纤溶酶原激活因子及抑制因子与卵巢癌浸润转移

肿瘤浸润转移必须突破其周围的细胞外基质(ECM).纤溶酶原激活因子(PA)及其抑制因子(PAIs)介导的ECM的降解在卵巢癌的浸润转移过程中起十分关键的作用,肿瘤细胞通过大量表达uPA、PAI-1使ECM分解,进而不断浸润和扩散.对其基因结构、功能、作用机制等进行综述,旨在为肿瘤的诊断和治疗提供新方法,开辟新途径.     

纤溶酶原激活因子及抑制因子与卵巢癌浸润转移

肿瘤浸润转移必须突破其周围的细胞外基质(ECM)。纤溶酶原激活因子(PA)及其抑制因子(PAIs)介导的ECM的降解在卵巢癌的浸润转移过程中起十分关键的作用，肿瘤细胞通过大量表达uPA、PAI-I使：ECM分解，进而不断浸润和扩散。对其基因结构、功能、作用机制等进行综述，旨在为肿瘤的诊断和治疗提供新方法，开辟新途径。     

纤溶酶原激活因子及抑制因子在胎膜组织中免疫定位的研究

胎膛由羊膜、绒毛膜和蜕膜组成，从形态上可分为八层[1]。它对胎儿在宫内生长和发育起重要保护作用。胎膜异常导致流产，引发胎儿和母体病变，甚至引起死亡。有证据表明，先兆性子痫或胎儿宫内生长迟缓伴随血液中纤溶酶原激活因子（PA）和抑制因子（PAI）含量的变化[2,3]。由此推测，由PA－PAI系统局部介导的细胞外基质降解可能对维持胎盘的正常功能和羊膜的破裂机制密切有关。本文用免疫萤光和激光共聚焦扫描显微镜研究了tPA、PAI－1和PAI－2在人的分娩胎膜组织中的定位。材料和方法一、胎膜组织制备分娩胎盘取自北京市海淀区中关…     

人精子中尿激酶型纤溶酶原激活因子及其受体的分布

本文采用免疫组化和免疫电镜法研究了正常健康男子精子中尿激酶型纤溶酶原激活因子（ｕＰＡ）及其受体（ｕＰＡＲ）的分布。结果表明ｕＰＡ主要分布在顶体内外膜和头部浆膜上；精子尾部浆膜也含有ｕＰＡ；在ｕＰＡ出现的部位同时也发现有ｕＰＡＲ存在。推测结合在精子受体膜上的ｕＰＡ可能在精子的运行、精液的液化和受精过程中发挥重要的作用。     

催乳素延缓hCG诱发大鼠排卵和下调纤溶酶原激活系统在卵巢中的表达

给幼龄大鼠注射10IU PMSG,48h后注射7IU hCG,或者hCG加不同剂量的催乳素(PRL).在不同时间取出卵巢,检查输卵管中卵子数和卵巢不同细胞中组织型纤溶酶原激活因子(tPA)和抑制因子(PAI-1)mRNA含量和活性.结果表明:PRL减少hCG诱发的排卵数并有明显剂量和时间抑制曲线.当hCG注射24h后,在两组动物输卵管的卵子数无明显差异.PRL同时抑制hCG所诱发的颗粒细胞tPA表达;与少匕相反PRL还能显著刺激膜一间质细胞PAI-1mRNA表达.上述实验结果表明,PRL只暂时延缓而不是完全抑制hCG诱发的大鼠排卵.上述作用可能是通过抑制纤溶酶激活系统在卵巢中的表达引起的.     

人受精卵和未受精卵培养液中尿激酶型纤溶酶原激活因子的含量比较

本研究采用ELISA双抗夹心法测定了15例患不育症妇女25个受精卵和18个未受精卵培养液中尿激酶型纤溶酶原激活因子(uPA)的含量.实验结果表明:受精卵培养液中uPA的含量为(530±546)IU/L,未受精卵培养液中uPA含量为(22O±55)IU/L,P<0.05,两者有显著性差异.提示uPA在精卵受精过程中可能有益于精子穿透卵子的透明带.提高受精成功率.     

