Oncology reports, 2008-08, Vol.20 (2), p.413-419
2008
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- Autor(en) / Beteiligte
- Titel
- Rapid detection of the hypoxia-regulated CA-IX and NDRG1 gene expression in different glioblastoma cells in vitro
- Ist Teil von
- Oncology reports, 2008-08, Vol.20 (2), p.413-419
- Ort / Verlag
- Athens: S.n.
- Erscheinungsjahr
- 2008
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Hypoxia-inducible factor-1 (HIF-1) is a key regulator of tumor cell hypoxia. It regulates the expression of several genes related to oxygen homeostasis in response to hypoxic stress. Carbonic anhydrase IX (Ca-IX) has been found to be a stable marker of acute or chronic hypoxia. N-Myc down-regulated gene 1 (NDRG1) has been shown to possess more specific characteristics for clinical analysis and identification purposes. HIF-1 activates gene expression of the two genes and promotes tumor cell survival under hypoxic conditions. Herein, we modified a flow cytometry protocol to separate NDRG1- and CA-IX-negative and -positive cells in vitro to sort chronically hypoxic cells from glioblastoma tumors. The FITC-anti-CA-IX fluorescence differed between positive and negative cells by a factor of 60-160 in U373, U87-MG, U251 and GaMG, respectively. A clear effect of the O2 concentration on CA-IX expression was visible in GaMG and U251 cell lines whereas U373 showed a less differentiated pattern. NDRG1 expression was present in U373, U251 and GaMG with the lowest expression rate in GaMG. It was stable over 48 h of reoxygenation after 24 h of extreme hypoxia (0.1% O2). During reoxygenation NDRG1 was relatively stable in the four tumor cell lines with the lowest expression in GaMG. An oxygen- and time-dependent elevation of nuclear HIF-1alpha binding on HRE was displayed. FACS analysis of CA-IX and NDRG1 expression may be a new approach to determining the hypoxic state of tumor cells. However, an extensive analysis of other hypoxia-regulated genes in different tumors is required to identify additional markers for the detection of the oxygenation state in human tumors in order to tailor effective tumor-specific therapeutic strategies.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1021-335XeISSN: 1791-2431DOI: 10.3892/or_00000023Titel-ID: cdi_proquest_miscellaneous_69319711
- Format
- –
- Schlagworte
- Antigens, Neoplasm - genetics, Antigens, Neoplasm - metabolism, Biological and medical sciences, Blotting, Western, Brain Neoplasms - genetics, Brain Neoplasms - pathology, Carbonic Anhydrase IX, Carbonic Anhydrases - genetics, Carbonic Anhydrases - metabolism, Cell Cycle Proteins - genetics, Cell Cycle Proteins - metabolism, Cell Nucleus - metabolism, Gene Expression Regulation, Neoplastic, Glioblastoma - genetics, Glioblastoma - pathology, Humans, Hypoxia - genetics, Hypoxia-Inducible Factor 1, alpha Subunit - metabolism, In Vitro Techniques, Intracellular Signaling Peptides and Proteins - genetics, Intracellular Signaling Peptides and Proteins - metabolism, Medical sciences, Neurology, Oxygen - metabolism, Response Elements - genetics, RNA, Messenger - genetics, RNA, Messenger - metabolism, Tumor Cells, Cultured, Tumors, Tumors of the nervous system. Phacomatoses