尿激酶型纤溶酶原激活因子作用系统与肿瘤转移

尿激酶型纤溶酶原激活因子作用系统与肿瘤转移魏泓作者单位：第三军医大学实验动物学教研室（重庆６３００３８）纤溶酶原激活因子（ＰＡ）可分为尿激酶型（ｕＰＡ）及组织型（ｔＰＡ）两种。两者均具有水解纤溶酶原转化为纤溶酶的能力，其不同的是，ｔＰＡ与纤维蛋白有高...     

转化生长因子β1和丁酸钠对结肠癌细胞UPA和PAI-1调节作用的影响

研究表明 [1],尿激酶型纤溶酶原激活剂（ urokinase plasminogen activator, UPA）和纤溶酶原激活物抑制剂（ plasminogen activator inhibitor 1, PAI 1）与肿瘤浸润和转移密切相关。 UPA、 UPA受体（ UPA receptor, UPAR）、底物分子（纤溶酶原）、抑制因子（ PAI 1）组成的尿激酶分子系统,通过降解细胞外基质介导细胞转移在肿瘤发展中起重要作用。本研究将利用结肠癌 LoVo细胞株,探讨转化生长因子β 1(TGF β 1)和丁酸钠对肿瘤细胞尿激酶分子系统的调控作用及对细胞增殖的影响。     

尿激酶型纤溶酶原激活物系统在正常子宫内膜和子宫内膜癌中的表达

 尿激酶型纤溶酶原激活物系统包括尿激酶型纤溶原激活物 (Urokinase- type PlasminogenActivators,u PA)尿激酶型纤溶酶原激活物受体(Receptor for u PA,u PAR)、纤溶酶原激活物抑制剂 (Plasminogen Activator Inhibitor,PAI)。它们是体内生理或病理条件下通过降解胞外基质、介导细胞转移的主要酶系。其与肿瘤关系的研究近年来不断深入 ,本文将对尿激酶分子系统介导的肿瘤浸润与转移的机制 ,其在正常子宫内膜和子宫内膜癌中的表达等方面的进展作一综述。     

乳腺癌纤溶分子标志物t-PA、u-PA表达的临床研究

目的:检测乳腺癌患者血浆纤溶分子标志物含量、mRNA水平的变化以探索其与肿瘤浸润、播散的关系及临床意义。方法:采用逆转录、实时定量PCR技术检测30例乳腺癌组织中组织型纤溶酶原激活剂(tPA)、尿激酶型纤溶酶原激活剂(uPA)mRNA水平;用ELISA法同步检测患者血浆纤溶分子标志物含量,包括tPA、uPA、尿激酶型纤溶酶原激活剂受体(uPAR)、纤溶酶抗纤溶酶复合物(PAP)等。结果:乳腺癌患者血浆uPA、uPAR、PAP含量均较正常对照明显升高,其中,有周围淋巴结转移、远处脏器转移者uPA升高更为显著。肿瘤细胞uPAmRNA表达增高,在不同临床分期组中有显著差异,而tPAmRNA则减少。雌孕激素受体双阳性组tPAmRNA表达较其他组为高。结论:乳腺癌患者纤溶功能明显亢进,可能是其易播散、浸润的主要原因之一。逆转录实时定量PCR技术,使tPAmRNA、uPAmRNA有望成为临床上乳腺癌病情监测、预后判断及治疗选择的辅助指标。     

尿激酶纤溶酶原激活物系统与肿瘤侵袭转移

恶性肿瘤合成分泌大量胞外蛋白水解酶,降解细胞外基质及基底膜,是恶性肿瘤侵袭转移的重要基础.尿激酶纤溶酶原激活物(uPA)系统作为主要的蛋白水解酶在其中发挥重要作用.本简述uPA系统的结构、功能及其在侵袭、转移中的作用.     

