COMPARATIVE STUDY
JOURNAL ARTICLE

Apoptosis, necrosis and hypoxia inducible factor-1 in human head and neck squamous cell carcinoma cultures

Johann Gross, Julia Fuchs, Astrid Machulik, Volker Jahnke, Thomas Kietzmann, Ulrike Bockmühl
International Journal of Oncology 2005, 27 (3): 807-14
16077932
The objective of this study was to examine the mode of cell death and the hypoxia inducible factor-1 (HIF-1) expression of human head and neck squamous cell carcinoma (HNSCC) exposed to hypoxia in vitro. Apoptosis and necrosis rates were examined using flow cytometry. The findings suggest that HNSCC cells show a considerable heterogeneity in cell size and in response to hypoxia. A small-cell population showed a high spontaneous apoptosis and necrosis rate which was in-sensitive to hypoxia. A large-cell population responded to hypoxia by increase of apoptosis rate in parallel to recruitment of HIF-1. Hypoxia led to increased HIF-1alpha protein levels in nuclear extract using ELISA-binding activity. In all cells, accumulation of HIF-1 in the nuclei during hypoxia and a rapid degradation of HIF-1 in the post-hypoxic period were observed immunocytochemically. The HIF-1alpha mRNA level showed an expression of 10-40 pg/microg total RNA and remained unchanged in one cell line, while slightly decreasing in the other. Remarkably, no increased luciferase activity response was found on the reporter gene level using pGL3 reporter gene with three erythropoietin hypoxia responsive elements, either by hypoxia or by application of lactacystin, desferrioxamine or CoCl2. These findings suggest that, in HNSCC cells, hypoxia induces HIF-1alpha to stabilize and accumulate in the cell nuclei but have a cell-specific transcriptional complex.

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