纤溶酶原激活物抑制因子1基因启动子区4G和5G多态基因型分布及其与多囊卵巢综合征的相关性研究

目的探讨纤溶酶原激活物抑制因子1（PAI-1）基因启动子区4G及5G的多态基因型分布,及其与多囊卵巢综合征（PCOS）发病的关系。方法应用聚合酶链反应-限制性片段长度多态性技术,检测101例PCOS患者（PCOS组）与42例因男性或单纯输卵管因素不孕患者（对照组）PAI-1基因启动子区4G基因型及5G多态基因型分布,测定并计算两组患者的体重指数、胰岛素抵抗指数、胰岛素敏感指数及自然流产数等临床指标。根据体重指数将PCOS患者分为肥胖者（48例）与非肥胖者（53例）。结果PCOS组PAI-1基因启动子区4G基因型频率为57％（58／101）,对照组为38％（16／42）;PCOS组5G基因型频率为43％（43／101）,对照组为62％（26／42）。两组4G及5G基因型频率分别比较,差异均有统计学意义（P〈0．05）。肥胖者胰岛素抵抗指数、胰岛素敏感指数与非肥胖者比较,差异均有统计学意义（P〈0．05,P〈0．01）。肥胖者4G基因型频率为48％（23／48）,非肥胖者为68％（36／53）;肥胖者5G基因型频率为52％（25／48）,非肥胖者为32％（17／53）。两者4G及5C基因型频率分别比较,差异均有统计学意义（P〈0．05）。27例有妊娠史的PCOS患者中,自然流产14例,4G基因型频率为79％（11／14）,5G基因型频率为21％（3／14）;无自然流产13例,4G基因型频率为38％（5／13）,5G基因型频率为62％（8／13）,两者4G与5G基因型频率分别比较,差异均有统计学意义（P〈0．05）。结论PAI-1基因启动子区4G基因型频率高,可能与PCOS的发病,尤其是与非肥胖者发生PCOS有关,并与PCOS患者的自然流产率高有关。     

尿激酶型纤溶酶原激活系统与卵巢肿瘤及其治疗

尿激酶型纤溶酶原激活系统是纤溶系统的重要组成部分,其通过水解细胞外间质,参与组织改造和细胞迁移,在肿瘤细胞的侵袭和转移中发挥作用。许多研究发现,尿激酶型纤溶酶原激活系统与卵巢癌恶性程度、分期、分化、转移及预后有重要关系。抑制尿激酶型纤溶酶原激活剂(uPA)的活性、阻断尿激酶型纤溶酶原激活与其受体在细胞表面结合、抑制信号传导途径等可作为卵巢肿瘤治疗的另一途径。综述尿激酶型纤溶酶原激活系统组成、作用机制,与卵巢肿瘤及其治疗研究进展。     

尿激酶型纤溶酶原激活系统与卵巢肿瘤及其治疗

尿激酶型纤溶酶原激活系统是纤溶系统的重要组成部分,其通过水解细胞外间质,参与组织改造和细胞迁移,在肿瘤细胞的侵袭和转移中发挥作用.许多研究发现,尿激酶型纤溶酶原激活系统与卵巢癌恶性程度、分期、分化、转移及预后有重要关系.抑制尿激酶型纤溶酶原激活剂(uPA)的活性、阻断尿激酶型纤溶酶原激活与其受体在细胞表面结合、抑制信号传导途径等可作为卵巢肿瘤治疗的另一途径.综述尿激酶型纤溶酶原激活系统组成、作用机制,与卵巢肿瘤及其治疗研究进展.     

