폐암 환자에서 Urokinase Type Plasminogen Activator 및 Plasminogen Activator Inhibitor Type 1의 임상적 의의

권순대,최원일,한승범,권건영,전영준 계명대학교 의과학연구소 2000 계명의대학술지 Vol.19 No.2

Cancer invasion and metastasis require the dissolution of extracellular matrix in which several proteolytic enzyme are involved. One of these enzyme the urokinase-type plasminogen activator(u-PA), and plasminogen activator inhibitor type 1(PAI-1) have a possible role in cancer invasion and metastasis by protection of cancer itself from proteolysis by u-PA. It has been reported that the level of u-PA and PAI-1 in various cancer tissues are significantly higher than those in normal tissue and significant correlation with tumor size and lymph node involvement. We measured the concentration of plasma u-PA and PAI-1 antigens in patients with lung cancer and compared the concentration of them with histologic types and staging parameters, and also compared those concentrations in pre-treatment and post-treatment. In this study we measured the concentration of plasma u-PA and PAI-1 antigens using commercial ELISA kit in 40 lung cancer patients and 22 patient with benign lung diseases. The concentration of u-PA was 1.37±0.7 ng/mL in a group of benign lung disease patients and 1.75±0.75 ng/mL in lung cancer patients. The concentration of PAI-1 was 20.86±13.2 ng/mL in benign lung disease and 20.09 ng/mL in lung cancer. The concentration of u-PA in lung cancer patients was higher than those of benign lung disease patients. The concentration of u-PA was 2.42±2.69 ng/mL in lung cancer patients who were not treated, 1.78 ng±0,79 ng/mL in patients who were treated. The concentration of PAI-1 was 19.53±11.75 ng/mL in not-treated lung cancer patients, 10.71±6.26 ng/mL in treated patient group. The concentration of PAI-1 in treated lung cancer patients was lower than those of not-treated lung cancer patients. The concentration of u-PA was 1.82 ng/mL in stage I & Ⅱ, 1.93±0.11 ng/mL in stage Ⅲ, 1.65±0.17 ng/mL in stage Ⅳ. The concentration of PAI-1 was 15.92±5.57 ng/mL in stage I & Ⅱ, 20.95±0.54 ng/mL in stage Ⅲ, 23.99±2.5 ng/mL in stage Ⅳ. The concentration of u-PA was 1.28±0.45 ng/mL in small cell carcinoma, 1.86±0.12 ng/mL in nonsmall cell carcinoma 1.76±0.0 ng/mL in squamous cell carcinoma 1.93±0.2 ng/mL in adenocarcinoma. The concentration of PAI-1 was 18.74±3.83 ng/mL in small cell carcinoma 23.13±3.95 ng/mL in nonsmall cell carcinoma 25.39±2.81 ng/mL in squamous cell carcinoma 20.96±1.62 ng/mL in adenocarcinoma. The plasma levels of u-PA in lung cancer patients were higher than those benign lung disease and plasma level of PAI- 1 in who were treated (surgery, chemotherapy and/or radiotherapy) were lower than those who were not treated. These findings suggest involvement of U-PA with local invasion of lung cancer and possible roles in predicting the prognosis and survival of lung cancer patients.

체외배양한 인제대정맥혈관내피세포에서 R. tsutsugamushi 감염이 Tissue Factor와 Type 1 Plasminogen Activator Inhibitor의 발현에 미치는 영향에 관한 연구

