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Gene expression of neuronal nitric oxide synthase and adrenomedullin in human neuroblastoma using real‐time PCR
Author(s) -
Dötsch Jörg,
Harmjanz Arndt,
Christiansen Holger,
Hänze Jörg,
Lampert Fritz,
Rascher Wolfgang
Publication year - 2000
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/1097-0215(20001015)88:2<172::aid-ijc4>3.0.co;2-i
Subject(s) - adrenomedullin , nitric oxide synthase , neuroblastoma , gene expression , gene , nitric oxide , microbiology and biotechnology , real time polymerase chain reaction , chemistry , neuronal nitric oxide synthase , messenger rna , biology , cancer research , biochemistry , endocrinology , genetics , cell culture , peptide
The objective of our study was to assess the gene expression of the antiproliferative systems neuronal nitric oxide synthase (nNOS) and adrenomedullin (AM) in human neuroblastoma. A novel real‐time PCR method was evaluated using neuropeptide Y (NPY) for validation. Glyceraldehyd‐3‐phospate dehydrogenase (GAPDH) and NPY gene expression in neuroblastomas of 50 patients were measured in parallel by competitive quantitative and TaqMan real‐time RT‐PCR. AM and nNOS mRNA were determined by real‐time PCR. Our results showed a linear relationship between competitive quantitative and real‐time RT‐PCR measurements of NPY and GAPDH (r = 0.87 and r = 0.92, respectively). AM and nNOS mRNA was found in all tumor samples. AM/GAPDH mRNA increased with higher differentiation according to Shimada ( p = 0.013). There was no relation between MYCN amplification nor with the tumor stage ( p = 0.78 and p = 0.30, respectively). AM/GAPDH did not relate to recurrence or death in a 5‐year follow‐up period. Neuronal NOS/GAPDH expression did not relate to any biological or clinical parameter of prognosis or differentiation. Similar results were obtained when the neuronal marker protein gene product 9.5 (PGP9.5) was used to normalize mRNA concentration. In conclusion, TaqMan real‐time PCR appears to be a reliable method to quantify gene expression in neuroblastomas. Adrenomedullin mRNA in neuroblastoma is linked to tumor differentiation but not to prognostic markers. Int. J. Cancer 88:172–175, 2000. © 2000 Wiley‐Liss, Inc.