Urokinase Plasminogen Activator Receptor and Soluble Matrix Metalloproteinase-9 in Acute Myeloid Leukemia Patients: A Possible Relation to Disease Invasion

Extracellular proteolytic enzymes of the urokinase-type plasminogen activator (uPA) and metalloproteinase (MMP) family play a crucial role in the matrix degradation and tissue remodeling process characteristic of malignant disorders. The receptor for urokinase plasminogen activator (uPAR) serves to localize and intensify the action of UPA and is expressed on the surface of malignant cells. Although the biological significance of MMP-9 and soluble urokinase receptor in growth and progression of lymphoid neoplasm is understood, its clinical significance in acute myeloid leukemia (AML) has not been fully elucidated. In this study, we determined the levels of soluble urokinase-type plasminogen activator receptor (suPAR), cellular uPAR and sMMP-9 in 43 newly diagnosed AML patients at diagnosis, before chemotherapy, and also studied 10 normal subjects served as a control group. After chemotherapy suPAR and MMP-9 were determined at remission and relapse. The levels of suPAR, cellular PAR were significantly higher (P=0.001, 0.001) and MMP-9 was significantly lower (P=0.001) in AML patients at diagnosis as compared to controls. suPAR and MMP-9 levels were significantly lower in AML patients who achieved complete remission (CR) as compared to those who did not (P=0.001 for both). Levels of suPAR and MMP-9 were significantly correlated to peripheral blood blast cells (r=0.88, P=0.001; r=0.65, P=0.001, respectively) and blast cell distribution ratio (BCDR, r=0.84, P=0.001; r=65, P=0.001, respectively). suPAR, cellular PAR and MMP-9 were significantly higher in patients with extramedullary infiltration as compared with those without (P=0.001, 0.001, <0.05). The suPAR, cellular uPAR, and MMP-9 levels were uneven in AML FAB subtypes being highest in M5(P<0.05 for all). MMP-9 and suPAR levels were correlated with the disease status. In AML survivors, MMP-9, cellular uPAR and suPAR were significantly lower as compared to non-survivors (P=0.001 for all).

In conclusion, MMP-9 and suPAR levels might be used as a marker for disease activity and may contribute to blast cell dissemination. MMP-9 and suPAR may be target molecules in the strategy of treatment of AML.     

AMI患者尿激酶与组织型纤溶酶原激活剂治疗结果的对比

比较31例尿激酶(UK)及14例组织型纤溶酶原激活剂(t-PA)静脉溶栓辅以阿斯匹林及肝素治疗急性心肌梗塞(AMI)的疗效.t-PA组与UK组相比较:血管再通率分别为78.6%与58.1%(P>0.05);脑、消化道及呼吸道出血并发症在t-PA组稍多,而UK组以局部皮肤出血较多.血管再通组心力衰竭、严重性心律失常、室壁瘤及梗塞后心绞痛的发生率较低,但两组间均无显著性差异;再通组病人心脏破裂的发生明显低于未再通组(0与17.6%P<0.05).本研究提示静脉t-PA溶栓治疗血管再通率高于静脉UK,有条件者可以首选t-PA.溶栓再通可以减少心力衰竭、室壁瘤、心梗后再缺血的发生,特别是心脏破裂的发生,从而改善病人的预后.     

Plasminogen activation in human acute leukaemias

Plasminogen activation is implicated in solid tumour growth, invasion and metatastic spread. However, little is known about its role in leukaemia. We investigated the production by leukaemic cells of plasminogen activators [urokinase (uPA) and tissue-type PA (tPA)], cell surface receptor for uPA (uPAR) and PA inhibitors (PAI-1 and PAI-2). Leukaemic cells from 37 patients [26 with acute myeloid leukaemia (AML) and 11 with acute lymphoid leukaemia (ALL)] were analysed for mRNA content and enzymatic activities. High levels of uPA mRNA were found in M1, M2, M3 and M4-M5 AMLs, whereas tPA mRNA was not detected in any of the analysed cases. uPAR mRNA was confined to subtypes M4-M5. PAI-1 mRNA was detected in M3 and M4-M5. PAI-2 mRNA was found predominantly in M2 and M4-M5. SDS-PAGE/zymography analyses of cell extracts and supernatants after 24 and 48 h of culture confirmed the production of active uPA by AML cells (mainly M4-M5). but not by ALL. The finding of uPA, uPAR, PAI-1 and PAI-2 synthesized by leukaemic cells suggests that plasminogen activation may contribute to the invasive behaviour of these cells, the fibrinolytic imbalance observed in leukaemic patients and the differentiation and proliferation of M4-M5 by interaction of uPA with uPAR.     

