gms | German Medical Science

27th German Cancer Congress Berlin 2006

German Cancer Society (Frankfurt/M.)

22. - 26.03.2006, Berlin

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cDNA Microarray analysis of apoptotic effects of different chemotherapeutics in human fibrosarcoma cells

Meeting Abstract

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  • corresponding author presenting/speaker Adrien Daigeler - Universitätsklinik für Plastische Chirurgie und Schwerbrandverletzte, Handchirurgiezentrum, Operatives Referenzzentrum für Gliedmaßentumoren, Bochum, Deutschland
  • Cornelius Kuhnen - Institut für Pathologie der Ruhr-Universität, Bochum
  • Oliver Müller - Tumorgenetik des Max-Planck-Intituts für molekulare Physiologie, Dortmund
  • Ludger Klein-Hitpass - Institut für Zellbiologie der Universität, Duisburg-Essen
  • Hans-Ulrich Steinau - Universitätsklinik für Plastische Chirurgie und Schwerbrandverletzte, Handchirurgiezentrum, Operatives Referenzzentrum für Gliedmaßentumoren, Bochum
  • Marcus Lehnhardt - Universitätsklinik für Plastische Chirurgie und Schwerbrandverletzte, Handchirurgiezentrum, Operatives Referenzzentrum für Gliedmaßentumoren, Bochum

27. Deutscher Krebskongress. Berlin, 22.-26.03.2006. Düsseldorf, Köln: German Medical Science; 2006. DocPO493

The electronic version of this article is the complete one and can be found online at: http://www.egms.de/en/meetings/dkk2006/06dkk603.shtml

Published: March 20, 2006

© 2006 Daigeler et al.
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Outline

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Introduction: Soft tissue sarcomas are notoriously resistant against most chemotherapeutic agents and therefore chemotherapy is not clinically established. The goal of this study was to elucidate the mechanisms and degree of response of a fibrosarcoma cell line after treatment with clinical used cytostatics.

Material and Methods: HT1080 fibrosarcoma cells were exposed with the one of the following chemotherapeutics: 1.) doxorubicin 2.) actinomycin d 3.) vincristine or carrier control. Total RNA was isolated and cDNA microarray (Affymetrix, 14.500 genes) was performed after either 6 or 24 h post treatment. A combination of three stepwise statistical approaches resulted in 3.309 (actinomycin d), 1.019 (doxorubicin) and 134 (vincristine) highly significant probe sets that clearly were altered expressed. Expression levels for 90 selected candidate genes were validated by quantitative real-time PCR resulting in a 93% agreement between the two techniques.

Results: For the RNA blocker actinomycin 99% of the genes were downregulated, in contrast to doxorubicin where 65% of the genes were upregulated. Our results indicate that doxorubicin mediates apoptosis by inducing the mitochondrial- (intrinsic) pathway. Despite the fact that p53 was unchangend we found increased levels of cytochrome c, APAF-1 and different member of the STAT-family (STAT1+3). In contrast Bcl-2 was decreased. Different caspases, including Casp 1, -3, -6, -8 and Casp-9, were increased indicating these molecules as key factors in doxorubicin mediated apoptosis.

Conclusion: This study demonstrates that fibrosarcoma cells reveal a specific response pattern on mRNA level, which is dependent of the cytostatic used. We hypothesize that ex vivo testing of cytostatics after tumor resection identifies a signature profile, which might improve help in decision making whether chemotherapy for the heterogeneous group of soft tissue sarcomas is beneficial or not. The doxorubicin mediated apoptosis pathways help understanding the diverse modes of soft tissue sarcoma cell death following cytotoxic therapies and could provide a molecular basis for new strategies.