Downregulation of the urokinase-type plasminogen activator receptor through inhibition of translation by antisense oligonucleotide suppresses invasion of human glioblastoma cells.
We previously showed that downregulation of the urokinase-type plasminogen activator receptor (uPAR) in the SNB19 human glioblastoma cell line by the stable transfection of a plasmid expressing a 300 bp antisense sequence to the 5' end of the uPAR gene produced a decrease in the amount of target mRNA. In a more recent study, we found that adenovirus-mediated transduction (Ad-uPAR) of the same uPAR antisense gene construct in SNB19 cells also downregulated uPAR protein levels. We report here that Ad-uPAR-transfected SNB19 cells produced the same amounts of target uPAR mRNA but significantly less protein by in vitro translation and by in situ [35S] labeling compared to Ad-CMV vector-transfected and mock-transfected cells. This antisense construct also inhibited glioblastoma cell invasion confirming previous results. We conclude that downregulation of uPAR by this antisense gene construct results from inhibition of protein translation.
Mohan
P M
PM
Department of Neurosurgery, The University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
Lakka
S S
SS
Mohanam
S
S
Kin
Y
Y
Sawaya
R
R
Kyritsis
A P
AP
Nicolson
G L
GL
Rao
J S
JS
D003094:Collagen; D016373:DNA, Antisense / Q000494:pharmacology*; D015536:Down-Regulation / Q000187:drug effects*; D004338:Drug Combinations; D005909:Glioblastoma / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D007797:Laminin; D009361:Neoplasm Invasiveness / Q000503:physiopathology / Q000517:prevention & control*; D009363:Neoplasm Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D014176:Protein Biosynthesis / Q000187:drug effects*; D011509:Proteoglycans; D016372:RNA, Antisense / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000378:metabolism; D012334:RNA, Neoplasm / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000096:biosynthesis* / Q000235:genetics / Q000502:physiology; D055293:Receptors, Urokinase Plasminogen Activator; D014162:Transfection; D014407:Tumor Cells, Cultured
D016373:DNA, Antisense; D004338:Drug Combinations; D007797:Laminin; D009363:Neoplasm Proteins; C524898:PLAUR protein, human; D011509:Proteoglycans; D016372:RNA, Antisense; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; C059485:matrigel; D003094:Collagen