High PAI-1 Research Articles

Concentrations of plasminogen activator inhibitor type1 (PAI-I) and urokinase-type plasminogen aktivator (uPA) in primary breast cancer (BC) tissue and corresponding axillary lymph node metastasis.

Abstract Abstract #2046 Background: Components of the plasminogen activator system play a key role in tumor invasion and metastasis. Tumor tissue levels of urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 showed a strong prognostic impact in node-negative and node-positive primary BC. uPA/PAI-1 were the first novel prognostic factors in primary BC reaching the highest level of evidence for their clinical utility and they have recently been recommended by ASCO for clinical decision making.
 Material and Methods: For the first time, correlations between uPA and PAI-1 concentrations in 73 primary tumor tissues and corresponding axillary lymphnodes (ALN) in node-negative (pN0) and node-positive (pN+) BC (1991-98) and their prognostic impact on disease-free (DFS) and overall survival (OS) were evaluated.
 Results: Median follow-up was 56 months. Overall, PAI-I (p=0,014) and uPA concentrations (p<0,001) were significantly higher in tumour tissue than in the corresponding ALN. There was no significant difference of PAI-1 in primary tumor tissue between pN0 and pN+ disease. In involved ALN (pN+), concentrations of PAI-1 but not of uPA, were significantly higher than in pN0 (p<0.001). Moreover, patients with higher PAI-1 concentrations in the corresponding ALN than in their primary tumor had a significantly shorter DFS (p<0,001) and OS (p=0.005).
 Discussion: Our clinical findings support the basic research data implicating uPA and PAI-1 as key proteolytic factors in metastasis-asociated processes such as adhesion, angiogenesis, invasion, proliferation and migration. For the first time, we are able to demonstrate that particularly PAI-1 seems to be involved in tumor metastasis in primary breast cancer with its concentrations being higher in involved ALN than in the primary tumor. Besides uPA/PAI-1 being validated and clinically useful prognostic factors, interference with the uPA/PAI-1 system thus promises to be an interesting therapeutic concept in BC. Citation Information: Cancer Res 2009;69(2 Suppl):Abstract nr 2046.

The leukemogenic activity of TaxHTLV-1 during human alphabeta T cell development

The regulatory Tax protein of HTLV-1 (Human T-cell Leukaemia Virus type 1) is critically involved in the initiation of ATL (adult T-cell leukaemia). Indeed, Tax provides infected T-cells with a growth advantage and with the potential to get transformed through the deregulation of cell-cycle progression and the acquisition of genetic alterations. Considering that leukemias are induced by disturbances in hematopoietic cells development, we hypothesize that the expression of Tax in human immature thymocytes is a prerequisite to the emergence of ATL cells. Studies of alph abeta T-cell development in the thymus have shown that beta-selection, an early important checkpoint, is regulated by transcription factors that are decisive in the control of cell proliferation, differentiation and survival. Interestingly, Tax is endowed with the ability to interfere with the activity of these transcription factors. We therefore propose that the HTLV-1 infection of these specific target thymocytes leads to a transcriptional deregulation of early alphabeta T cell development, thus inducing a pre-leukemogenic event that favours the subsequent proliferation of ATL cells.

Analysis of NM23 Protein and Components of Plasminogen Activation System in Tumors of Patients with Stomach Cancer with Consideration for Disease Clinical Picture and Morphology

Immunohistochemical analysis of the expression and distribution of nm23 protein and biochemical analysis of the main components of plasminogen activation system in tumors were carried out in stomach cancer patients. The data indicate that the expression of nm23 protein in malignant epithelial gastric tumors is heterogeneous, characterized by cytoplasmic and nuclear immunoreactivity. Reduced expression of the marker is more typical of poorly or undifferentiated tumors. The expression of nm23 protein positively correlated with tPA level (r(s)=0.4; p=0.01) and did not correlate with the content of uPA and PAI-1 in tumors. High PAI-1 values in tumors (>0.5 ng/mg protein) significantly correlated with lower 3-year overall survival of stomach cancer patients. These data confirm the role of the studied proteins in invasion and metastases of malignant tumors and suggest a relationship between changes in the expression of nm23 protein and mechanisms of stomach cancer progress.