김미란,기선호,배현주,장우현,박선양,최강원 대한감염학회 1995 감염 Vol.27 No.4

목적:쯔쯔가무시질환시 생기는 전신혈액응고장애의 병인 기전이 rickettsia의 침투에 의한 혈관내피세포의 손상, 그에 따른 tissue factor의 발현, 뒤이은 tissue type plasminogen activator(tPA)의 분비와 보상기전으로 type 1 plasminogen activator inhibitor(PAI-1)이 분비되는 과정으로 생각하고 이를 보고자 연구를 시행하였다. 방법:체외배양한 혈관내피세포에 순수분리한 Rickettsia tsutsugamushi를 감염시킨후 상층액에서 ELISA법으로 tPA와 PAI-1을 측정하였고 혈관내피세포 단층배양에서 면역형광법으로 tissue factor를 측정하였으며 PAI-1 유전자의 발현을 확인하고자 Northern blot analysis로 PAI-1 mRNA를 확인하였다. 결과: 1) PAI-1 R. tsutsugamushi를 감염시킨후 24시간에 가장 높은 농도를 보이며 그 증가량은 정상대조군에 비해 2.5배에서 4.7배까지 증가 하였다. 2) R. tsutsugamushi를 감염시킨후 혈관내피세포에서 분비되는 tPA의 분비는 정상대조군에 비해 유의한 차이를 보이지 않았다. 3) Northern blot analysis에 의한 PAI-1 mRNA의 발현 검색 결과 정상대조군에 비해 R. tsutsugamush가 감염된 혈관내피세포에서는 PAI-1의 발현이 2.5배 정도 증가하였다. 4) 혈관내피세포 단층배양에 R. tsutsugamushi를 감염시킨후 24시간에 tissue factor단일클론 항체와 FITC-conjugated anti mouse IgG를 이용한 간접 면역형광항체법으로 tissue factor를 측정한 결과 혈관내피세포 표면에서 tissue factor의 존재를 확인할 수 있었다. 결론:단층배양한 혈관내피세포에 R. tsutsugamushi를 감염시켰을 때 tissue factor가 발현되었고 PAI-1의 분비가 증가하여 24시간에 가장 많이 분비되었다. 그러나 tPA의 분비는 정상대조군에 비해 유의한 차이를 보이지 않았다. Nothern blot analysis를 통한 PAI-1 mRNA의 발현 검색 결과 PAI-1이 새로이 생성되어 분비됨을 확인하였다. Background:Tissue type plasminogen activator(tPA), type 1 plasminogen activator inhibitor (PAI-1), and von Willebrand factor are known to be released into the sera of patients in disseminated intravascular coagulation(DIC). The main pathologic mechanism of tsutsugamushi disease is the vasculitis by direct endothelial cell invasion by R. tsutsugamushi which dosen't have endotoxin. It is suspected that the mechanisms of DIC and activation of plasminogen activation system are different from those of sepsis by other organisms. which is caused by endotoxin. We suspect that direct rickettsial invasion of endothelial cells causes endothelial cell damage, tissue factor release, which is followed by DIC, and tPA and PAI-1 are released as compensatory mechanism. Methods:We cultured endothelial cells from human umbilical cord vein, infected them with purified R. tsutsugamushi Gilliam strain, checked tPA and PAI-1 by ELISA in culture supernatant. Then we observed the tissue factor expression on cultured endothelial cell monolayer by indirect IF stain. PAI-1 gene expression was evaluated by northern blot analysis. Results: 1) PAI-1 level showed gradual increase up to 240ng/ml (2.5-4.7 fold increase) in 24 hours. 2) tPA level showed no significant change with time. 3) PAI-1 gene expression increased 2.5 fold by northern blot analysis. 4) Tissue factor was expressed on the endothelial cells infected with R. tsutsugamushi. Conclusion: R. tsutsugamushi infection induces expression of tissue factor on endothelial cells and PAI-1 synthesis and it would contribute to DIC mechanism in tsutsugamushi disease in part. But it has no direct effect on tPA release.

