Journal Article

PHD3 regulates differentiation, tumour growth and angiogenesis in pancreatic cancer

Görlach, A, Friess, H, Hines, O J, Su, Y, Loos, M, Diebold, I, Metzen, E, Büchler, P, Giese, N, Pastorekova, S

2010

Overview

Purpose: Tumour hypoxia activates hypoxia-inducible factor-1 (HIF-1) and indluences angiogenesis, cell survival and invasion. Prolyl hydroxylase-3 (PHD3) regulates degradation of HIF-1 α . The effects of PHD3 in tumour growth are largely unknown. Experimental design: PHD3 expression was analysed in human pancreatic cancer tissues and cancer cell lines by real-time quantitative PCR and immunohistochemistry. PHD3 overexpression was established by stable transfection and downregulation by short interfering RNA technology. VEGF was quantified by enzyme-linked immunosorbent assay. Matrigel invasion assays were performed to examine tumour cell invasion. Apoptosis was measured by annexin-V staining and caspase-3 assays. The effect of PHD3 on tumour growth in vivo was evaluated in an established orthotopic murine model. Results: PHD3 was upregulated in well-differentiated human tumours and cell lines, and regulated hypoxic VEGF secretion. PHD3 overexpression mediated tumour cell growth and invasion by induction of apoptosis in a nerve growth factor-dependent manner by the activation of caspase-3 and phosphorylation of focal adhesion kinase HIF-1 independently. In vivo , PHD3 inhibited tumour growth by abrogation of tumour angiogenesis. Conclusion: Our results indicate essential functions of PHD3 in tumour growth, apoptosis and angiogenesis and through HIF-1-dependent and HIF-1-independent pathways.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/208/200 / 692/699/67/1504/1713 / Adenocarcinoma - pathology / Adenocarcinoma - surgery / Animals / Annexin A5 - analysis / Apoptosis / Biological and medical sciences / Biomedical and Life Sciences / Biomedicine / Cancer Research / Carcinoma, Pancreatic Ductal - enzymology / Carcinoma, Pancreatic Ductal - pathology / Carcinoma, Pancreatic Ductal - surgery / Caspase 3 - metabolism / Cell Differentiation / Cell Line, Tumor / Dioxygenases - genetics / Dioxygenases - physiology / Drug Resistance / Enzyme-Linked Immunosorbent Assay / Epidemiology / Gastroenterology. Liver. Pancreas. Abdomen / Humans / Hypoxia-Inducible Factor 1, alpha Subunit - genetics / Hypoxia-Inducible Factor 1, alpha Subunit - physiology / Hypoxia-Inducible Factor-Proline Dioxygenases / Liver. Biliary tract. Portal circulation. Exocrine pancreas / Medical sciences / Mice / Molecular Medicine / Neoplasm Invasiveness - genetics / Neoplasm Invasiveness - pathology / Neovascularization, Pathologic - pathology / Oncology / Pancreatic cancer / Pancreatic Neoplasms - enzymology / Pancreatic Neoplasms - pathology / Pancreatic Neoplasms - surgery / Polymerase Chain Reaction / Reverse Transcriptase Polymerase Chain Reaction / RNA, Messenger - genetics / RNA, Neoplasm - genetics / Translational Therapeutics / Transplantation, Heterologous / Tumors / Up-Regulation / Vascular Endothelial Growth Factor A - analysis