Abstract 5105: The Relationship of Inflammation and Thrombosis with Right Ventricular Mass and Function: The MESA-Right Ventricle Study

Right ventricular (RV) mass reflects response to afterload, whereas RV ejection fraction (RVEF) is an indicator of RV function. Pulmonary vascular disease is characterized by hemostatic abnormalities and inflammation and may affect RV mass and function. We hypothesized that increased levels of inflammatory markers (ICAM-1, TNF-α, E-Selectin, MMP-3, and MMP-9) would be associated with higher RV mass and lower RVEF, and increased levels of hemostatic markers (tissue factor, PAI-1, and CD40L) and decreased levels of TFPI would be associated with higher RV mass and lower RVEF. The Multi-Ethnic Study of Atherosclerosis (MESA) performed interpretable cardiac MRIs on 5,004 participants without clinical cardiovascular disease at six field centers. 785 randomly selected patients had plasma biomarkers measured, 730 of whom had complete interpretation of RV measures. Endocardial margins of the RV were manually contoured on diastolic and systolic images. End-diastolic volume (EDV) and end-systolic volume (ESV) were calculated by using a summation of disks method (“Simpson’s Rule”). RVEF was calculated from (EDV-ESV)/EDV. RV mass was determined from cine images of the heart. Biomarkers were analyzed in quintiles using multivariate linear regression. The 730 participants in the study sample were 59 ± 10 (mean ± SD) years old, 58% female, 45% Caucasian, 21% African-American, 11% Chinese-American, and 23% Hispanic. The RV mass was 22 g ± 5 g and RVEF was 69% ± 7%. Higher PAI-1 levels were associated with lower RV mass, after adjustment for age, gender, race, height, and weight (test for trend, p = 0.036, n = 710). Higher MMP-9 levels were associated with lower RV mass (test for trend, p = 0.006). Neither hemostatic nor inflammatory biomarkers were associated with RVEF. RV mass was inversely associated with PAI-1, a serine protease which inhibits thrombolysis, and MMP-9, which breaks down the extracellular matrix. The mechanism of these determinants of RV structure and function should be investigated.

Tumor marker utility and prognostic relevance of cathepsin B, cathepsin L, urokinase-type plasminogen activator, plasminogen activator inhibitor type-1, CEA and CA 19-9 in colorectal cancer

BackgroundCathepsin B and L (CATB, CATL), urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 play an important role in colorectal cancer invasion. The tumor marker utility and prognostic relevance of these proteases have not been evaluated in the same experimental setting and compared with that of CEA and CA-19-9.MethodsProtease, CEA and CA 19-9 serum or plasma levels were determined in 56 patients with colorectal cancer, 25 patients with ulcerative colitis, 26 patients with colorectal adenomas and 35 tumor-free control patients. Protease, CEA, CA 19-9 levels have been determined by ELISA and electrochemiluminescence immunoassay, respectively; their sensitivity, specificity, diagnostic accuracy have been calculated and correlated with clinicopathological staging.ResultsThe protease antigen levels were significantly higher in colorectal cancer compared with other groups. Sensitivity of PAI-1 (94%), CATB (82%), uPA (69%), CATL (41%) were higher than those of CEA or CA 19-9 (30% and 18%, respectively). PAI-1, CATB and uPA demonstrated a better accuracy than CEA or CA 19-9. A combination of PAI-1 with CATB or uPA exhibited the highest sensitivity value (98%). High CATB, PAI-1, CEA and CA 19-9 levels correlated with advanced Dukes stages. CATB (P = 0.0004), CATL (P = 0.02), PAI-1 (P = 0.01) and CA 19-9 (P = 0.004) had a significant prognostic impact. PAI-1 (P = 0.001), CATB (P = 0.04) and CA 19-9 (P = 0.02) proved as independent prognostic variables.ConclusionAt the time of clinical detection proteases are more sensitive indicators for colorectal cancer than the commonly used tumor markers. Determinations of CATB, CATL and PAI-1 have a major prognostic impact in patients with colorectal cancer.

Coagulation and inflammation markers during atypical or typical antipsychotic treatment in schizophrenia patients and drug-free first-degree relatives

BackgroundClinical studies suggest that the second generation antipsychotics (APs) clozapine and olanzapine and to a lesser extent the typical antipsychotics may be associated with a procoagulant and proinflammatory state that promotes venous thromboembolism. We evaluated here several blood factors associated with coagulation and inflammation in AP-treated schizophrenia patients and their first-degree relatives. MethodsProcoagulant factors (fibrinogen and plasminogen activator inhibitor [PAI-1]), the anticoagulant factor antithrombin III [AT-III], and inflammation-related factors (C-reactive protein [CRP] and leptin) were assessed in patients chronically treated with clozapine (n=29), olanzapine (n=29), typical APs (n=30) and first degree relatives of clozapine (n=23) and olanzapine subjects (n=11). ResultsThe typical AP group had the highest CRP level (p=0.013) in spite of having the lowest body mass index (BMI). Patients as a single group had higher CRP levels than relatives (p=0.003). The typical AP group also had the highest AT-III levels (p=0.021). Fibrinogen levels did not differ between the groups (p=0.13). Olanzapine patients displayed the highest PAI-1 and leptin levels among the drug-treated subjects, but values were similar to those observed in their relatives, and were significantly correlated with the BMI. ConclusionsA homogeneous negative profile of high inflammation and procoagulant factors along with low levels of anticoagulants was not detected in any group. While preliminary, our results suggest that the observed abnormalities were not related to a direct drug effect, but to elevated BMI (high PAI-1 and leptin in olanzapine-treated patients). We speculate that the high CRP in the typical AP group might be related to poor lifestyle habits, but this must we confirmed in future studies.