纤溶酶原激活系统与肿瘤热化疗的关系

纤溶酶原的激活在肿瘤的侵袭、转移中发挥着重要作用，其基本理论是从肿瘤细胞分泌的纤溶酶原激活物催化水解无活性的纤溶酶原转变为有活性的纤溶酶，纤溶酶催化降解基底膜和细胞外基质(extracellular matrix，ECM)，有利于肿瘤细胞向周围组织侵袭。纤溶酶原激活物为丝氨酸蛋白酶，包括尿激酶型纤溶酶原激活物     

uPA,uPAR,PAI-1在具有不同转移率的鼻咽癌细胞系中的表达

目的：研究尿激酶型纤溶酶原激活剂（urokinase-type plasminogen activator,uPA)和它的受体（uPAR)、抑制剂（PAI－1）在具有不同转移率的鼻咽癌细胞系CNE－2Z（20％淋巴道转移）、CNE－2Z－H5（40％淋巴道转移）、CNE－2Z－H5－9（90％淋巴道转移）中的表达。方法：应用逆转录聚合酶反应（RT－PCR）方法检测uPA,uPAR,PAI-1在基因转录水平的表达。结果：uPA,uPAR mRNA在CNE－2Z－H5－9中表达最高，在CNE－2Z中表达最低；PAI－1 mRNA在CNE－2Z－H5－9中无表达，在CNE－2Z，CNE－2Z－H5有表达，但差异无显著性。结论：uPA、uPAR可能促进鼻咽癌细胞侵袭、转移，PAI－1可能起抑制作用。     

尿激酶型纤溶酶原激活剂在甲状腺癌发生发展及淋巴结转移中的作用

目的探讨尿激酶型纤溶酶原激活剂与AnnexinⅡ在甲状腺癌发生及淋巴结转移中的作用。方法使用免疫组化检测40例甲状腺乳头状癌、30例甲状腺腺瘤、10例结节性甲状腺肿、10例正常甲状腺组织中UPA蛋白及AnnexinⅡ的表达。结果甲状腺乳头状癌组织中UPA、AnnexinⅡ的表达显著高于结节性甲状腺肿及腺瘤组织,且与甲状腺癌的发生呈明显正相关;AnnexinⅡ表达与肿瘤大小、淋巴结转移有关,而UPA表达与淋巴结转移有关。结论 UPA、AnnexinⅡ蛋白表达与甲状腺乳头状癌的发生密切相关,且在甲状腺乳头状癌淋巴结转移过程中发挥重要作用。     

尿激酶型纤溶酶原激活因子及抑制剂表达与人大肠癌细胞转移能力的关系

目的　探讨尿激酶型纤溶酶原激活因子 ( u PA)、尿激酶型纤溶酶原激活因子受体 ( u PAR)和纤溶酶原抑制剂 1( PAI- 1)的表达与人大肠癌细胞系转移能力的关系。方法　用 EL ISA方法测定 3个人大肠癌细胞系培养上清液中 u PA、u PAR和 PAI- 1含量 ;用免疫组化 ABC方法检测 u PA、u PAR和 PAI- 1在细胞中的表达 ;分析其表达与大肠癌细胞转移能力的关系。结果　在培养上清液中具有高转移能力的 HT- 2 9d细胞的 u PA、u PAR及 PAI- 1含量明显高于低转移的 HT- 2 9和不转移的 Wi Dr细胞 ,而 HT- 2 9细胞的 u PA、u PAR及 PAI- 1含量则高于 Wi Dr细胞。免疫组化显示 u PA和 PAI- 1在 HT- 2 9d细胞中的表达高于 HT- 2 9和 Wi Dr细胞。结论　 u PA、u PAR和 PAI-1的表达与大肠癌细胞的转移能力密切相关。     

尿激酶型纤溶酶原激活物系列与肿瘤转移

尿激酶型纤溶酶原激活物(uPA)属丝氨酸蛋白水解酶，uPA与其受体(uPAR)、抑制剂(uPAI)所形成的作用系统可激活纤溶酶原参与细胞外基质的降解，介导肿瘤的侵袭和转移。现对uPA作用系统中的主要组分及其在肿瘤侵袭转移中作用作一简要介绍。     

Activities,localizations, and roles of serine proteases and their inhibitors in human brain tumor progression

Summary The plasminogen activation system consists of plasminogen activators and their inhibitors, serine proteases, and serpins. The proteases and inhibitors regulate a variety of processes in tissue morphogenesis, differentiation, cell migration, and cancer cell invasiveness and metastasis. One of the plasminogen activators, urokinase-type plasminogen activator (uPA), binds to a specific surface and provides a localized cell surface proteolytic activity required for the destruction of extracellular matrix, which is a vital step in tumor cell invasion. The proteolytic activity of uPA is modulated by its cell surface receptor, as well as by plasminogen activator inhibitor type-1 (PAI-1) and, to a lesser degree, by other inhibitors.The role of plasminogen activators and their inhibitors in cancer invasion can be demonstrated in the development and progression of malignant brain tumors. Our findings indicate that uPA and PAI-1 expression are dramatically upregulated in malignant brain tumors in parallel with the histological progression of the tumors. The results suggest that these molecules may contribute to tumor invasion in addition to their significant role in angiogenesis. An evaluation of the plasminogen activation system could add diagnostic and prognostic significance to the evaluation of individual patients.     