冠心病患者血浆脂蛋白(a)浓度与组织型纤溶酶原激活剂及抑制物活性的关系

为了了解冠心病患者血浆脂蛋白（a）浓度与组织型纤维蛋白溶解酶原激活剂及其抑制物活性的关系，进一步研究脂蛋白（a）促血栓形成的机理，本文应用酶联免疫吸附法测定20例冠心病患者与20名健康人脂蛋白（a）浓度，并以底物显色法测定其血浆组织型纤维蛋白溶解酶原激活剂及其抑制物活性。结果发现，冠心病组血脂蛋白（a）浓度和纤维蛋白溶酶原激活剂抑制物活性显著高于对照组，而纤维蛋白溶解酶原激活剂活性显著低于对照组；两组脂蛋白（a）浓度与纤维蛋白溶解酶原激活剂活性无关，而与纤维蛋白溶解酶原激活剂抑制物活性呈显著正相关。提示血浆高脂蛋白（a）浓度是冠心病的危险因素，冠心病患者纤溶与凝血功能失去平衡，促进了冠状动脉内血栓形成与动脉硬化的发展，进而导致和加重冠心病。     

冠心病患者血浆脂蛋白(a)浓度与组织型纤溶酶原激活剂及抑制物活性的关系

为了了解冠心病患者血浆脂蛋白(a)浓度与组织型纤维蛋白溶解酶原激活剂及其抑制物活性的关系,进一步研究脂蛋白(a)促血栓形成的机理,本文应用酶联免疫吸附法测定20例冠心病患者与20名健康人脂蛋白(a)浓度,并以底物显色法测定其血浆组织型纤维蛋白溶解酶原激活剂及其抑制物活性。结果发现,冠心病组血脂蛋白(a)浓度和纤维蛋白溶酶原激活剂抑制物活性显著高于对照组,而纤维蛋白溶解酶原激活剂活性显著低于对照组;两组脂蛋白(a)浓度与纤维蛋白溶解酶原激活剂活性无关,而与纤维蛋白溶解酶原激活剂抑制物活性呈显著正相关。提示血浆高脂蛋白(a)浓度是冠心病的危险因素,冠心病患者纤溶与凝血功能失去平衡,促进了冠状动脉内血栓形成与动脉硬化的发展,进而导致和加重冠心病。     

Urokinase type plasminogen activator receptor expression in colorectal neoplasms

Background—The urokinase type plasminogenactivator receptor (uPAR) may play a critical role in cancer invasionand metastasis.
Aims—To study the involvement of uPAR incolorectal carcinogenesis.
Methods—The cellular expression and localisationof uPAR were investigated in colorectal adenomas and invasivecarcinomas by in situ hybridisation, immunohistochemistry, and northernand western blot analyses.
Results—uPAR mRNA expression was found mainly inthe cytoplasm of dysplastic epithelial cells of 30% of adenomas withmild (19%), moderate (21%), and severe (47%) dysplasia, and in that of carcinomatous cells of 85% of invasive carcinomas: Dukes' stages A(72%), B (93%), and C (91%). Some stromal cells in the adjacent neoplastic epithelium were faintly positive. Immunoreactivity for uPARwas detected in dysplastic epithelial cells of 14% of adenomas and incarcinomatous cells of 49% of invasive carcinomas. uPAR mRNA andprotein concentrations were significantly higher in severe than in mildor moderate dysplasia (p<0.05); they were notably higher in Dukes'stage A than in severe dysplasia (p<0.05), and significantly higher inDukes' stage B than in stage A (p<0.05), but those in stage B werenot different from those in stage C or in metastatic colorectalcarcinomas of the liver.
Conclusions—Colorectal adenoma uPAR, expressedessentially in dysplastic epithelial cells, was upregulated withincreasing severity of atypia, and increased notably during thecritical transition from severe dysplasic adenoma to invasivecarcinoma. These findings implicate uPAR expression in the invasive andmetastatic processes of colorectal cancer.