A comparison among HER2, TP53, PAI-1, angiogenesis, and proliferation activity as prognostic variables in tumours from 408 patients diagnosed with early breast cancer

Background. The prognostic potential of HER2, TP53 mutations, PAI-1 protein levels, angiogenesis and proliferation were investigated in tumours from 408 patients with early breast cancer followed >10 years. One hundred and sixty seven patients (41%) died from breast cancer. Materials and methods. Tumour sections were stained for HER2, CD34, and MIB-1. HER2 scores were based on staining intensity, 3+ being considered HER2+. Angiogenesis was scored by the Chalkley method. MIB-1 was evaluated using systematic random sampling. PAI-1 was measured by ELISA. TP53 mutations were evaluated by DGGE analysis and DNA sequencing. Results. Ninety one patients (22%) were HER2 positive. TP53 was mutated in 101 cases (25%). Median PAI-1, Chalkley and MIB-1 was 0.72 ng/mg protein (range, 0–90 ng/mg protein), 5.00 (range, 2.67–12.00) and 15% (range, 1–83%). MIB-1 was correlated with HER2+, Chalkley counts, TP53 mutations (all p <0.0001), and PAI-1 (p =0.002). In univariate analyses with DSS as endpoint, HER2+ (p <0.0001), mutated TP53 (p <0.0001), high Chalkley (p =0.008), MIB-1 (p =0.002), tumour size (p =0.008), grade (p <0.0001), negative estrogen receptor (p =0.0001), and lymph node status (p <0.0001) were prognostic markers. Among node-negative patients, HER2+ (p =0.0002), mutated TP53 (p =0.001), high PAI-1 levels (p =0.02), and grade (p =0.03) indicated poor DSS. In node-positive patients, HER2+ (p =0.0002), mutated TP53 (p <0.0001), MIB-1 (p =0.01), Chalkley scores (p =0.007), negative estrogen receptor (p <0.0001) and grade (p =0.001) indicated poor prognosis. In multivariate analysis, metastatic nodes (1–3 positive: RR 1.56 95% CI 1.02–2.38; >3 positive: RR 3.70 95% CI 2.54–5.38), HER2+ (RR 1.91, 95% CI 1.35–2.70), mutated TP53 (RR 1.70, 95% CI 1.21–2.38), PAI-1 (RR 1.04, 95% CI 1.01–1.07) and grade 3 (RR 1.96, 95% CI 1.83–3.22) were independent markers of poor outcome. Conclusion. Compared to PAI-1 protein levels, Chalkley counts and MIB-1, HER2+ and mutations of TP53 were the strongest independent markers of poor prognosis irrespective of nodal status.

Association Between Polysomnographic Measures of Disrupted Sleep and Prothrombotic Factors

Background:Subjective sleep disturbances have been associated with increased risk of coronary artery disease (CAD). We hypothesized that disrupted sleep as verified by polysomnography is associated with increased levels of prothrombotic hemostasis factors previously shown to predict CAD risk.Methods:Full-night polysomnography was performed in 135 unmedicated men and women (mean age ± SD, 36.8 ± 7.8 years) without a history of sleep disorders. Morning fasting plasma levels of von Willebrand Factor (VWF) antigen, soluble tissue factor (sTF) antigen, d-dimer, and plasminogen activator inhibitor (PAI)-1 antigen were determined. Statistical analyses were adjusted for age, gender, ethnicity, body mass index, BP, and smoking history.Results:Higher total arousal index (ArI) was associated with higher levels of VWF (β = 0.25, p = 0.011, ΔR2= 0.045), and longer wake after sleep onset was associated with higher levels of sTF (β = 0.23, p = 0.023, ΔR2= 0.038). More nighttime spent at mean oxygen saturation < 90% (β = 0.20, p = 0.020, ΔR2= 0.029) and higher apnea-hypopnea index (AHI) [β = 0.19, p = 0.034, ΔR2= 0.024] were associated with higher PAI-1. There was a trend for a relationship between mean oxygen desaturation < 90% and PAI-1 (p = 0.053), even after controlling for AHI. Total ArI (β = 0.28, p = 0.005, ΔR2= 0.056) and WASO (β = 0.25, p = 0.017, ΔR2= 0.042) continued to predict VWF and sTF, respectively, even after controlling for AHI.Conclusions:Polysomnographically verified sleep disruptions were associated with prothrombotic changes. Measures of sleep fragmentation and sleep efficiency were related to VWF and sTF, respectively. Apnea-related measures were related to PAI-1. Our findings suggest that sleep disruptions, even in a relatively healthy population, are associated with potential markers of prothrombotic cardiovascular risk.