위암의 침윤과 전이에 관여하는 단백분해 효소 ( uPA , PAI - 1 및 Type 4 Collagenase ) 에 관한 연구

조재용(J . Y . Cho),정현철(H . C . Chung),노성훈(S . H . Noh),김호근(H . K . Kim),박준오(J . O . Park),이종인(J . I . Lee),유내춘(N . C . Yoo),김주항(J . H . Kim),노재경(J . K . Roh),김병수(B . S . Kim),강진경(J . K . Kang),민진식(J . S 대한내과학회 1997 대한내과학회지 Vol.52 No.1

Objectives : Prognosis of gastric cancer is related to invasion and metastasis. Evidence has accumulated that invasion and metastasis in solid tumors require the action of tumor associated proteases, which promote the dissolution of the surrounding tumor matrix and the basement membrane. The serine protease [(urokinase-type plasminogen activator and plasminogen activator inhibitor-1 (PAI-1)] and type IV collagenase(MMP-9 and MMP- 2) appear to play a key role in these processes. Recent reports have demonstrated that expression of these proteolytic enzymes are elevated in breast and colon cancer and that it can be associated with invasiveness and poor prognosis. We therefore evaluated whether the expression and activation of uPA, PAI-1 and type IV collagenase might be of clinical value in gastric cancer as a tumor/biologically defined risk factor. Methods: In a consecutive series of 160 gastric cancer patients who were enrolled in the Yonsei Cancer Center Study Group, the expression of uPA, PAI-1 and type IV collagenase was determined by ELISA, zymography and mmunohistochemical method. The results were as follows. Results: 1) Both uPA and PAI-1 levels were significantly higher in cancer tissues than no rmal (uPA; 9.4±8.7vs 5.3±3.1 ng/mg protein cytosol, PAI-1;10.9±9.1vs 5.8± 2.9 ng/mg protein cytosol), (p<0.001 respectively). Both high uPA and PAI-1 levels were associated with differentiation of the tumor(p=0.04, p=0.004), and a high PAI-1 level was associated with lymph nodes metastasis at an advanced stage (p=0.003, p=0.04). There was a correlation between the levels of uPA and PAI-1 expression in cancer tissues(r=0.57). 2) The activation ratio of MMP-9 and MMP-2 in cancer tissues 0.32±0.25, 0.27±0.34 were significantly higher than in normal tissue 0.19±0,27, 0.06± 0.16(p<0.001). The MMP-9 activation was associated with lymphnode metastasis and the MMP-2 activation was associated with distant metastasis(p=0.011, p=0.041). 3) In univariate analysis all of the proteolytic enzymes were associated with short relapse free survival, but in multivariate analysis only the high uPA expression was an independent prognostic parameter for short relapse free survival of the gastric cancer patients. Conclusion: These data indicate that uPA, PAI-1 and type IV collagenase were involved in the progression of gastric cancer at different points of time by different mechanisms. The combined expression and activation of these proteolytic enzymes were poor prognostic factors in gastric cancer patients, so new therapy based on these biologic behavior of the tumor in the same stage are clinically applicable. In particular, uPA is a new independent variable for the identification of patients at high risk, therefore therapy targetting uPA can be applied as a new treatment modality for gastric cancer.

면역성병인에 의한 혈관손상이 혈관내피세포의 혈액응고 조절기전에 미치는 영향에 관한 연구 ; Tissue - Type Plasminogen Activator , Plasminogen Activator Inhibitor - 1 및 von Willebrand Factor 의 변동에 관하여