Alteration of human follicular fluid plasminogen activator activity by ovarian hyperstimulation

Recently, it has been shown that granulosa-cell secretion of plasminogen activator (PA) is responsive to luteinizing hormone (LH)/human chorionic gonadotropin (hCG) as well as follicle stimulating hormone (FSH) in the rat and the pig. Accordingly, we asked whether PA activity in follicular fluid from exogenously stimulated human follicles was different from that of normal cycles and whether or not these activities correlated with follicular maturation as determined by follicular fluid steroid concentration. Follicular aspirates were obtained from women who were participating in an in vitro fertilization protocol. Follicular fluid concentrations of estradiol and progesterone were determined by established radioimmunoassay. PA activity, determined using a modified indirect solid-phase radioassay, was significantly less in follicles from patients treated with human menopausal gonadotropin (hMG) plus clomiphene (P<0.05) compared to untreated patients or those receiving hMG or clomiphene alone. Correlations of PA activity and follicular fluid steroid concentrations demonstrated no significant correlation in samples from treated patients. In contrast, untreated spontaneously cycling patients had a significant (r=0.89,P<0.05), positive correlation between follicular fluid estradiol levels. There was no correlation between PA activity and follicular fluid progesterone levels in any of the groups. These results suggest that a subtle balance in granulosa-cell secretion of PA and steroids exists, which appears to be disrupted by follicular hyperstimulation during treatment of patients participating in in vitro fertilization protocols.Supported in part by Clinical Investigator Award HD-00401 and by the University of Southern California Faculty Research and Innovation Award.     

子宫内膜癌患者血清uPA和PAI-1的含量变化及临床意义

目的:探讨子宫内膜癌患者血清尿激酶型纤溶酶原激活物(uPA)及纤溶酶原激活物抑制物-1(PAI-1)含量的变化及其临床意义。方法:用ELISA法测定35例子宫内膜癌(内膜癌组)、18例子宫内膜增生(内膜增生组)和16例正常子宫内膜患者(正常对照组)血清uPA和PAI-1含量及计算两者的比值。结果:内膜癌组患者血清uPA和PAI-1含量及uPA/PAI-1值均显著高于内膜增生组及正常对照组,差异有极显著性(P均<0.01)。内膜增生组及正常对照组,差异无统计学意义(P>0.05)。内膜癌组Ⅲ~Ⅳ期患者血清uPA和PAI-1含量与Ⅰ期相比差异有极显著性(P<0.01),Ⅲ~Ⅳ期患者血清uPA和PAI-1含量高于Ⅱ期(P<0.05),Ⅱ期患者血清uPA、PAI-1含量高于Ⅰ期(P<0.05)。内膜癌组患者血清uPA、PAI-1含量及uPA/PAI-1值随着手术病理分期及组织学分级的增高、肌层浸润深度的增加及淋巴结的转移而升高,差异有显著性(P均<0.05),而与患者病理类型无关(P>0.05)。结论:子宫内膜癌患者血清uPA和PAI-1含量及uPA/PAI-1值明显升高,并与其手术病理分期、浸润转移有关,提示其可能在内膜癌的发生、发展及浸润过程中起重要作用。     

尿激酶型纤溶酶原激活因子介导的卵巢上皮癌细胞浸润转移体外的实验研究

目地:探讨尿激酶型纤溶酶原激活因子在卵巢上皮癌浸润转移中的作用机制。方法:用RT-PCR技术从人卵巢上皮癌组织总RNA中逆转录uPA基因cDNA全长,克隆至pGEM-T Easy Vector,鉴定后与真核表达载体PCMV-HA连接,酶切及测序。用脂质体法将重组质粒DNA转染至SKOV3细胞。加压筛选并培养PCMV-HA-uPA及对照细胞。分别用RT-PCR和W estern blot方法检测转染前后SKOV3细胞uPA的表达。细胞增殖能力测定用四甲基偶氮唑蓝(MTT)法和集落形成实验,细胞周期测定用流式细胞仪法,细胞体外侵袭,迁移和黏附能力测定分别采用Matrigel Invasion,TranswellM igration和Adhersion Assay方法。结果:(1)uPA阳性表达能介导SKOV3细胞克隆形成,与对照组细胞的差异有统计学意义(P<0.05);(2)uPA阳性表达能介导SKOV3细胞的细胞周期中S期比例增加,与对照组细胞的差异有统计学意义(P<0.05);(3)uPA阳性表达介导SK-OV3细胞的体外侵袭,迁移和黏附能力均明显强于对照细胞株,差异有统计学意义(P=0.0002,<0.0001和0.0049)。结论:uPA通过促进肿瘤细胞侵袭,迁移和黏附能力在卵巢上皮癌浸润转移中起了重要作用。     

