Bcl-2 at the protein level. These results suggest that the inhibitory effect of caffeine on angiogenesis is associated, at least in part, with its induction of endothelial cell apoptosis, probably mediated by a caspase-3 dependent mechanism.
Assuntos
Adenosina , Adenosina-5'-(N-etilcarboxamida) , Apoptose , Vasos Sanguíneos , Western Blotting , Cafeína , Caspase 3 , Membrana Corioalantoide , Células Endoteliais , Citometria de Fluxo , Imunofluorescência , Glicosaminoglicanos , Células Endoteliais da Veia Umbilical Humana , Indóis , Fenetilaminas , Receptor A2B de Adenosina , Receptores Purinérgicos P1 , Trombospondina 1 , Regulação para CimaRESUMO
<p><b>OBJECTIVE</b>To investigate the effect of propofol on the expression of thrombospondin-1 (THBS-1) mRNA and protein in purified newborn rat cortical astrocytes in vitro.</p><p><b>METHODS</b>Astrocytes were isolated from newborn rat cortex and grown in culture before exposure to propofol at 3, 10, 30, 100 or 300 µmol/L for 6 h, 12, or 24 h. The mRNA level of THBS-1 was detected by RT-PCR, and the protein level of THBS-1 was detected by immunofluorescence cytochemistry and Western blotting.</p><p><b>RESULTS</b>Propofol exposure caused significantly upregulated THBS-1 level in cultured astrocytes (P<0.05) to a level about 1.3 times higher than that in control cells. The mRNA and protein levels of THBS-1 in cultured rat cortical astrocytes were upregulated by exposures to 10, 30 and 100 µmol/L propofol (P<0.01). High expression of THBS-1 mRNA and protein was detected in the cells with exposures for different durations (P<0.05), especially in the 12 h group (P<0.01).</p><p><b>CONCLUSION</b>Propofol at clinically relevant concentrations can modulate the level of THBS-1 secreted by astrocytes of rat cerebral cortex in vitro.</p>
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Animais , Ratos , Astrócitos , Metabolismo , Células Cultivadas , Córtex Cerebral , Biologia Celular , Propofol , Farmacologia , Ratos Sprague-Dawley , Trombospondina 1 , MetabolismoRESUMO
O Vírus Linfotrópico de células T humanas tipo 1 (HTLV-1) está associado a uma mielopatia (chamada mielopatia associada ao HTLV - HAM/TSP). A trombospondina-1 (TSP-1) é uma proteína da matriz que interfere com a adesão, a motilidade, e a proliferação celular. Níveis deexpressão de RNA mensageiro (mRNA) da trombospondina-1 foram avaliados em indivíduos infectados por HTLV-1: 11 pacientes assintomáticos, 18 com mielopatia ou oligossintomáticos, e 13participantes não-infectados. O RNA de células mononucleares do sangue periférico foi submetido à análise de RT-PCR para trombospondina-1. O número de indivíduos que expressaram esta proteína foi maior no grupo com mielopatia/sintomas (14/18, p igual 0,007). Em geral, a tendência para valores mais elevados de mRNA de trombospondina-1 foi observada no grupo de infectados pelo vírus (p igual 0,062). Os níveis mais elevados de expressão do mRNA foram detectados no início dos sintomas clínicos da HAM/TSP. Estudos adicionais com maior número de amostras são necessários para elucidar melhor o papel desta proteína da matriz na rede inflamatória relacionada à HAM/TSP.
Assuntos
Infecções por HTLV-I , Paraparesia Espástica Tropical/etiologia , Trombospondina 1RESUMO
Angiogenesis is an important process in the pathogenesis of chronic inflammation. We aimed to study the angiogeneic balance in inflammatory bowel disease [IBD] by evaluating the expression of vascular endothelial growth factor [VEGF] and thrombospondin-1 [TSP-1] on colonic epithelial cells, together with the expression of inducible nitric oxide synthase [iNOS]. Twenty-one ulcerative colitis [UC], 14 Crohn's disease [CD], 11 colorectal cancer patients, and 11 healthy controls colonic biopsy samples were evaluated immunohistochemically. The expressions of TSP-1, VEGF, and iNOS in UC and CD groups were higher than expression in healthy control group, all with statistical significance. However, in colorectal cancer group, VEGF and iNOS expressions were increased importantly, but TSP-1 expression was not statistically different from healthy control group's expression. Both TSP-1 and VEGF expressions were correlated with iNOS expression distinctly but did not correlate with each other. Both pro-angiogeneic VEGF and antiangiogeneic TSP-1 expressions were found increased in our IBD groups, but in colorectal cancer group, only VEGF expression was increased. TSP-1 increases in IBD patients as a response to inflammatory condition, but this increase was not enough to suppress pathologic angiogenesis and inflammation in IBD
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Humanos , Masculino , Feminino , Trombospondina 1 , Fator A de Crescimento do Endotélio Vascular , Óxido Nítrico Sintase Tipo II , Colite Ulcerativa , Doença de Crohn , Neoplasias Colorretais , Imuno-HistoquímicaRESUMO
BACKGROUND AND OBJECTIVES: Thrombospondin-1 (TSP-1) is associated with atherosclerosis in animals with diabetes mellitus (DM). But, no study has investigated the role of TSP-1 in human atherosclerosis. This study investigated the relationship among plasma TSP-1 concentration, DM, and coronary artery disease (CAD). SUBJECTS AND METHODS: The study involved 374 consecutive subjects with suspected CAD, who had undergone coronary angiography to evaluate effort angina. Patients were divided into four groups as follows: DM(-) and CAD(-), DM(-) and CAD(+), DM(+) and CAD(-), and DM (+) and CAD(+). RESULTS: We found that plasma TSP-1 levels were higher in patients with DM(+) and CAD(+) (n=103) than those in other patients (n=271) (p<0.01). A multivariate analysis showed that male gender {odds ratio (OR), 2.728; 95% confidence interval (CI), 1.035-7.187}, high density lipoprotein-cholesterol (OR, 0.925; 95% CI, 0.874-0.980), glycated hemoglobin (OR, 1.373; 95% CI, 1.037-1.817), and plasma TSP-1 (OR, 1.004; 95% CI, 1.000-1.008) levels were independently associated with the presence of CAD in patients with DM. CONCLUSION: Plasma TSP-1 levels were higher in patients with DM(+) and CAD(+) than those in other patients, and plasma TSP-1 levels were independently associated with the presence of CAD in patients with DM. These findings show a possible link between human plasma TSP-1 concentration and CAD in patients with DM.
