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Cell cycle regulation of NF-kappa b-binding activity in cells from human glioblastomas.

Glioblastoma multiforme is a highly malignant and anaplastic tumor of the central nervous system representing more than 50% of all malignant gliomas. The cell origin of this highly undifferentiated tumor remains obscure, although it is postulated that glioblastomas are developed from astrocytes. The rapid growth of the glioma and the state of its undifferentiation are attributed to the deregulation of several signal transduction pathways and cell cycle events. Recent studies showed diverse functions for the NF-kappa B/Rel family of inducible transcription factors including differentiation, apoptosis, oncogenesis, and cell cycle regulation. We sought to examine the level of NF-kappa B activity throughout the glioma's cell cycle. Results from band-shift studies indicated a biphasic NF-kappa B DNA-binding activity in the nuclei of cycling glioblastoma cells. We showed that NF-kappa B-binding activity maximizes in nuclear extracts at specific cell cycle stages including G0/G1, mid-late G1, and S phase. Results from Northern blotting studies revealed that the differential expression of the NF-kappa B subunits, p50 and p65, may not be responsible for cell cycle stage-specific association of NF-kappa B subunits with DNA. However, results from Western blotting analysis utilizing nuclear extracts from glioma cells throughout the cell cycle demonstrated that the nuclear accumulation of p50 and p65 perfectly correlates with their DNA-binding activity. These observations suggest that the nuclear translocation of the p50/p65 subunit of NF-kappa B in glioma cells is cell cycle stage-dependent and that is distinct from the differential mRNA expression of these genes during glioma cell cycling. The possible role of NF-kappa B in glioma cell formation and regulation of cellular genes by NF-kappa B in these tumor cells is discussed.