Strong inhibition of NF-kB activity was found in extracts of leaf and rhizome from Nuphar lutea L. SM. (Nuphar). The inhibitory action was narrowed down to a mixture of thionupharidines and/or thionuphlutidines that were identified in chromatography fractions by one- and

two-dimensional NMR analysis. Dimeric sesquiterpene thioalkaloids were identified as the major components of the mixture. The Nuphar alkaloids AUY-922 chemical structure mixture (NUP) showed a dose dependent inhibition of NF-kB activity in a luciferase reporter gene assay as well as reduction of nuclear NF-kB subunits expression as tested by western blots and immunohistochemistry. Decreased DNA binding was demonstrated in Electro Mobility Shift Assays (EMSA). NUP

inhibited both inducible and constitutive NF-kB activation and affected the canonical and alternative pathways. Tideglusib in vitro Suppression of NF-kB was not cell type specific. Induction of apoptosis by the alkaloid mixture was demonstrated by time-dependent and dose-dependent cleavage of procaspase-9 and PARP. Synergistic cytotoxicity of the active mixture with cisplatin and etoposide was demonstrated. In addition, NUP partially protected mice from LPS- induced septic shock and from experimental B16 melanoma lung metastasis. Overall, our results show that NUP inhibits the NF-kB pathway and acts as a sensitizer to conventional chemotherapy, enabling the search for its specific target and its application against cancer and inflammation. Poster No. 46 Molecular Dissection of the Pro-metastatic Effects of ASAP1 Anna Poletti 1 , Thomas Müller2, Ulrike Stein3, Nicoletta Tata4, Livia Garzia4, Massimo Zollo4, Jonathan Sleeman1,2 1 Department of Microvascular Biology, Universitätsmedizin Mannheim, University of Heidelberg, Mannheim, Germany, 2 Department of Toxicology and Genetics, Forschungszentrum

Karlsruhe, Karlsruhe, Germany, 3 Department of Surgery and Surgical Oncology, Gene Therapy Group, Max-Delbrück-Center for Molecular Medicine, PIK3C2G Charité-Universitätsmedizin Berlin, Berlin, Germany, 4 CEINGE, Centro di Ingegneria Genetica e Biotecnologia, Naples, Italy To understand the molecular mechanisms that underlie the metastatic process is of pivotal selleckchem importance in cancer research. In an unbiased genetic screen for genes that are involved in metastasis formation we identified ASAP1 (Arf-GAP with SH3-domains, Ankyrin-repeats and PH-domains), and subsequently showed that it promotes tumor cell motility and invasiveness. Loss and gain of function experiments in a pancreatic carcinoma model demonstrate a functional role for ASAP1 in regulating metastasis. In human colorectal cancer patients we found that ASAP1 expression strongly correlates with short metastasis-free survival and poor prognosis.