Assuntos
Animais , Humanos , Masculino , Hidróxido de Alumínio , Aterosclerose , Carbonatos , Angiografia Coronária , Doença da Artéria Coronariana , Vasos Coronários , Diabetes Mellitus , Hemoglobinas , Análise Multivariada , Plasma , Trombospondina 1RESUMO
Breast cancer is the most common cancer in women worldwide. It is necessary to identify biomarkers for early detection, to make accurate prognoses, and to monitor for any recurrence of the cancer. In order to identify potential breast cancer biomarkers, we analyzed the plasma samples of women diagnosed with breast cancer and age-matched normal healthy women by mTRAQ-based stable isotope-labeling mass spectrometry. We identified and quantified 204 proteins including thrombospondin-1 (THBS1) and bromodomain and WD repeat-containing protein 3 (BRWD3) which were increased by more than 5-fold in breast cancer plasma. The plasma levels of the two proteins were evaluated by Western blot assay to confirm for their diagnostic value as serum markers. A 1.8-fold increase in BRWD3 was observed while comparing the plasma levels of breast cancer patients (n = 54) with age-matched normal healthy controls (n = 30), and the area under the receiver operating characteristic curve (AUC) was 0.917. THBS1 was detected in pooled breast cancer plasma at the ratio similar to mTRAQ ratio (> 5-fold). The AUC value for THBS1 was 0.875. The increase of THBS1 was more prominent in estrogen receptor negative and progesterone receptor negative patients than receptor-positive patients. Our results are evidence of the diagnostic value of THBS1 in detecting breast cancer. Based on our findings, we suggest a proteomic method for protein identification and quantification lead to effective biomarker discovery.
Assuntos
Adulto , Feminino , Humanos , Pessoa de Meia-Idade , Neoplasias da Mama/diagnóstico , Detecção Precoce de Câncer , Perfilação da Expressão Gênica , Patologia Molecular/métodos , Valor Preditivo dos Testes , Prognóstico , Proteômica , Trombospondina 1/sangue , Fatores de Transcrição/sangue , Biomarcadores Tumorais/sangueRESUMO
<p><b>OBJECTIVE</b>Pancreatic cancer is one of the most deadly cancers, which is characterized by its high metastatic potential. S100A4 is a major prometastatic protein involved in tumor invasion and metastasis which precise role in pancreatic cancer has not been fully investigated. We knocked down the S100A4 gene in the Bxpc-3 pancreatic cancer cell line via RNA interference to study the changes in cell behavior.</p><p><b>METHODS</b>Real-time polymerase chain reaction and western blotting were used to detect mRNA and protein expression levels of S100A4, matrix metalloproteinase (MMP)-2, E-cadherin and thrombospondin (TSP)-1. Transwell chambers were used to detect the migration and invasion abilities; a cell adhesion assay was used to detect adhesion ability; colony forming efficiency was used to detect cell proliferation; flow cytometry was used to detect apoptosis.</p><p><b>RESULTS</b>S100A4 mRNA expression was reduced to 17% after transfection with S100A4-siRNA, and protein expression had a similar trend. mRNA and protein expression of MMP-2 was reduced and that of E-cadherin and TSP-1 was elevated, indicating that S100A4 affects their expression. S100A4-silenced cells exhibited a marked decrease in migration and invasiveness and increased adhesion, whereas overall proliferation and apoptosis were not overtly altered.</p><p><b>CONCLUSION</b>S100A4 and its downstream factors play important roles in pancreatic cancer invasion, and silencing A100A4 can significantly contain the invasiveness of pancreatic cancer.</p>