박선양(Seon Yang Park),송영욱(Yeong Wook Song) 대한내과학회 1991 대한내과학회지 Vol.40 No.2

To elucidate the pathophysiologic mechanisms for hemorrhagic or thrombotic conditions induced by immune-mediated vascular injury, changes of the plasma levels of tissue-type plasminogen activator (t-PA) and type 1 plasminogen activator inhibitor (PAI-1), and von Willebrand factor ristocetin cofactor (vWf:RCo) activities were evaluated in seven patients with Henoch-Schonlein purpura (HSP) and 18 patients with systemic lupus erythematosus (SLE) with or without thrombotic complications. 1) Plasma PA activities and t-PA concentrations were not changed significantly in patients with HSP or SLE compared to those in normal controls. 2) Plasma PAI activities were increased significantly in HSP (57.7±5.1%, p<0.001) and SLE (51.6±8.6%, p=0.039) compared to those in normal controls (45.7±5.6%). In patients with SEL, thrombotic complications had no effect on the activities of plasma PAI-1. 3) PAI-1 concentrations were significantly elevated in patients with SLE (44.3±31.2AU/ml, p=0.015) compared to those in normal controls (23.4±8.8AU/ml). However, no difference in PAI-1 concentrations were found between the patients with or without thrombosis. PAI-1 concentrations were not significantly changed in HSP. 4) On fibrin autographic analysis of euglobulin fractions after SDS-PAGE, the intensity of 97K and 72K bands were increased. 5) vWf:RCo activities were greatly increased in patients with SLE (373.5±141.2%) with or without thrombosis. However, no significant difference was observed between the two groups. In conclusion, the increase of plasma PAI activities in patients with HSP appeared to be a compensatory mechanism for the hemorrhagic tendency characteristic of the disease, whereas, the increases of PAI activities, PAI-1 concentrations, t-PA-PAI-1 complexes and vWf:RCo activities might be a pathogenetic mechanism for the thrombotic conditions often complicated in patients with SLE.

폐경후 여성들에 있어 Fibrinogen, Plasminogen Activator Inhibitor Type 1(PAI-1), Lipoprotein (a) [LP(a)]와 심폐체력과의 관련성

제세영,박원하 대한스포츠의학회 1998 대한스포츠의학회지 Vol.16 No.2

The prevalence of CVD increases with advancing age in women, particularly after menopause. The risk is lower in physically active women relative to sedentary peers, but little is known regarding the relationship of physical fitness to plasma fibrinogen, plamingen activator inhibitor-1 (PAI-1), lipoprotein(a) [Lp(a)] in healthy postmenopausal women. The purpose of the study was to investigate the relation of cardiopulmonary fitness to plasma fibrinogen, PAI-1, lipoprotein(a), lipid and lipoprotein profile, fasting glucose, pressure in 316 healthy postmenopausal women(mena 54.2±4.8 years). VO2peak(ml/kg/min) was determined by graded exercise test based on mixing chamber analysis of expired gas. Three fitness categories were established on the basis of VO2peak. The result are revealed significant differences only in %BF, BMI across fitness categories. Although women in the high fitness group had favorable lipid rofiles, glucose, BP, PAI-1, and fibrinogen, they were not significantly different from the low or moderate fitness group. VO2peak was significantly correlated inversely with %BF(r=.29, p=0.0001), TC/HDL-C ratio(r=.12, p=0.0401), and positively with HDL-C(r=.13, p=0.0145), but not with fibrinogen and PAI-1(r=-.10, p=0.0639) in pearson correlation. Stepwise multiple regression indicated that %BF, fasting glucose and LDL-C were significantly associated with PAI-1(R^2=0.15), and fasting glucose, TC/HDL-C ratio and age were significantly associated with fibrinogen(R^2=0.08). In particular, there was no significant correlation of LP(a) with any other factors. In conclusion, although PAI-1 and fibrinogen are significantly associated with other risk factors such as %BF, fasting glucose, LDL-C and fasting glucose, TC/HDL-C ratio, age, respectively, Cardiopulmonary fintness does not influence significantly on the level of fibrinogen. But PAI-1 tend to decrease inversely with VO2peak in postmenopausal women.

욕창환자에서의 Plasminogen Activator Inhibitor-1 활성도의 변화

한창완,이진석,박운식,조영삼,이귀래,이홍복,변종훈,김미란,오도연 大韓血液學會 1996 大韓血液學會誌 Vol.31 No.1

Hyaluronic Acid Promotes Angiogenesis by Inducing RHAMM-TGF beta Receptor Interaction via CD44-PKC delta

박덕범,정두일,김영미,김현아,김경종,이윤실,최종선,한장희,이한수,전종욱,최철희,김영명 한국분자세포생물학회 2012 Molecules and cells Vol.33 No.6

