[C'ANC'ER K|-:S|-:AKC|| SS. .«SU-MM. September 15, l")95| Advances in Brief Primary Tumor and Metastasis in Ovarian Cancer Differ in Their Content of Urokinase-type Plasminogen Activator, Its Receptor, and Inhibitors Types 1 and 21 B. Schmalfeldt,- W. Kühn,U. Reuning, L. Pache, P. Dettmar, M. Se-limiti, F. Jänicke, H. Höfler,and H. Graeff Frauenklinik der Technischen Universität München. Klinikum rechts tier Isar. Ismaninger Strasse 22. D-KI675. Munich ¡B.S.. W. K., U. R., L P.. M. S., F. J., H. G.I. and Institut fürAllgemeine Pathologie und Pathologische Anatomie, Technische Universität München¡P.D., H. H. I, Munich, Germany Abstract metastasis is controlled by a series of successive events. After local proteolysis of the surrounding extracellular matrix, detachment and The relevance of urokinase-type plasminoceli activator (uPA) and plas- spread of tumor cells is facilitated followed by the invasion of the minogen activator inhibitor (PAI) type I in predicting the survival prob adjacent tissue, crossing of tissue boundaries and the basement mem ability of patients with advanced ovarian cancer after radical surgery and adjuvant chemotherapy by assessing the patients' primary tumors has brane. Intravasation and extravasation are followed by reimplantation recently been shown by us (W. Kühnelal., Gynecol. Oncol., 55: 401-409, of tumor cells into the respective target organ, remodeling of a new 1994). In the present study, we determined uPA, uPA receptor, PAI-1, and tumor stroma, and angiogenesis in order to consolidate a secondary PAI-2 concentrations in primary tumors and tumor-infiltrated omentum tumor at a distant locus. For these events a fine-tuned balance between and rctroperitoneal lymph nodes of ovarian cancer patients. The group proteolytic factors, their inhibitors, adhesive proteins, angiogenic fac consisted of 39 patients with advanced ovarian carcinoma stages Fédéra tion Internationale de Gynécologieet d'Obstétriqueil-ÕGOiHic or IV; for tors, oncoproteins, and growth factors has to be adjusted (2, 3). Basic and clinical research in tumor invasion and metastasis have comparison 7 patients with early carcinoma stage FIGO I were also focused on the role of tumor-associated proteases such as metallopro- included. In métastasesof the omentum from ovarian cancer stage FIGO lile or IV patients, we noted a 4-fold elevated uPA content, a 2-fold teinases and the serine proteases of the plasmin/plasminogen activator increase in PAI-I, and also a significant increase in uPA receptor and system (2). Metalloproteinases seem to be correlated with tumor cell PAI-2 over primary tumors. In métastasesof the lymph nodes the levels of invasion and metastasis in animal models; however, conclusive data the respective antigens were also increased when compared to primary on their clinical significance in humans are still lacking. In contrast, tumors. These data may indicate that elevated levels of components of the for several different solid cancers a strong prognostic impact of the fihrinolytic system at sites of métastasesmay contribute to the aggressive scrine protease uPA1 and/or its inhibitor PAI-1 as predictors for the potential of cancer cells by favoring their rcimplantation and/or the course and the outcome of the cancer has been reported. Besides their consolidation of a new tumor stroma. statistically independent prognostic impact in cancer of the breast (4, Introduction 5), lung (6), colon (7), and the gastrointestinal tract (8), significantly elevated levels of both uPA and PAI-1 have been demonstrated in Ovarian carcinoma is one of the most severe gynecological malig ovarian cancer tissues compared to those in normal ovarian tissue nancies and because of a long asymptomatic course of the disease specimens. Patients with advanced ovarian cancer can be subgrouped displays already advanced stages at diagnosis with extensive cancer on the basis of computer-optimized "cutoff values for uPA and spread. This is reflected by the high incidence (70% of the cases) of PAI-1: patients with low uPA and PAI-1 (uPA < 0.9 ng/mg protein; mortality within 5 years, a fact that stresses the necessity of an early PAI-1 < 13.5 ng/mg protein) had a statistically better prognosis than diagnosis for risk selection and the prediction of the course of the patients with high uPA and PAI-1 antigen levels (9). First indications disease. Several investigators have demonstrated that the prognosis of for a role of the uPA-mediated proteolytic system in tumor invasion patients with advanced ovarian cancer could be improved by radical and metastasis came in 1976 from Astedt and Holmberg (10), who surgery followed by adjuvant chemotherapy. Although the absence of demonstrated for the first time high uPA concentrations in cultured residual tumor after cytoreductive surgery proved to be a strong ovarian carcinoma