Keywords:urokinase type plasminogen activator receptor; colorectal adenoma; colorectal cancer; adenoma-carcinoma sequence

     

单核细胞尿激酶型纤溶酶原激活物受体表达水平与冠状动脉粥样硬化斑块稳定性的关系

目的探讨不同临床类型的冠状动脉粥样硬化性心脏病患者外周血单核细胞尿激酶型纤溶酶原激活物受体(uPAR)表达水平的差异及其在治疗前后的变化,并观察该指标与术后心血管事件的相关性。方法连续入选经冠状动脉造影证实的冠心病患者136例,其中包括急性冠脉综合征(ACS)85例、稳定型心绞痛(SAP)51例,另选20例健康志愿者作为对照。留取全血标本,用流式细胞仪检测各组患者外周血单核细胞uPAR的表达水平。观察ACS组经皮冠状动脉介入(PCI)治疗24,48h,3个月后外周血单核细胞uPAR表达水平的变化。随访ACS组患者2年,记录主要心血管事件的发生情况。结果 ACS组uPAR表达水平较SAP组及健康志愿者组均显著升高(P0.01),ACS组PCI术后3个月的uPAR表达水平较术前有显著下降(P0.01)。85例ACS患者平均随访(20.46±2.45)个月,术后发生主要心血管事件24例,事件组单核细胞uPAR表达水平显著高于无事件组(P0.01)。应用多因素logistic回归分析发现,单核细胞上uPAR的表达水平与术后主要心血管事件显著相关(P0.01)。结论 ACS患者单核细胞uPAR的表达水平较SAP患者显著升高,ACS患者uPAR的表达水平与主要心血管事件具有明显相关性,单核细胞uPAR的表达水平是冠心病临床病情严重程度的一个敏感指标,对于判断动脉粥样斑块的稳定性以及心血管疾病的预后有重要价值。     

血清suPAR水平与慢性心力衰竭患者预后的关系研究

目的探讨血清可溶性尿激酶型纤溶酶原激活物受体(suPAR)与慢性心力衰竭(CHF)患者预后的关系。方法选取2016年1月至2017年12月期间于解放军联勤保障部队第九0九医院心血管内科收治的120例CHF患者作为CHF组(n=120),另选取同期来院进行体检的120例健康体检者作为对照组(n=120)。分析对照组及CHF组血清suPAR的表达情况,探讨其与炎症因子如肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和心功能指标如左室舒张末期内径(LVEDD)、左室射血分数(LVEF)的相关性,并随访两组研究对象12个月,分析血清suPAR对CHF患者预后的评估价值。结果CHF组的血清suPAR、TNF-α、IL-6、N末端脑钠肽前体(NT-proBNP)水平以及LVEDD均明显高于对照组(P<0.05),LVEF明显低于对照组(P<0.05)。CHF患者中,NYHA分级Ⅳ级、Ⅲ级、Ⅱ级患者的血清suPAR、TNF-α、IL-6、NT-proBNP水平以及LVEDD逐渐降低,LVEF逐渐升高,两两比较差异均有统计学意义(P<0.05)。CHF患者中,血清suPAR水平与TNF-ɑ、IL-6、LVEDD、NT-proBNP均呈正相关,与LVEF呈负相关(P<0.05)。随访12个月,共28例患者发生心脏事件。预后不良组患者的血清suPAR、TNF-α、IL-6、NT-proBNP水平以及LVEDD均明显高于预后良好组,LVEF明显低于预后良好组(P<0.05)。多因素Logistic回归分析显示:suPAR水平过高及LVEF过低均是CHF患者发生心脏事件的独立危险因素(P<0.05)。受试者操作特征曲线(ROC)分析显示:血清suPAR对CHF患者预后有较高的评估价值,曲线下面积为0.798,敏感度和特异性为75.00%和77.21%。结论血清suPAR在CHF患者中表达明显上升,且是CHF患者发生心脏事件的独立危险因素,对患者的预后评估具有较高的应用价值。     

Fibrinolytic characteristics and their significance in malignant, tuberculous and cirrhotic pleural and ascitic fluids

The fibrinolytic characteristics and clinical pathological significance of pleural and ascitic fluid were studied in patients with malignant tumour, tuberculosis or liver cirrhosis. Urokinase plasminogen activator (uPA) and urokinase plaminogen activator receptor (uPAR) levels were measured by enzyme-linked immunoadsorbent assay and tissue plasminogen activator (tPA), plasminogen activator inhibitor-1 (PAI-1), plasminogen (Plg), plasmin (Pl) and alpha(2) plasmin inhibitor (alpha(2)PI) by colorimetric assay. uPA and uPAR levels were higher in malignant tumour and tuberculosis compared with liver cirrhosis, whereas tPA levels were significantly higher in liver cirrhosis and malignant tumour than in tuberculosis patients. Tuberculosis patients showed statistically higher PAI-1, Plg and Pl concentrations than malignant tumour patients, which, in turn, were higher than those in liver cirrhosis patients. alpha(2)PI levels were markedly higher in malignant tumour and liver cirrhosis than in tuberculosis. In the malignant tumour group, only uPA level was significantly different between the samples that contained cancer cells and those that did not. We found significant differences between the fibrinolytic characteristics in pleural and ascitic fluid in patients with malignant tumour, tuberculosis and liver cirrhosis. The analysis of several fibrinolytic parameters could help to determine the quality of pleural and ascitic fluid, and also to further understand the pathological processes of these diseases.     