Plasminogen activator inhibitor (PAI)-1 in vascular inflammation and thrombosis

Impaired fibrinolysis may be associated with development of atherothrombotic cardiovascular disease (CVD) in metabolic syndrome or type 2 diabetes. Plasma plasminogen activator inhibitor (PAI)-1, a potent inhibitor of fibrinolysis, is elevated in a number of clinical situations that are associated with high incidence of CVD. Impaired fibrinolysis resulting from high plasma PAI-1 can lead to excessive fibrin accumulation within vessels, resulting in atherothrombosis. Increased expression of PAI-1 is found in atherosclerotic lesions in humans, especially atherosclerotic plaques in patients with type 2 diabetes. This increased vascular expression of PAI-1 promotes neointima formation via accumulation of fibrin or fibrinogen as a result of inhibited clearance of platelet-fibrin thrombi. PAI-1, an acute phase protein, also could be involved in vascular inflammation. PAI-1 may be associated not only systemically but also locally with development of CVD.

Prognostic Value of PAI1 in Invasive Breast Cancer: Evidence That Tumor-Specific Factors Are More Important Than Genetic Variation in Regulating PAI1 Expression

Plasminogen activator inhibitor-1 (PAI1) can promote cancer progression, and its protein expression in tumors is an independent indicator of poor prognosis in many forms of cancer. Here, we show that high PAI1 mRNA levels also predict for shorter overall survival in two independent breast cancer data sets, highlighting the importance of its transcriptional regulation. The -675insG (4G/5G) single-nucleotide polymorphism in the PAI1 gene promoter has been shown to influence PAI1 transcription, with the 4G allele eliciting higher reporter gene expression in vitro and higher levels of circulating PAI1 in vivo. Nevertheless, its genotypic distribution in 2,539 British women with invasive breast cancer was virtually identical to that seen in 1,832 matched controls (P = 0.72), and annual mortality rates for 4G4G, 4G5G, and 5G5G cases were 2.6%, 2.8%, and 3.1% per year, respectively (P = 0.10). Thus, there was no association with breast cancer incidence or outcome, and in a separate set of breast cancers, the 4G/5G single-nucleotide polymorphism showed no association with PAI1 mRNA expression (P = 0.85). By contrast, connective tissue growth factor (CTGF), which can regulate PAI1 expression in culture, was associated with PAI1 expression in three independent cohorts (P << 0.0001). In addition, PAI1 gene copy number differences in the tumors were correlated with PAI1 mRNA expression (P = 0.0005) and seemed to affect expression independently of CTGF. Thus, local factors, such as CTGF and genomic amplification, seem to be more important than germ line genetic variation in influencing PAI1 expression and its untoward effects in breast cancer.

Urokinase-type plasminogen activator and its inhibitor in thyroid neoplasms: a cytosol study