tissue. In the following years increased uPA and prognostic factor for survival, the complete resection of the tumor PAI-1 levels were also found in tumor tissue extracts of patients with mass was not sufficient to protect all patients from the recurrence of ovarian carcinoma (11, 12) and in malignant ascites (13). the disease and subsequent death. Several attempts have been under uPA is synthesized and secreted by normal and tumor cells. Both taken to improve the prognosis of cancer patients and to establish the inactive proenzyme form of uPA, pro-uPA, and protcolytically predictive factors that are related to tumor biology. Numerous factors active uPA bind to a specific glycanlipid-anchored receptor (uPA-R, have been screened, among them the receptors for estrogen, proges CD 87) on the tumor cell surface. Upon binding of pro-uPA, its terone, and epidermal growth factors, DNA ploidy, S-phase, and activation by other proteases (e.g., plasmin) is facilitated. uPA acti oncogenes ( 1). In recent years special attention has been paid to tumor cell surface- vates the proenzyme plasminogcn into the broad spectrum serine protease plasmin, which may in turn either directly degrade extracel associated proteolytic enzyme systems regarding their impact on lular matrix proteins such as fibrin, fibronectin, laminin, and proteo- cancer progression and metastasis. The process of tumor invasion and glycans or activates certain matrix-degrading enzymes such as pro- collagenase. The activation of uPA is controlled by its specific- Received ft/15/95; accepted 8/4/95. inhibitors PAI-1 and PAI-2. Binding of PAI-1 or PAI-2 to uPA-R- The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore he hereby marked advertisement in accordance with bound uPA results in the subsequent internalization of the ternary 18 U.S.C'. Section 1734 solely to indicate this fact. 1This work was supported by the Deutsche Forschungsgemeinschaft (Klinische For schergruppe GR 280/4-1) and the BIOMF.D I program (BMH1CTO1346). 1The abbreviations used are: uPA. urokinase-type plasminogcn activator; PAI, plas- : To whom requests for reprints should be addressed. minogen activator inhibitor; uPA-R. uPA receptor. 3958 Downloaded from cancerres.aacrjournals.org on September 30, 2021. © 1995 American Association for Cancer Research. iil'A. iil'A-R. AND PAI-1/2 IN PRIMARY TUMOR AND MUTASI ASÕS complex, thereby regulating cell surface plasmin generation. uPA-R minorversusTable 2 Owi/ximwi ofiiPA. uPA-R. PAI-1.anil PAI-2 utilicen /nr/.v//( primary IVIAntigen(ng/mgtuimir-injiliratal iimenlum nuijus in iul\tinced tmirian cunïmmii ír'líiOlile ¡ir in combination with specific plasmin(ogen) receptors focalizes the FIGOIlle/IVtumor metastasis(« plasmin/plasminogen activator system to the tumor cell surface, thus (11=39)"(UM(0.05-10.16)2.87(0.55-8.66)16.99(0.14-105.1)0.25'(0.01-11.34)Omentum=33)"3.65*(0.66-16.58)4.3o(1.15-8.71)34.14(4.60-125.3)1.72(0.01-12.39)Statisticalsignificance protein)uPA(median (P)II.IKIOI0.050.00090.1104 constituting an effective proteolytic enzyme system (2). Tumor-associated proteolytic enzyme systems are not only impor range)uPA-R(median tant for tumor cell spread from the primary tumor but seem to be crucial parameters for the events at the site of métastases.In this report we investigated the levels of uPA, uPA-R, PAI-1, and PAI-2 in range)PAI-1(median tissue extracts of primary tumors, tumor-infiltrated omentum, and tumor-infiltrated lymph nodes of patients suffering from advanced range)PAI-2(median ovarian carcinoma. range)Primary Patients and Methods " il. number of patients. Forty-six patients with early (FIGO I) or advanced ovarian cancer stages '' n = 34. ' n = 38. (FIGO lile or IV) were enrolled as par! of a prospective study on ovarian cancer patients undertaken at the Frauenklinik der Technischen Universität München(Table 1). Patients with stages FIGO II, Ilia, and Illh were excluded from the present analysis because of a too small number of eases. Patients with assisted analysis (EIA program; ICN-Flow Laboratories, Meckenheim. Ger advanced ovarian carcinoma received the following radical surgical treatment: many). Protein concentrations were determined using the BCA Protein Assay longitudinal laparotomy with hysterectomy, bilateral adenectomy. appendec Reagent kit (Pierce, Rockford, IL). Values for uPA, uPA-R, PAI-1, and PAI-2 tomy, infragastric omentectomy, and bilateral pelvic and paraaortic lymphad- concentrations are expressed as ng/mg protein of the tissue extracts. In our enectomy. Among the group of patients, seven patients received limited laboratory the interassay variation coefficient for uPA was 5.8%, for uPA-R surgical treatment because of their had health condition. In younger patients 5.6%, for PAI-1 4.0%, and for PAI-2 1.4%, thus fulfilling the criteria for a (<35 years) with tumor stage FIGO I.