Molecular radiobiologyExpression patterns of the hypoxia-related genes osteopontin, CA9, erythropoietin, VEGF and HIF-1α in human glioma in vitro and in vivo☆
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Cell and culture and hypoxia treatment
Early-passage human malignant glioma cell lines U251 and U373 from the American Type Culture Collection (ATCC, Rockville, MD) and GaMG, a cell line that was established from a patient with glioblastoma multiforme (Gade Institut of the University Bergen, Norway) [1], were grown on glass Petri dishes in Dulbecco’s modified Eagle’s medium, supplemented with 10% fetal bovine serum (FBS) and non-essential amino acids. Additionally, all culture media were supplemented with penicillin (100
Regulation of hypoxia-inducible gene expression at the mRNA level in vitro in human GBM cell lines and in vivo in human GBM and LGA tumor tissue
For all genes investigated, the time course of mRNA expression in human GBM cell lines in response to hypoxic exposure in vitro at O2 concentrations of 5%, 1% and 0.1% is displayed in Fig. 1. OPN displayed a strong hypoxic induction with increasing duration and severity of hypoxia in all three cell lines (Fig. 1a). CA9 was overexpressed under hypoxia in all three lines with the highest in GaMG (3-fold) (Fig. 2a). EPO was overexpressed only at moderate hypoxia (1% O2), but not at 0.1%, and VEGF
Discussion
Due to the poor prognosis of patients with glioblastoma multiforme, intensified and individualized treatment approaches are needed to improve the results of therapy. The presence of tumor hypoxia is an adverse prognostic factor, but may represent an opportunity to specifically target hypoxic tumor cells. Needle electrode studies have shown that oxygenation in GBM is lower than in surrounding brain tissue [5] and in immunohistochemical studies, perinecrotic regions of GBM stained positive for
Conflict of interest statement
The authors declare that they have no conflict of interest related to the contents of this manuscript.
Acknowledgements
The authors thank Thomas Freiman (Department of Neurosurgery, University of Freiburg, Germany) for normal brain control tissue, Bayer Healthcare Co., for provision of the M75 monoclonal antibody and Stefanie Gerngras and Astrid Katzer for technical assistance. This work was supported in part by a grant from the Deutsche Forschungsgemeinschaft (VO 871/2) to D.V. and by IZKF Würzburg (B25) to C.H. and G.H.V.
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2018, Journal of Oral and Maxillofacial Surgery, Medicine, and PathologyCitation Excerpt :The effect of CA IX on the tumor microenvironment is characterized by the regulation of pH [18]. Recently, some human tumors have been reported to express CA IX, which is proposed to influence the clinical features of these tumors [4,5,7–9,16]. Thus, HIF-1α and CA IX play a role in angiogenesis, apoptosis, invasion, and metastasis of malignant tumors [8,9].
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The author responsible for distribution of materials integral to the findings presented in this article is Dr. Harun M. Said.
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These two authors contributed equally.