Expression and function of Neutral Endopeptidase/CD10 on pancreatic adenocarcinoma Dissertation zur Erlangung des akademischen Grades doktor rerum naturalium (Dr. rer. nat.) vorgelegt der Naturwissenschaftlichen Fakultät I (Institut für Biologie) der Martin-Luther-Universität Halle-Wittenberg von Herrn Diplom Biologe Mabruk Erhuma H. Erhuma geboren am 19 August, 1965 Libyen Gutachter: 1. Prof. Dr. Hans-Jörg Ferenz 2. PD Dr. Hoang-Vu Cuong 3. Prof. Dr. Walter Halangk Halle (Saale), den 17.03.2008 urn:nbn:de:gbv:3-000013393[http://nbn-resolving.de/urn/resolver.pl?urn=nbn%3Ade%3Agbv%3A3-000013393] Thank God for all his blessings My family for continuous and unconditional support - III - Contents 1 INTRODUCTION 1 1.1 Pancreas anatomy, histology and function 1 1.2 Pancreas diseases 2 1.2.1 Pancreatitis 2 1.2.2 Pancreatic cancer 3 1.2.3 Risk factors 4 1.2.4 Cellular cancer classification of pancreatic adenocarcinoma 5 1.3 Ectoenzymes 6 1.3.1 Neutral endopeptidase 8 1.4 NEP/CD10 expression in pancreatitis and carcinoma tissues 14 1.5 Aim of the present investigation 15 2 MATERIALS AND METHODS 16 2.1 Pancreatic adenocarcinoma cell lines and culture conditions 16 2.2 Plasmids 16 2.3 RNA isolation and cDNA synthesis 18 2.4 NEP/CD10 standard preparation 18 2.5 Quantitative RT-PCR 19 2.6 Flow cytometry 20 2.7 Genomic DNA extraction 21 2.7.1 Bisulfite modification of DNA 21 2.7.