Higher levels of urokinase-type plasminogen activator (uPA) and its inhibitor (PAI-1) are linked to the poor prognosis in a variety of malignances. uPA and PAI-1 were expressed in most thyroid carcinomas, as had been measured immunohistochemically. However, no relationship between their expression and clinicopathological parameters were found. Aim of the present study was to investigate the expression and clinical relevance of uPA and PAI-1 in thyroid cancer. uPA and PAI-1 in paired cytosol samples of thyroid tumor and normal tissue were determined in 23 patients using enzyme-linked immunosorbent assay and correlated to the known prognostic features. Both uPA and PAI-1 concentrations were significantly higher in malignant thyroid tumors (uPA=1.342 +/- 2.944 and PAI-1=17.615 +/- 31.933 ng/mg protein) than in normal tissue (uPA=0.002 +/- 0.009, P=0.011 and PAI-1=2.333 +/- 0.338 ng/mg protein, P=0.001) with positive correlation of the two proteins in the tumors. There were no differences in proteins' levels between benign tumors and normal tissue. Both proteins' concentrations were significantly different among various histological grades (uPA P=0.024 and PAI-1 P=0.017), showing higher values in higher tumor grades (grade I uPA=0.116 +/- 0.247 and PAI-1=4.802 +/- 4.151 ng/mg protein; grade III uPA=8.45 +/- 2.192 and PAI-1=94.65 +/- 59.468 ng/mg protein). The uPA and PAI-1 levels showed significant differences among different histological types of thyroid cancer (uPA P=0.049 and PAI-1=0.017). The lowest values were in adenomas (uPA=0.013 +/- 0.025 and PAI-1=2.785 +/- 1.069 ng/mg protein) and the highest in anaplastic carcinomas (uPA=8.45 +/- 2.192 and PAI-1=94.65 +/- 59.468 ng/mg protein). uPA and PAI-1 were significantly higher in anaplastic vs. well-differentiated cancers (uPA P=0.014 and PAI-1 P=0.026), if extrathyroidal invasion (uPA P=0.019 and PAI-1 P=0.009) or distant metastases (uPA P=0.006 and PAI-1 P=0.003) had been present, and in tumors whose size exceeded 1 cm in diameter (uPA P=0.009 and PAI-1 P=0.035). Only PAI-1, but not uPA was significantly higher in multicentric vs. solitary tumors (P=0.012) and lymph node positive compared to lymph node negative patients (P=0.042). The differences of uPA and PAI-1 did not reach the significant level when patients with well-differentiated tumors below and above 40 years of age had been compared. Survival analysis revealed the significant impact of both uPA and PAI-1 on the Progression-Free Survival (PFS) (38.84 vs. 3.67 months for patients with low and high uPA, respectively, P<0.001; 38.2 vs. 12 months for patients with low and high PAI-1, respectively, P=0.016). The correlation of high uPA and PAI-1 with the known prognostic factors of poorer outcome and with lower PFS rate in patients with thyroid cancers proved that these proteins could be an additional prognostic parameter.

ID: 311 High PAI-1 and the malignant phenotype: exogenous PAI-1 signals to tumors cells via uPA/uPAR/LDLR-family system and ERK1/2 pathway inducing increased adhesion and integrin redistribution

ID: 311 High PAI-1 and the malignant phenotype: exogenous PAI-1 signals to tumors cells via uPA/uPAR/LDLR-family system and ERK1/2 pathway inducing increased adhesion and integrin redistribution

Molecular risk estimation and adjuvant chemotherapy in node-negative breast cancer patients—A status report of the prospective clinical trial NNBC 3-Europe

10503 Background: Feasibility of risk assessment in node negative breast cancer using the invasion markers urokinase-type plasminogen activator uPA and its inhibitor PAI-1 has been demonstrated in several prospective and retrospective studies and in a large meta analysis. Patients with low uPA and/or PAI-1 concentrations in tumor tissue levels have an excellent 5-year overall survival (&gt;95%) even without any adjuvant therapy. Use of these molecular markers may spare adjuvant chemotherapy in approximately one half of node-negative breast cancer patients. Particulary, patients with an intermediate grade tumor can easily be differentiated in low and high risk. In addition, patients with high uPA and/or PAI-1 level seem to benefit from adjuvant chemotherapy. The NNBC-3-Europe trial seeks to answer two questions: 1. Is risk assessment by molecular markers uPA/PAI-1 superior to that by clinico-pathological factors regarding identification of low-risk patients? 2. Is adjuvant chemotherapy with anthracycline-taxane sequence (FEC-Docetaxel) superior to standard FEC in high-risk patients? Methods: In the NNBC-3-Europe trial, participating centres opt to either perform risk estimation by clinico-pathological factors or by the uPA and PAI-1 concentration in tumor tissue biopsies. Low-risk patients will be observed without adjuvant chemotherapy. High-risk patients are randomised based on the kind of adjuvant chemotherapy. Patients with steroid hormone receptor positive tumors receive adequate endocrine therapy. Results: Of the first 506 patients in the study arm with molecular risk assessment, 62 had a grade 1 tumors, and 184 had grade 3 tumors. Among grade 2 tumors (n = 260), 87 had low uPA/PAI-1. 173 patients presented with high levels. About 30% of the patients were allocated to the low-risk group using uPA/PAI-1. Conclusions: An adjuvant chemotherapy trial based on uPA/PAI-1 determination in the primary tumor is feasible in a multicentre setting. Applying these molecular markers for risk assessment, at this stage of the study, adjuvant chemotherapy could be spared in almost one third of N0 patients. It is performed in association with the EORTC PathoBiology Group Receptor und the German AGO Breast Group. [Table: see text]