肝癌患者血浆、组织中凝血及纤溶因子的表达及其临床意义

背景与目的:研究表明凝血及纤溶因子改变与恶性肿瘤的发生、发展密切相关。本研究检测组织因子(tissuefactor,TF)、尿激酶型纤溶酶原激活物(urokinase-typeplasminogenactivator,uPA)及其受体(urokinase-typeplasminogenactivatorreceptor,uPAR)在肝细胞癌(hepatocellularcarcinoma,HCC)组织中及血浆中的表达并探讨其临床意义。方法:应用酶联免疫吸附法(ELISA)检测肝细胞癌患者50例及对照组30例非癌患者的血浆TF、uPA和uPAR水平;50例肝癌中随机选取27例取其肝癌组织、癌旁组织,以27例非癌患者肝组织为对照,利用RT-PCR法分别检测TF、uPA、uPARmRNA的阳性率及相对表达强度,并结合临床病理资料进行分析。结果:肝细胞癌患者血浆TF、uPA及uPAR水平均较对照组升高[(409.4±13.0)pg/mlvs.(318.8±69.1)pg/ml,(1.63±0.52)ng/mlvs.(1.20±0.40)ng/ml,(1.36±1.00)ng/mlvs.(0.68±0.28)ng/ml],均有显著性差异(P<0.05)。肝细胞癌患者血浆TF水平在低分化组、肿瘤较大组及合并肝硬化组均显著升高(P<0.05),血浆uPA水平只在合并肝硬化组升高(P<0.05)。肝细胞癌患者血浆TF、uPA和uPAR水平在有淋巴结转移、肝外脏器转移及门脉癌栓组较无转移及无癌栓组升高(P<0.05)。TF、uPA、uPARmRNA在肝细胞癌组织中阳性率及相对表达强度分别为62.96%(17/27)、70.37%(19/27)、77.78%(21/27)及0.57±0.27、0.96±0.46、0.78±0.32,均显著高于癌旁组织及非癌患者肝组织,均有显著性差异(P<0.05);TF、uPA、uPARmRNA在有肝内转移及门脉癌栓组的阳性率及相对表达强度均高于无肝内转移及门脉癌栓组(P<0.05)。经Pearson检验,肝细胞癌患者TF、uPA和uPARmRNA表达呈正相关(TF/uPA:r=0.37,P<0.01,TF/uPAR:r=0.53,P<0.01,uPA/uPAR:r=0.36,P<0.01)。经Cox多因素分析三者均为独立预后因素[TF(χ2=6.05,P=0.014),uPA(χ2=4.29,P=0.038),uPAR(χ2=4.40,P=0.036)]。结论:TF、uPA及uPAR可能在肝细胞癌的侵袭转移过程中起协同作用;三者可能与肝癌患者预后有关。     

CLINICAL STUDY OF T-PA AND U-PA EXPRESSION IN PATIENTS WITH GASTROINTESTINAL CANCER

Objective: To study the changes of tissue-type plasminogen activator (t-PA), urokinase-type plasminogen activator (u-PA) expressions and fibrinolysis molecular markers in patients with gastrointestinal cancer in order to elucidate their clinical significance. Methods: The plasma levels of t-PA, u-PA, urokinase-type plasminogen activator receptor (u-PAR) and plasmin anti-plasmin complex (PAP) were measured by ELISA. t-PA and u-PA mRNAs were detected by Real-time RT-PCR. Results: The plasma levels of u-PA, u-PAR and PAP were elevated in gastrointestinal cancer patients, while u-PA was markedly elevated in patients with local infiltration, lymph node involvement or distal metastasis. U-PA mRNA was higher and t-PA was lower in gastrointestinal cancer compared to normal tissue. Conclusion: Hyperfibrinolysis is an important factor related with metastasis potential of gastrointestinal cancer, t-PA may be a character of well differentiated tissue.     