Hyaluronic acid (HA) has been shown to promote angio-genesis. However, the mechanism behind this effect re-mains largely unknown. Therefore, in this study, the me-chanism of HA-induced angiogenesis was examined. CD44 and PKC were shown to be necessary for induction of the receptor for HA-mediated cell motility (RHAMM), a HA-binding protein. RHAMM was necessary for HA-pro-moted cellular invasion and endothelial cell tube formation. Cytokine arrays showed that HA induced the expression of plasminogen activator-inhibitor-1 (PAI), a downstream target of TGF receptor signaling. The induction of PAI-1 was dependent on CD44 and PKC. HA also induced an interaction between RHAMM and TGF receptor I, and induction of PAI-1 was dependent on RHAMM and TGF receptor I. Histone deacetylase 3 (HDAC3), which is de-creased by HA via rac1, reduced induction of plasminogen activator inhibitor-1 (PAI-1) by HA. ERK, which interacts with RHAMM, was necessary for induction of PAI-1 by HA. Snail, a downstream target of TGF signaling, was also necessary for induction of PAI-1. The down regulation of PAI-1 prevented HA from enhancing endothelial cell tube formation and from inducing expression of angiogenic factors, such as ICAM-1, VCAM-1 and MMP-2. HDAC3 also exerted reduced expression of MMP-2. In this study, we provide a novel mechanism of HA-promoted angiogenesis, which involved RHAMM-TGFRI signaling necessary for induc-tion of PAI-1.

뇌경색 환자에서 PAI-1 유전자의 다형성에 대한 연구

방차옥,안무영 순천향의학연구소 2000 Journal of Soonchunhyang Medical Science Vol.6 No.2

Background : Decreased fibrinolysis due to increased plasminogen activator inhibitior-1 (PAI-1) activity has been associated with hypertension or cerebral infarction. The aims of this study were to observe associations of the genetic polymorphism for PAI-1 with hypertension and cerebral infarction, and to elucidate whether impaired fibrinolytic activity in cerebral infarction was related to atherothrombosis or to risk factors such as hypertension. Methods : Patients with cerebral infarction (n=60), hypertension (n=100), and control subjects (m=100) were enrolled. We genotyped all subjects for 4G/5G polymorphism in the promoter region of the PAI-1 gene. Results : The frequency of 4G/4G genotype of PAI-1 was significantly higher in the patients of cerebral infarction than the control subjects (41.7% versus 21.0%; P=0.005), but not in the hypertensive subjects. There was a significant association between 4G/4G genotypes of PAI-1 and cerebral infarction (adjusted odds ratio=3.11, 95% confidence interval, 1.18-8.15), adjusting for age, sex, total cholesterol, low-density lipoprotein, triglyceride, and body mass index. Conclusion : Our results suggest that the 4G/4G genotype of the PAI-1 gene is significantly associated with an increased risk of cerebral infarction.