The effect of high plasma levels of angiotensin-converting enzyme (ACE) and plasminogen activator inhibitor (PAI-1) on the reperfusion after thrombolytic therapy in patients presented with acute myocardial infarction

The resistance to thrombolytic agents and delays in reperfusion occur in more than 30% after acute myocardial infarction. This may play an important role in the unsuccessful recanalization after thrombolytic therapy. The aim of this study is to assess the clinical and biochemical markers of reperfusion after different types of thrombolytic therapy and to find out the relationship between PAI-1 and ACE serum levels and the short-term outcome. Pretreatment ACE and PAI-1 plasma levels of 184 patients with acute myocardial infarction, treated with thrombolytic therapy were determined. Failure of thrombolysis was considered when reperfusion was delayed as assessed by noninvasive reperfusion criteria, reinfarction, and impaired left ventricular function. High plasma level of ACE (> 50 U/L), PAI-1 (> 43 ng/ml) and both was found in 57, 108 and 32 patients respectively. Subjects with high ACE plasma levels were characterized by impaired LV systolic function (79.0% vs. 75.0%), new Q-wave (88.4% vs. 74.2%), less reperfusion arrhythmia (19.3% vs. 22.8%) and prolonged hospitalization (70% vs. 66%) but no statistical significance was observed. High enzymes levels of PAI-1 were observed with higher incidence of anterior myocardial infarction (50.0% vs. 41.0%), lesser ST segment resolution (65.6% vs. 58.8%), reinfarction (6.3% vs. 5.9%), and impaired LV systolic function (90.6% vs. 76.0%), and prolonged hospitalization (70.4% vs. 63.4). There was a statistically significant difference between thrombolytic agents in the presence of high ACE regarding hospital overstay (p = 0.02). While the presence of high PAI-1 was significantly affect the degree of ST-segment resolution (p = 0.03). High plasma ACE and/or PAI-1 plays a considerable role in the higher incidence of unsuccessful reperfusion and impaired left ventricular function after thrombolytic therapy. A rapid diagnostic tool that enables physician of detecting those enzymes before giving thrombolytic therapy may change the strategy of treatment to offer another effective revascularization method.

The 4G/4G PAI‐1 genotype is associated with elevated plasma PAI‐1 levels regardless of variables of the metabolic syndrome and smoking status. A population‐based study in Spanish population

Reported data about the effect of the 4G/5G PAI-1 polymorphism on plasma PAI-1 levels are controversial. This study was designed to determine the relative effect of the 4G/5G PAI-1 polymorphism on high plasma PAI-1 levels after adjustment for metabolic syndrome - related variables, and to test if this effect is modified by the smoking status. Six hundred and thirty one unrelated subjects (292 men; 35-74 years), from a cross-sectional population-based epidemiological survey in the province of Segovia (Spain) were studied. The higher frequency of high PAI-1 levels was found in 4G/4G subjects (5G/5G 19.4%, 4G/5G 21.6%, 4G/4G 33.7%, p = 0.003). A multiple regression model, adjusted for gender, age, BMI, waist circumference, triglycerides, HDL-cholesterol, HOMA IR and leptin, showed this adjOR: 4G/4G vs 5G/5G: 2.22, p = 0.008. When smoking status - 4G/5G PAI-1 interaction was included as an independent variable these results were not modified. Our results indicate that the 4G/4G PAI-1 genotype might be strongly associated with high PAI-1 levels regardless of metabolic syndrome-related variables and smoking status.

Plasminogen activator inhibitor activity, 4G5G polymorphism of the plasminogen activator inhibitor 1 gene, and first-trimester miscarriage in women with polycystic ovary syndrome

