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J Biol Chem, Vol. 274, Issue 22, 15407-15414, May 28, 1999
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From the Aberrant expression of the potent angiogenic
cytokine, vascular endothelial growth factor (VEGF), has been
demonstrated to be associated with most human solid tumors. Both
transcriptional and post-transcriptional mechanisms have been shown to
modulate VEGF expression in a multitude of cell types. Here we report
that when protein kinase C (PKC) pathways were activated in human
glioblastoma U373 cells by phorbol 12-myristate 13-acetate (PMA), VEGF
mRNA expression was up-regulated via a post-transcriptional
mRNA stabilization mechanism. PMA treatment exhibited no increase
in VEGF-specific transcriptional activation as determined by run-off
transcription assays and VEGF promoter-luciferase reporter assays.
However, PMA increased VEGF mRNA half-life from 0.8 to 3.6 h
which was blocked by PKC inhibitors but not by protein kinase A or
cyclic nucleotide-dependent protein kinase inhibitors. When
U373 cells were transfected with antisense oligonucleotide sequences to
the translation start sites of PKC- Departments of Pathology, Beth Israel
Deaconess Medical Center and Harvard Medical School,
Boston, Massachusetts 02215 and the § Department of
Physiology, University of Connecticut Health Center,
Farmington, Connecticut 06030
, -
, -
, -
, -
, or -
isoforms, both PKC- and - antisense oligonucleotides showed
substantial inhibition of PMA-induced VEGF mRNA. In addition,
overexpression of PKC- resulted in a strong constitutive
up-regulation of VEGF mRNA expression. This study demonstrates for
the first time that specific PKC isoforms regulate VEGF mRNA
expression through post-transcriptional mechanisms.
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