성인에서의 연령에 따른 내생적 플라스미노겐 활성제에 관한 연구

송인욱,하정호,정두신,어경윤 東國大學校醫學硏究所 2005 東國醫學 Vol.12 No.1

유로키나제형 플라스미노겐 활성제(Urokinase type plasminogen activator, u-PA)와 조직형 플라스미노겐 활성제(Tissue type plasminogen activator, 1-PA)는 현재 까지 알려져 있는 내생적 플라스미노겐 활성제로써 혈전용해력 뿐 만 아니라 세포이주, 조직파괴, 철관형성, 조직재건 등의 다양한 역할을 한다. 이런 내생적 플라스미노겐 활성제는 플라스미노겐 활성 억제제(Plasminogen activator inhibitor-1, PAI-1)에 의해서 강력하게 조절되고 있으나 사람에 있어서 이들 내생적 플라스비노겐 활성제들의 활성 정도가 연령에 의해 어떠한 영향을 주고 있는지는 아직 잘 알려져 있지 않다. 내생적 플라스비노겐 활성제의 연령에 따른 영향을 평가하기 위하여 전에 뇌경색의 과거력이 없는 성인 40명을 대상으로 chromogenic assay에 의해 u-PA, t-PA 그리고 PAI-1의 활성도를 측정하였다. t-PA 활성도는 40대에서는 100%, 50대에서는 90%, 60대에서는 30%, 70대에서는 30%였으며, u-PA의 활성도는 40대에서는 60%, 50대에서는 40%, 60대에서는 40%, 70대에서는 30%에서 나타났다. PAI-1 활성도는 40대에서는 10%, 50대에서는 30%, 60대에서는 40%, 70대에서는 70%에서 나타났다. 내생적 t-PA의 낮은 활성도와 PAI-1의 높은 활성도를 가진 고령에 있어서는 뇌에 만약 허혈 손상이 온다면 뇌의 섬유소 용해력이 떨어짐에 따라 결과적으로 크고 심한 뇌경색의 소인이 될 가능성이 충분히 있다고 하겠다. Urokinase-type plasminogen activator (u-PA) and tissue-type plasminogen activator (t-PA) play important roles in fibrinolysis, cell migration, tissue destruction, angiogenesis and tissue remodeling. u-PA and t-PA activity in tissue are tightly regulated by plasminogen activator inhibitor-1 (PAI-1). However, little is known of the effect of age on expression of endogenous plasminogen activators in human. To evaluate effect of age on endogenous plasminogen activators in normal human, we measured u-PA, t-PA and PAI-1 activity by chromogenic assay in 40 humans without previous ischemic stroke history. t-PA activity was present in 100% in 5th decade, 90% in 6th decade, 30% in 7th decade, and 30% in 8th decade health human. PAI-1 activity was present more increasing activity in 7th and 8th decade than 5th and 6th decade. Whereas, u-PA activity was not present significant difference. It is possible that lower t-PA activity and higher PAI-1 activity in older human may compromise brain fibrinolysis, predisposing to larger and more disabling cerebral infarction.

Propofol treatment modulates neurite extension regulated by immunologically challenged rat primary astrocytes: a possible role of PAI-1

고현명,주소현,이성훈,김희진,이승현,정재훈,류종훈,김정민,구본녀,신찬영 대한약학회 2015 Archives of Pharmacal Research Vol.38 No.4

Propofol, a widely used anesthetic, regulatesneurological processes including neurotoxicity, neuroprotection,glial activation, synaptic plasticity and neuronalmaturation. Tissue plasminogen activator/tissue plasminogenactivator inhibitor-1 (tPA/PAI-1) in CNS acts as aneuromodulator regulating synaptic plasticity, neuriteoutgrowth, seizure spreading and cell survival. Here, weinvestigated the effects of propofol on tPA/PAI-1 systemusing cultured neurons and astrocytes and their role in theregulation of neurite extension. Cultured rat primaryastrocytes were treated with propofol (1–10 lM) and LPS(10 ng/ml). The expression of functional tPA/PAI-1 wasexamined by casein zymography, Western blot and RTPCR. Alternatively, culture supernatants were added tocultured rat primary neuron to investigate the effects onneurite extension. Propofol alone did not affect tPA activityin rat primary cortical neuron. Similarly, propofol alonechanged neither tPA nor PAI-1 activity in rat primaryastrocytes. In immunologically challenged situation usingLPS, propofol synergistically increased expression of PAI-1 in rat primary astrocytes without affecting tPA expressionin a manner dependent on MAPKs activation. Increased expression of PAI-1 reduced tPA activity in LPSplus propofol-treated rat primary astrocytes. Consistentwith the critical role of tPA activity in the regulation ofneurite extension (Cho et al. 2013), the diminished tPAactivity in astrocyte culture supernatants resulted indecreased neurite extension when administered to culturedrat primary cortical neuron. The results from the presentstudy suggest that propofol, especially in immunologicallychallengedsituation, dysregulates tPA/PAI-1 system inbrain. Whether the dysregulated tPA/PAI-1 activityadversely affects neural differentiation as well as regenerationof neuron in vivo should be empirically determined inthe future.