We assessed whether hypofibrinolytic plasminogen activator inhibitor 1 (PAI-1 activity) showed an independent association with first-trimester miscarriage in the 430 women with polycystic ovary syndrome (PCOS) who had previous pregnancies (from a cohort of 967 women with PCOS). Prospectively, we hypothesized that Glucophage (Bristol-Myers Squibb, Princeton, NJ) promotes successful live births in women with PCOS by lowering PAI-1 activity before conception and maintaining further reductions of PAI-1 activity during the first trimester of pregnancy. We also assessed whether PAI-1 activity levels were independently related to PAI-1 genotype and to modifiable risk factors body mass index (BMI), insulin, and triglyceride. By stepwise logistic regression, with the dependent variable being previous pregnancy outcomes at 3 levels (live birth pregnancies only [n = 208]; both ≥1 live birth and ≥1 first-trimester miscarriage [n = 111]; or first-trimester miscarriages only [n = 71]) and explanatory variables PAI-1 genotype, PAI-1 activity, insulin, homeostasis model assessment of insulin resistance, BMI, and triglyceride, PAI-1 activity was positively associated with first-trimester miscarriage ( P = .004). For each 5 IU/mL increment in PAI-1 activity, the risk being in an adverse first-trimester miscarriage category increased (odds ratio, 1.12; 95% confidence interval, 1.04-1.20). Prospectively, from pretreatment to the last preconception visit on Glucophage, in 30 women who subsequently had live births, PAI-1 activity fell 44%, but rose 19% in 23 women with first-trimester miscarriage ( P = .03). In the 30 women with live birth pregnancies, median PAI-1 activity fell continuously from pretreatment through the first trimester (from 16.8 to 6.7 IU/mL), whereas PAI-1 activity was either unchanged or rose in women with first-trimester miscarriage. Of the 921 women with PCOS who had 4G5G data, 718 (78%) had 4G4G-4G5G genotypes vs 87 (69%) of 126 normal female controls ( χ 2 = 4.95, P = .026). The 4G allele frequency was 53% in women with PCOS vs 46% in controls ( χ 2 = 4.3, P = .04). Of the 866 women with PCOS who had PAI-1 activity data, by stepwise regression, positive independent determinants of PAI-1 activity included BMI (partial R 2 = 10.6%, P < .0001), insulin (partial R 2 = 2.8%, P < .0001), triglyceride (partial R 2 = 1.1%, P = .0009), and the 4G4G-4G5G genotype (partial R 2 = 1%, P = .0011). The PAI-1 gene 4G polymorphism is more common in women with PCOS than in normal women and, in concert with obesity, hyperinsulinemia, and hypertriglyceridemia, contributes to treatable, hypofibrinolytic, miscarriage-promoting, high PAI-1 activity. Preconception and first-trimester decrements in PAI-1 activity on Glucophage are associated with live births, whereas increments or no change in PAI-1 activity despite Glucophage appears to be associated with first-trimester miscarriage.

The Inhibitory Effect of siRNAs on The High Glucose-Induced Overexpression of TGF-β1 in Mesangial Cells

Diabetic nephropathy is characterized by an expansion of the glomerular mesangium, caused by mesangial cell proliferation and an excessive accumulation of extracellar matrix (ECM) proteins, which eventually leading to glomerulosclerosis. TGF-β1 was found to play an important role in the accumulation of ECM in the kidney. In this study, TGF-β1 RNA interference was used as an effective therapeutic strategy. The inhibitory effect of TGF-β1 small interfering RNAs (siRNAs) on the high glucose-induced overexpression of TGF-β1 in rat mesangial ceys (RMCs). A high levels of glucose induces TGF-β1 mRNA and protein, and TGF-β1 siRNAs reduce the ability of high glucose to stimulate their expression. We also examined the inhibitory effect of TGF-β1 siRNAs on the expression of plasminogen activator inhibitor (PAI)-1 and Collagen Type I which are down-regulators of TGF-β1. The expression of TGF-β1, PAI-1 and Collagen Type I was increased in RMCs that were stimulated by 30 mM glucose. TGF-β1 siRNAs reduces high glucose-induced TGF-β1, PAI-1, and Collagen Type I mRNA and protein expression in a dose-dependent manner. In conclusion, the present study demonstrates that TGF-β1 siRNAs effectively inhibits TGF-β1 mRNA and protein expression in RMCs. These suggest that TGF-β1 siRNAs through RNAi may be a useful tool for developing new therapeutic applications for the treatment of diabetic nephropathy.

The complex between urokinase (uPA) and its type-1 inhibitor (PAI-1) in pulmonary adenocarcinoma: Relation to prognosis