慢性乙型肝炎患者血清suPAR和IP-1O水平及其临床意义

目的 探讨以血清可溶性尿激酶型纤溶酶原激活物受体（suPAR）和干扰素γ诱导蛋白-10（IP-10）水平诊断慢性乙型肝炎肝纤维化的可能性。方法 65例接受肝组织病理学检查的慢性乙型肝炎患者,轻度肝纤维化组37例和重度肝纤维化组28例。另选择同期20例肝移植供肝者作为对照。采用ELISA检测血清suPAR和IP-1O水平。采用受试者工作特征曲线（ROC）分析suPAR和IP-10对慢性乙型肝炎患者重度肝纤维化的诊断价值。结果 慢性乙型肝炎患者血清suPAR水平为4.5（1.8~10.5） ng/ml）,显著高于健康人（2.2（0.7~4.4） ng/ml,P<0.05）;血清IP-10水平为255（71~455） pg/ml,显著高于健康人（87（70~135） pg/ml,P<0.05）;轻度纤维化患者血清suPAR和IP-10水平分别为3.5 （1.7~5.5） ng/ml和215 （68~408） pg/ml,显著低于重度纤维化患者的（5.2 （1.8~9.2） ng/ml和290 （106~525） pg/ml （P<0.05）;以血清suPAR>4.2 ng/ml 或IP-10>265 pg/ml为截断点,其诊断慢性乙型肝炎患者重度肝纤维化的曲线下面积（AUC）分别是0.742和0.698,其敏感性分别为67.86%和64.29%,特异性分别为78.34%和72.97%。结论 血清suPAR和IP-10可作为慢性乙型肝炎肝纤维化的生物标志物,有助于慢性乙型肝炎肝纤维化的早期诊断。     

氧化型低密度脂蛋白对单核细胞表达尿激酶型纤溶酶原激活物受体的影响

目的探讨不同浓度的氧化型低密度脂蛋白（oxLDL）对体外培养单核细胞表达尿激酶型纤溶酶原激活物受体（uPAR）的影响。方法采用密度梯度离心法及黏附法分离、提取并纯化健康人外周血单核细胞,对原代培养的单核细胞分别加入25,50,100mg/L浓度的oxLDL,分别培养12,24,48h,测定单核细胞的uPAR蛋白表达量及uPAR mRNA的水平变化。药物干预组加入含阿托伐他汀（终浓度为5.0μmo/L）和50mg/L的ox-LDL的培养液培养12,24,48h,同样方法测定单核细胞的uPAR蛋白表达量及uPAR mRNA的水平变化。结果与正常组比较,oxLDL刺激组单核细胞uPAR蛋白表达水平有显著升高,并呈剂量依赖关系;oxLDL刺激组单核细胞uPAR mRNA的合成量被显著上调。阿托伐他汀干预组uPAR蛋白的表达水平及uPAR mRNA合成量的刺激效果均被显著抑制。结论 oxLDL刺激单核细胞高表达uPAR是通过转录水平的上调来刺激蛋白质合成增加的;阿托伐他汀对uPAR表达的抑制是通过下调uPAR转录水平来实现的。     

Moxonidine Effect on Microalbuminuria, Thrombomodulin, and Plasminogen Activator Inhibitor-1 Levels in Patients with Essential Hypertension

Increased sympathetic activity seems to play an important role in the pathogenesis and development of complications of atherosclerotic origin in patients with essential hypertension (EH). The aim of this study was to evaluate the effect of a new antihypertensive agent, moxonidine (M), on microalbuminuria (urine albumin excretion, UAE), plasma thrombomodulin (TM), and tissue plasminogen activator inhibitor (PAI-1) in patients with mild to moderate EH associated with increased UAE. Fifty-eight patients (32 M, 26 F) with EH and microalbuminuria, with a mean age of 56.6 ± 8.2 years and a body mass index (BMI) of 23.8 ± 3.1 kg/m2 who responded to M therapy (0.3–0.4 mg/daily) were studied before and after their blood pressure control. The 24-hour urine albumin excretion (RIA method), as well as TM and PAI-1 plasma levels (ELISA method), were determined before and 6 months after the initiation of treatment under moxonidine therapy. At the end of the 6-month period, all patients remained normotensive. The 24-hour urine albumin excretion had decreased to 24.5 ± 6.4 vs. 32.3 ± 7.2 ug/min before therapy (P < 0.001). The plasma TM levels had decreased to 44.0 ± 7 vs. 51.0 ± 9 ng/mL before therapy (P < 0.01), and PAI-1 levels had also decreased to 11.5 ± 4.5 vs. 15.8 ± 8 IU/mL before therapy (P < 0.05). The results of our study suggest that in hypertensive patients with microalbuminuria, moxonidine, an imidazoline I1-receptor agonist, a new centrally acting antihypertensive agent, significantly reduces urine albumin excretion as well as thrombomodulin and PAI-1 levels. These preliminary findings demonstrate a favorable effect on renal function and endothelial homeostatic mechanisms (maintenance of haemostatic balance).     