Plasminogen activator/plasminogen activator inhibitor-1 and cytokine modulation by the PROACT System

OBJECTIVE: To examine the effects of the PROACT treatment on the fibrinolytic system and inflammatory cytokines in human peritoneum. DESIGN: Controlled clinical study. SETTING: University hospital. PATIENT(S): Nine subjects undergoing laparotomy had peritoneal samples taken at the incision. INTERVENTION(S): The PROACT applicator was inserted through the peritoneal incision, and treatment of peritoneum was performed twice. A peritoneal sample was taken from one treated area. At closure, the second treated sample and an additional control sample were taken. All four samples were snap frozen in liquid nitrogen. Samples were homogenized and protein content extracted. MAIN OUTCOME MEASURE(S): Concentrations of total and active transforming growth factor-beta 1 (TGF-beta1), tumor necrosis factor-alpha (TNF-alpha), tissue-type plasminogen activator (t-PA), urokinase plasminogen activator (uPA), and plasminogen activator inhibitor 1 (PAI-1) were obtained. RESULT(S): Total TGF-beta1 at opening was 30% less in treated samples. At closure, active TGF-beta1 increased significantly (163%) in control samples and not in treated samples. Tumor necrosis factor alpha was detectable only in control samples at closure. During surgery, tPA levels showed a marked decrease in control samples vs. a small increase in treated samples. Levels of uPA increased significantly only in the control samples. In control samples, tPA/PAI-1 ratio was two thirds of treated sample ratio. CONCLUSION(S): Heating of the peritoneum with the PROACT System modulates the biologic tissue response to induce effects that would be consistent with inhibition of postoperative adhesion development.     

Soluble urokinase plasminogen activator receptor in preoperatively obtained plasma from patients with gynecological cancer or benign gynecological diseases

OBJECTIVE: The present study was planned to measure preoperative levels of soluble urokinase plasminogen activator receptor (suPAR) in plasma from patients with gynecological diseases, and to test for a relationship to clinical and biochemical patient characteristics. METHODS: Using a specific and sensitive kinetic ELISA, suPAR levels were determined in preoperative citrate plasma samples from 53 ovarian, 34 endometrial, and 30 cervical cancer patients, 17 patients with benign ovarian tumors, and 28 patients with benign endometrial diseases. In addition, suPAR was measured in citrate samples from 31 female blood donors. RESULTS: suPAR was measurable in all samples. No significant difference was found between plasma suPAR in the blood donors and the patients with benign diseases (P = 0.58). The groups of cancer patients had suPAR levels that were significantly higher than those found in the blood donors (P < 0.0001, P < 0.0001, and P = 0.001 for patients with ovarian, endometrial, and cervical cancer, respectively). In all groups of cancer patients a trend toward increasing suPAR levels with increasing FIGO stage was noted (P = 0.0003, P = 0.02, and P = 0.01 for patients with ovarian, endometrial, and cervical cancer, respectively). Using the median suPAR level to dichotomize the ovarian cancer patients, FIGO stages I-III, a significantly increased risk of progression/relapse was found for patients with high suPAR levels (Hazard ratio (HR) = 3.1, 95% CI: 1.1-8.8, P = 0.03). A multivariate analysis was performed, including suPAR, FIGO stage, and CA-125. Only FIGO stage III compared with FIGO stage I was significant (HR = 15, 95% CI: 1.8-129, P = 0.01). Survival analyses were not performed in the endometrial or cervical cancer patients due to few progressions/relapses during the follow-up period. CONCLUSION: This study concludes that patients with gynecological cancers have elevated plasma suPAR levels as compared with healthy female blood donors and patients with benign gynecological diseases. In addition, high preoperative plasma levels of suPAR are significantly associated with poor outcome of ovarian cancer patients. However, additional studies are needed to further validate the clinical usefulness of plasma suPAR measurements in the management of ovarian cancer patients.