Expression and significance of urokinase-type plasminogen activator in breast cancer

Themortalityofbreastcanceriscloselyrelatedtotheinvasiveandmetastaticpotentialofthecancercells.Fibrinolysismediatedbyurokinase-typeplasminogenactivator(uPA)candegradeextracellularmatrixandbasementmembrane.IncreasedlevelsofuPAcorrelatewithhigherinvasiveandmetastaticability.[']Inthispaper,westudiedtheexpressionofuPAinbreastcancertissueusingimmunohistochemicalmethod,combiningsuchexpressionwithfollow-updata,andexploringtherelationofsuchexpressiontoprognosis.MATERIALSANDMETHODSPatientsFro…     

Expression pattern of the urokinase-plasminogen activator system in rat DS-sarcoma: role of oxygenation status and tumour size

The urokinase plasminogen activator system plays a central role in malignant tumour progression. Both tumour hypoxia and enhancement of urokinase plasminogen activator, urokinase plasminogen activator-receptor and plasminogen activator inhibitor type 1 have been identified as adverse prognostic factors. Upregulation of urokinase plasminogen activator or plasminogen activator inhibitor type 1 could present means by which hypoxia influences malignant progression. Therefore, the impact of hypoxia on the expression pattern of the urokinase plasminogen activator system in rat DS-sarcoma in vivo and in vitro was examined. In the in vivo setting, tumour cells were implanted subcutaneously into rats, which were housed under either hypoxia, atmospheric air or hyperoxia. For in vitro studies, DS-sarcoma cells were incubated for 24 h under hypoxia. Urokinase plasminogen activator and urokinase plasminogen activator-receptor expression were analysed by flow cytometry. Urokinase plasminogen activator activity was measured using zymography. Plasminogen activator inhibitor type 1 protein levels in vitro and in vivo were examined with ELISA. PAI-1 mRNA levels were determined by RT-PCR. DS-sarcoma cells express urokinase plasminogen activator, urokinase plasminogen activator-receptor, and plasminogen activator inhibitor type 1 in vitro and in vivo. The urokinase plasminogen activator activity is enhanced in DS-sarcomas compared to normal tissues and rises with increasing tumour volume. The oxygenation level has no impact on the urokinase plasminogen activator activity in cultured DS-sarcoma cells or in solid tumours, although in vitro an increase in plasminogen activator inhibitor type 1 protein and mRNA expression after hypoxic challenge is detectable. The latter plasminogen activator inhibitor type 1 changes were not detectable in vivo. Hypoxia has been demonstrated to contribute to the upregulation of some components of the system in vitro, although this effect was not reproducible in vivo. This may indicate that the serum level of plasminogen activator inhibitor type 1 is not a reliable surrogate marker of tumour hypoxia.     

UTI对人膀胱癌T24细胞尿激酶型纤溶酶原激活剂表达的影响

 目的　体外研究尿胰蛋白酶抑制物对人膀胱癌T2 4细胞尿激酶型纤溶酶原激活剂表达的影响 ,探讨UTI抑制膀胱肿瘤侵袭和转移的机制。方法　采用免疫组化和半定量逆转录聚合酶链式反应(RT PCR)检测不同量的UTI对体外培养的人移行细胞膀胱癌T2 4细胞uPA表达的影响。结果　免疫组化和RT PCR结果均表明 ,UTI能拮抗uPA的表达并呈负的剂量效应关系。结论　UTI能有效抑制人膀胱癌T2 4细胞uPA的表达 ,为UTI抑制膀胱肿瘤侵袭和转移提供了实验依据。     