Chromosomal localization of the human urokinase plasminogen activator receptor and plasminogen activator inhibitor type-2 genes: Implications in colorectal cancer

Abstract Activation of the proenzyme of urokinase (uPA) on the surface of cancer cells has been implicated in the initiation of focal proteolytic mechanisms that permit invasion and metastasis by colon cancers. The activity of uPA on the cell surface appears to be a function of the number of uPA-specific receptors (uPAR) and the extent of inhibition of uPA by plasminogen activator inhibitors (PAI). The mapping of the genes coding for uPAR, and for PAI-2, was performed to determine whether their chromosomal localization suggested their involvement in the genetic alterations associated with cancer cell DNA.
This study confirms the localization of the human urokinase plasminogen activator receptor gene to chromosome 19q and, using in situ hybridization, provides a precise localization to chromosome 19q13.2. In addition, our results confirm the previous allocation of the human plasminogen activator inhibitor-2 gene to a location 18q21.3 → 18q21.1, a location that corresponds to the commonest (>70%) somatic deletions found in colorectal carcinomas. The mapping of the uPAR and PAI-2 genes enables the elucidation of their possible involvement in the genetic alterations that determine the invasive and metastatic phenotypes in colorectal cancer.     

血清可溶性转铁蛋白受体水平与慢性阻塞性肺疾病患者认知障碍程度的相关性研究

背景目前,血清可溶性转铁蛋白受体(sTFR)水平与慢性阻塞性肺疾病(COPD)患者认知障碍的关系尚未完全明确。目的探讨血清sTFR水平与COPD患者认知障碍程度的相关性。方法选取2017年2月-2019年7月在西安交通大学第一附属医院就诊的COPD患者248例作为观察组,另选取同期健康体检者250例作为对照组;另根据贫血程度将COPD患者分为轻度组38例、中度组45例、重度组19例及非贫血组146例。比较对照组、观察组及轻度组、中度组、重度组、非贫血组患者血清sTFR、内皮素1 (ET-1)、铁调节素(hepcidin)、8-羟基脱氧尿苷酸(8-OHdG)、低氧诱导因子1α(HIF-1α)水平及蒙特利尔认知评估量表(MoCA)评分;血清sTFR水平、MoCA评分与COPD患者血清ET-1、hepcidin、8-OHdG、HIF-1α水平间的相关性分析及血清sTFR水平与COPD患者MoCA评分的相关性分析均采用Pearson相关分析。结果 (1)观察组患者血清sTFR、ET-1、hepcidin、8-OHdG、HIF-1α水平高于对照组,MoCA评分低于对照组(P<0.05)。(2)轻度、中度、重度组患者血清sTFR、ET-1、hepcidin、8-OHdG、HIF-1α水平高于非贫血组,MoCA评分低于非贫血组(P<0.05);中度、重度组患者血清sTFR、ET-1、hepcidin、8-OHdG、HIF-1α水平高于轻度组,MoCA评分低于轻度组(P<0.05);重度组患者血清sTFR、ET-1、hepcidin、8-OHdG、HIF-1α水平高于中度组,MoCA评分低于中度组(P<0.05)。(3) Pearson相关分析结果显示,血清sTFR水平与COPD患者血清ET-1 (r=0.482)、hepcidin(r=0.682)、8-OHdG(r=0.513)、HIF-1α(r=0.441)水平均呈正相关(P<0.05),MoCA评分与COPD患者血清ET-1 (r=-0.461)、hepcidin (r=-0.515)、8-OHdG(r=-0.485)、HIF-1α(r=-0.437)水平均呈负相关(P<0.05);血清sTFR水平与COPD患者MoCA评分呈负相关(r=-0.481,P<0.05)。结论随着贫血程度加重,COPD患者血清sTFR水平升高、认知障碍程度加重,且血清sTFR水平与COPD患者认知障碍程度有关。