p53-mediated inhibition of angiogenesis in diffuse low-grade astrocytomas.
The p53 tumour suppressor protein has long been recognized as the central factor protecting humans from cancer. In this study we evaluated the associations of p53 status and vessel density (angiogenesis) in a set of diffuse low-grade astrocytomas. Immunohistochemistry was performed on 23 diffuse low-grade astrocytomas for CD31 and p53. Mutations in the TP53 gene were identified by PCR amplification of genomic DNA extracted from the indicated tumour tissues. Microvessel counts were done by computer analyses. Intratumoural or peritumoural microvascular hot spots were assessed and analysed from an image taken with a 200x fold magnification. Statistical analysis was performed with Pearson correlation coefficient and Student's t-test. We found that 9/23 (39%) of the astrocytomas stained positive for p53 in the immunohistochemistry. We identified TP53 mutations in 11/23 (47%) of the astrocytomas. No association between micro vessel density (MVD) and p53 immunohistochemical status was found. However, the MVD was significantly increased in p53 mutated low-grade astrocytomas. Furthermore, the absolute vessel number was significantly higher in p53 mutated than in p53 wild-type low-grade astrocytomas. To analyse the molecular background for that epiphenomenon LN229 glioma cells which harbour a TP53 mutation were transfected with a plasmid encoding p53 wild-type and an angiogenesis protein array was performed. We detected a significant increase for thrombospondin-1, coagulation factor III and serpin E1 and a significant decrease of MMP-9 in wild-type p53 transfected LN229 cells. The higher microvessel density and the increased absolute vessel number in p53 mutated tumours supports the importance of p53 for tumour angiogenesis in diffuse low-grade astrocytomas. Our results support the hypothesis that p53 regulates angiogenesis in low-grade astrocytomas.
Department of Neuropathology, University Hospital Heidelberg, Im Neuenheimer Feld 220, 69120 Heidelberg, Germany.
Timo
T
Gaiser
Maria R
MR
Becker
Jochen
J
Meyer
Antje
A
Habel
Markus D
MD
Siegelin
D001254:Astrocytoma / Q000473:pathology*; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000473:pathology*; D002196:Capillaries / Q000473:pathology; D045744:Cell Line, Tumor; D004252:DNA Mutational Analysis; D006801:Humans; D007150:Immunohistochemistry; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis / Q000235:genetics; D009389:Neovascularization, Pathologic / Q000473:pathology*; D040081:Protein Array Analysis; D040901:Proteomics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D016159:Tumor Suppressor Protein p53 / Q000502:physiology*
D016159:Tumor Suppressor Protein p53; D020780:Matrix Metalloproteinase 9