阿米洛利对高转移肺癌细胞侵袭能力和uPA 系统的影响*

目的:观察阿米洛利对人高转移肺癌细胞 PGCL3 体外侵袭能力和尿激酶型纤溶酶原激活物（urokinase-type plasminogen activator,uPA ）系统的影响。方法:不同浓度（25 μmol/L、50 μmol/L 和100μmol/L）的阿米洛利作用于PGCL3 细胞6h 后,用Transwell 小室法检测对细胞侵袭能力和运动能力的影响。阿米洛利作用 24 h 后,用发色底物法检测对细胞分泌的 uPA 和纤溶酶原激活物抑制剂-1（plasminogen activator inhibitor- 1,PAI-1）活性的变化。RT-PCR 检测阿米洛利对细胞uPA 、尿激酶型纤溶酶原激活物受体（urokinase-type plasminogen activator receptor ,uPAR）和PAI-1 mRNA 表达的影响。Western blot 检测阿米洛利对细胞uPA 、细胞外调节蛋白激酶2（extracellular regulated protein kinase 2,ERK 2）和ras蛋白表达情况的影响。结果:侵袭实验和运动实验结果均显示,经
阿米洛利处理后,穿膜细胞数明显减少,在100μmol/L 时,对侵袭和运动的抑制率分别为（37 .7±4.1）%和（64 .9±4.9）%,与对照组相比具有显著性差异（P<0.01 ）。同时,细胞分泌的 uPA 和PAI-1 活性降低,当浓度达到 100μmol/L 时,差异具有统计学意义（P<0.05 ）。从25 μmol/L 开始,阿米洛利就可显著抑制PAI-1 mRNA 的表达,当达 100μmol/L 时可明显下调uPA mRNA 的表达,但各浓度对uPAR mRNA的表达均无影响。随着阿米洛利浓度的增加,uPA 蛋白表达量逐渐减少,但对 ras和ERK 2 蛋白表达量无明显影响。结论:阿米洛利能够抑制高转移肺癌细胞PGCL3 的侵袭和运动,其作用机制与抑制uPA 和PAI-1 的活性和表达有关,有成为抗肿瘤转移药物的潜力。      

Urokinase plasminogen activator (uPA) and plasminogen activator inhibitor type-1 (PAI-1) in breast cancer - correlation with traditional prognostic factors

Background

Urokinase plasminogen activator (uPA) and plasminogen activator inhibitor type-1 (PAI-1) play a key role in tumour invasion and metastasis. High levels of both proteolytic enzymes are associated with poor prognosis in breast cancer patients. The purpose of this study was to evaluate the correlation between traditional prognostic factors and uPA and PAI-1 expression in primary tumour of breast cancer patients.

Patients and methods.

606 primary breast cancer patients were enrolled in the prospective study in the Department of gynaecological oncology and breast oncology at the University Medical Centre Maribor between the years 2004 and 2010. We evaluated the traditional prognostic factors (age, menopausal status, tumour size, pathohistological type, histologic grade, lymph node status, lymphovascular invasion and hormone receptor status), together with uPA and PAI-1. We used Spearman’s rank correlation, Mann Whitney U test and χ² test for statistical analysis.

Results

Our findings indicate a positive correlation between uPA and tumour size (p < 0.001), grade (p < 0.001), histological type (p < 0.001), lymphovascular invasion (p = 0.01) and a negative correlation between uPA and hormone receptor status (p < 0.001). They also indicate a positive correlation between PAI-1 and tumour size (p = 0.004), grade (p < 0.001), pathohistological type (p < 0.001) and negative correlation between PAI-1 and hormone receptor status (p = 0.002).

Conclusions

Our study showed a relationship between uPA and PAI-1 and traditional prognostic factors. Their role as prognostic and predictive factors remains to be further evaluated.     

Is there a role for urokinase-type plasminogen activator inhibitors as maintenance therapy in patients with ovarian cancer?

There is abundant evidence that the urokinase-type plasminogen activator (uPA), its inhibitors PAI-1 and PAI-2 (plasminogen activator inhibitor type-1 and type-2) and its cells surface receptor (uPA-R, CD87) play a fundamental role in tumor invasion and metastasis and are of significant prognostic significance for many tumor types. We performed a systematic Med-line search on uPA, PAI, uPA-R and (epithelial) ovarian cancer (EOC). The majority of malignant EOC specimens show moderate to strong immunostating of tumor and stromal cells. Overexpression of u-PA and PAI-1 can be found in more the 75% of primary ovarian carcinomas, in most metastatic EOC samples and all examined epithelial ovarian cancer cell lines. uPA overexpression in primary specimens was significantly associated with tumor stage, grade, residual disease status after cytoreductive surgery, and poor clinical outcome. This may be explained by increased chemoresistance, a lower resectability and more aggressive tumor biology and tumor dissemination in patients with high uPA and PAI-1. Several therapeutical approaches aimed at inhibiting the uPA/uPAR functions have shown to possess anti-tumor effects in vitro and in animal models. When treating a patient with advanced ovarian cancer it may to be assumed that inhibiting the progression of established (micro) metastases may be more therapeutically relevant than trying to destroy all tumor cells which is not possible in most cases with current systemic treatment modalities. Taking into account the role of uPA and PAI in cell detachment, formation of new stroma, tumor cell reimplantation and metastasis uPA inhibition should be further investigated as maintenance treatment in patients with advanced EOC.