Elevated Expression Level of Survivin Protein in Soft-Tissue Sarcomas Is a Strong Independent Predictor of Survival 1 Matthias Kappler, 2 Matthias Kotzsch, 2 Frank Bartel, Susanne Fu ¨ ssel, Christine Lautenschla ¨ger, Uta Schmidt, Peter Wu ¨ rl, Matthias Bache, Hannelore Schmidt, Helge Taubert, 3 and Axel Meye Institute of Pathology [M. Ka., F. B., H. S., H. T.], Institute of Medical Biometry and Informatics [C. L.], and Department of Radiotherapy [M. B.], University Halle-Wittenberg, D-06097 Halle/ Saale, Germany; Institute of Pathology [M. Ko.], and Department of Urology [S. F., U. S., A. M.], Technical University Dresden, Dresden, Germany; and Clinic of General and Transplantation Surgery [P. W.], University Ulm, Ulm, Germany ABSTRACT Purpose: Survivin is a member of the inhibitor-of- apoptosis gene family and is known to be overexpressed in a number of tumor types. The aim of this study was to eval- uate the prognostic value of survivin protein expression in tumor tissue extracts in a group of well-characterized soft- tissue sarcoma (STS) patients. Experimental Design: In this investigation, malignant tissue samples from 63 STS patients as well as from a panel of tumor cell lines were investigated, with nonmalignant tissues serving as controls. The survivin protein level was quantified by a novel ELISA and by Western blot analysis. Results obtained by both methods were compared with clin- icopathological parameters regarding tumor grade and tu- mor entity, and they were then correlated to survival in a multivariate Cox regression model. Results: High survivin levels were detected by ELISA and Western blot analysis in tumor tissue extracts and in lysates of tumor cell lines. None or only weak expression of survivin protein was found in nonmalignant cells and tis- sues. When comparing survivin values obtained by ELISA or Western blot, we found a significant correlation between both methods (P  0.013, Pearson test). Our findings re- vealed that, in multivariate Cox regression analyses, sur- vivin levels measured by ELISA and Western blot were significantly associated with tumor-related death in STS patients (P  0.001, RR  19.8, and P  0.004, RR  5.1, respectively). However, in a direct comparison of both sur- vivin protein detection assays, we found a higher sensitivity and a stronger correlation to prognosis in survivin ELISA as compared with the Western blot assays. Furthermore, a higher tumor grade and more aggressive STS entity showed an elevated survivin protein expression level. Conclusion: Altogether, an elevated survivin content in tumor tissue extracts has a significant and independent negative predictive value on the survival-rate of STS pa- tients. This finding corresponds well to data obtained for the mRNA level of survivin, as shown previously (M. Kappler et al., Int. J. Cancer, 95: 360 –363, 2001). INTRODUCTION Survivin, a member of the inhibitor-of-apoptosis protein family, represents a multifunctional protein that suppresses ap- optosis and regulates cell division at the G 2 -M phase (reviewed in Refs. 1 and 2). It is a nuclear shuttle protein that is actively exported from the nucleus (3). Independently, survivin expres- sion is also correlated to mitotic activity (4 – 6). Survivin is strongly overexpressed in a vast majority of cancers, and it is one of the most tumor-specific human gene products (7). The potential utility of survivin overexpression in early diagnosis and as a prognostic marker of cancer is incontrovertible (1). A correlation between survivin detection and the outcome of the affected tumor patients was described for different carcinoma types including colorectal cancer (8 –11), bladder cancer (12), lung carcinoma (13), breast cancer (14), esophageal cancer (15), pancreatic duct cell cancer (16), hepatocellular cancer (17), gastric cancer (18), and malignant glioma (19). Survivin seems to have strong potential as a tumor marker for malignant solid tumors of mesodermal origin when corre- lated to tumor progression and prognosis. In addition to the studies performed on neuroblastoma by Adida et al. and others (20 –23) we found that a high survivin transcript level signifi- cantly correlates with a poor outcome in adult STS 4 patients (24). More recently, we have shown a significant correlation between the coexpression of the survivin mRNA and the telom- erase reverse transcriptase mRNA and a highly increased risk of tumor-related death in the same group of STS patients (25). Moreover, several reports describe therapeutic implications of the inhibition of survivin expression for different malignan- Received 8/5/02; revised 11/18/02; accepted 11/19/02. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 1 Supported by Grant 3347A/0021B from the Ministry of Science and Education of the State Saxony-Anhalt and in part by Grant 99.009.1 from the Wilhelm Sander-Stiftung. 2 M. Ka., and M. Ko. contributed equally to the results of the study. 3 To whom requests for reprints should be addressed, at Institute of Pathology, Faculty of Medicine, University Halle-Wittenberg, Magde- burger Strasse 14, D-06097 Halle/Saale, Germany. Phone: 49-345-557- 1293; Fax: 49-345-557-1295; E-mail: helge.taubert@medizin.uni- halle.de. 4 The abbreviations used are: STS, soft-tissue sarcoma; Ab, antibody; RR, relative risk; WB, Western blot; RMS, rhabdomyosarcoma; LMS, leiomyosarcoma; MFH, malignant fibrous histiocytoma; FS, fibrosarcoma. 1098 Vol. 9, 1098 –1104, March 2003 Clinical Cancer Research Cancer Research. on February 16, 2016. © 2003 American Association for clincancerres.aacrjournals.org Downloaded from