In a lung cancer population comprising tumor tissue from 99 pulmonary adenocarcinoma patients, the relationship between tumor tissue level of the complex formed of urokinase (uPA) and its type-1 inhibitor (PAI-1) and survival was studied. The study included patient material previously investigated for the prognostic impact of PAI-1 on survival. Standard clinical parameters were available and the patients had a median survival time of 25 months. An ELISA established to measure preformed uPA-PAI-1 complexes was applied to the tumor extracts and previously measured data on uPA and PAI-1 levels were available. The amounts of uPA-PAI-1 complex measured in pulmonary adenocarcinoma tissue were within the same range as previously reported in breast cancer tissue (0.11-5.74 ng/mg protein). uPA and PAI-1 levels were weakly correlated to the uPA-PAI-1 complex, r = 0.52 and r = 0.47, respectively, and no relation was found between uPA-PAI-1 complex and any of the clinical parameters. However, a significant prognostic impact of PAI-1 on prognosis was demonstrated (HR = 1.62, p = 0.04). Patients with high PAI-1 and low uPA-PAI-1 complex were found to have a significantly poorer survival than patients with low PAI-1 and high uPA-PAI-1 complex (HR = 3.06, p = 0.01). This is the first investigation of the prognostic impact of uPA-PAI-1 complex in a tumor type other than breast cancer, showing low levels of uPA-PAI-1 complex in combination with high levels of PAI-1 to be associated with poor prognosis. To understand these interactions and the clinical importance of the tissue levels of uPA, PAI-1 and uPA-PAI-1 complex, the results suggest further exploratory studies of the components in pulmonary adenocarcinomas and other cancers.

Deflazacort modulates the fibrinolytic pattern and reduces uPA-dependent chemioinvasion and proliferation in rheumatoid arthritis synoviocytes

Extracellular fibrinolysis, controlled by the cell-associated fibrinolytic system (urokinase plasminogen activator, uPA; uPA receptor, uPAR; plasminogen activator inhibitor type-1, PAI-1), is involved in cartilage damage generation and in rheumatoid arthritis (RA) synovitis. Since steroids reduce the rate of radiological progression of RA, we planned to evaluate in healthy and RA synoviocytes the effects of the steroid deflazacort on uPA, uPAR and PAI-1 expression, and subsequent phenotypic modifications in terms of uPA/uPAR-dependent invasion and proliferation. uPA, uPAR and PAI-1 levels were studied by ELISA, RT-PCR (uPAR) and zymography (uPA) in synoviocytes from four RA patients and four healthy controls. Chemoinvasion was assessed by the Boyden chamber invasion assay, using Matrigel as the invasion substrate. Proliferation was evaluated by cell counting. Both invasion and proliferation were measured upon treatment with deflazacort 5 muM with or without parallel stimulation with uPA 500 ng/ml or in the presence of monoclonal anti-uPA and anti-uPAR antibodies. Invasion and proliferation of RA synoviocytes require a proper functional balance of the fibrinolytic system. Both deflazacort and monoclonal antibodies against uPA and uPAR reduced expression and activity of the system, thus inhibiting invasion and proliferation. In RA synoviocytes, deflazacort induced higher PAI-1 and lower uPA and uPAR levels, as well as a decrease in uPA enzymatic activity. The levels of uPAR mRNA were concomitantly reduced, as was uPA-induced chemoinvasion. All these effects were also shown in controls, though to a lesser extent. Deflazacort might control RA synovial proliferation and invasion by differential modulation of single members of the fibrinolytic system.

Fetal scarless healing: The role of TGF-beta 3 and pro-fibrotic molecule PAI-1

Abstract Introduction: Early mammalian fetal wounds heal skin wounds scarlessly, by regeneration rather than repair, through unknown mechanisms. During normal wound healing, plasmin plays a vital role in matrix remodeling, a critical determinant of scar formation. Its activity depends on the ratio of plasminogen activator inhibitor-1 (PAI-1, a marker of fibrosis) and urokinase-type plasminogen activator (uPA), which we have found to increase during fetal development. TGF-beta is essential in tissue injury repair. Of the three mammalian isoforms, TGF-beta 3 appears to be anti-scarring. Whilst TGF-beta 1 is a potent inducer of PAI-1, TGF-beta 3 may have the opposite effect. We hypothesize that the ratio of TGF-beta isoforms influences cutaneous scarring by altering the PAI-1:uPA activity ratio. Methods: The PAI-1/uPA activity in E14 TGF-beta 3 KO mice skins, skin fibroblasts and whole embryo lysates was studied using fibrin and reverse fibrin overlay. Additionally, an in vitro 3-D collagen gel model for wound contraction was employed to compare E14 TGF-beta 3 KO and wild-type (WT) fibroblasts. Results: E14 TGF-beta 3 KO skin, skin fibroblasts and embryo lysates had more PAI-1 and less uPA activity, compared to WT. Additionally, fibroblasts from E14 TGF-beta 3 KO mice contracted collagen gels less compared to WT, but could be rescued by adding TGF-beta 3. These fibroblasts also had a distinct morphology from WT. Conclusions: E14 TGF-beta 3 KO mice have a higher PAI-1:uPA ratio than WT. Furthermore, fibroblasts from this KO mouse are altered in matrix remodeling, which may provide a mechanism for the anti-scarring effects of TGF-beta 3.