Stable transfection of urokinase-type plasminogen activator antisense construct modulates invasion of human glioblastoma cells
Φόρτωση...
Ημερομηνία
Συγγραφείς
Mohanam, S.
Jasti, S. L.
Kondraganti, S. R.
Chandrasekar, N.
Kin, Y.
Fuller, G. N.
Lakka, S. S.
Kyritsis, A. P.
Dinh, D. H.
Olivero, W. C.
Τίτλος Εφημερίδας
Περιοδικό ISSN
Τίτλος τόμου
Εκδότης
Περίληψη
Τύπος
Είδος δημοσίευσης σε συνέδριο
Είδος περιοδικού
peer-reviewed
Είδος εκπαιδευτικού υλικού
Όνομα συνεδρίου
Όνομα περιοδικού
Clin Cancer Res
Όνομα βιβλίου
Σειρά βιβλίου
Έκδοση βιβλίου
Συμπληρωματικός/δευτερεύων τίτλος
Περιγραφή
The diffuse and extensive infiltration of malignant gliomas into the surrounding normal brain is believed to rely on modifications of the proteolysis of extracellular matrix components. A key molecule in regulating plasminogen-mediated extracellular proteolysis is the urokinase-type plasminogen activator (uPA). To investigate the role of uPA in the invasive process of brain tumors, we stably transfected a human glioblastoma cell line SNB19 with a vector capable of expressing an antisense transcript complementary to the 1020 bases at the 3' end of the uPA cDNA. Parental, vector-, and antisense construct-stably transfected cell lines were analyzed for uPA mRNA transcript by Northern blot analysis, for uPA enzyme activity by zymography, and for uPA protein levels by Western blotting. The levels of uPA mRNA, protein, and enzyme activities were significantly lower in antisense clones than in parental and vector controls. Radioreceptor binding studies demonstrated that uPA receptor levels remained the same in parental, vector-, and antisense-transfected cells. The antisense-transfected cells showed a markedly lower level of invasion in the Matrigel invasion assays, and their spheroids failed to invade the fetal rat brain aggregates in the coculture system. Green fluorescent protein (GFP) expressing parental and antisense transfectants was generated for detection in mouse brain tissue without any posttreatment. Intracerebral injection of antisense stable transfectants significantly reduced tumor formation compared with that in controls. Our results suggested that down-regulation of uPA expression may be a feasible approach to reducing the malignancy and invasiveness of glial tumors.
Περιγραφή
Λέξεις-κλειδιά
Animals, Blotting, Northern, Blotting, Western, Brain/embryology/pathology, DNA, Antisense/*genetics, Fibrin/metabolism, Gene Therapy/methods, Glioblastoma/genetics/pathology/*therapy, Humans, Mice, Mice, Nude, Microscopy, Confocal, Neoplasm Invasiveness/genetics/prevention & control, Protein Binding, RNA, Messenger/genetics/metabolism, Rats, Rats, Sprague-Dawley, Transfection, Tumor Cells, Cultured, Urokinase-Type Plasminogen Activator/*genetics/metabolism, Xenograft Model Antitumor Assays
Θεματική κατηγορία
Παραπομπή
Σύνδεσμος
http://www.ncbi.nlm.nih.gov/pubmed/11489835
http://clincancerres.aacrjournals.org/content/7/8/2519.full.pdf
http://clincancerres.aacrjournals.org/content/7/8/2519.full.pdf
Γλώσσα
en
Εκδίδον τμήμα/τομέας
Όνομα επιβλέποντος
Εξεταστική επιτροπή
Γενική Περιγραφή / Σχόλια
Ίδρυμα και Σχολή/Τμήμα του υποβάλλοντος
Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής