Abstract
Urokinase-type plasminogen activator (uPA) and its receptor (uPAR) play an important role in the invasiveness of gliomas and other infiltrative tumors. In glioma cell lines and tumors, high grade correlates with increased expression of uPAR and uPA. We report here the downregulation of uPAR and uPA by delivery of antisense sequences of uPAR and uPA in a single adenoviral vector, Ad-uPAR-uPA (Ad, adenovirus). The bicistronic construct (Ad-uPAR-uPA) infected glioblastoma cell line had significantly reduced levels of uPAR, uPA enzymatic activity and immunoreactivity for these proteins when compared to controls. The Ad-uPAR-uPA infected 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. Intracranial injection of SNB19 cells with the Ad-uPAR-uPA antisense bicistronic construct showed inhibited invasiveness and tumorigenicity. Subcutaneous injections of bicistronic antisense constructs into established tumors (U87 MG) caused regression of those tumors. Our results support the therapeutic potential of targeting the individual components of the uPAR-uPA system by using a single adenovirus construct for the treatment of glioma and other invasive cancers.
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Abbreviations
- uPA:
-
urokinase-type plasminogen activator
- Urokinase-type plasminogen receptor (uPAR); Ad:
-
adenovirus
- CMV:
-
cytomegalovirus
- BGH:
-
bovine growth hormone
- SV40:
-
simian virus type 40
- PCR:
-
polymerase chain reaction
- MOI:
-
multiplicities of infection
- PFU:
-
plaque-forming units
- PBS:
-
phosphate-buffered saline
- FITC:
-
fluorescein-5-isothiocyanate
- DiI:
-
1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate
- DiO:
-
3,3′-dioctadecyloxacarbocyanine perchlorate
- GFP:
-
green fluorescent protein
- ECM:
-
extracellular matrix
- PAR:
-
plasminogen activator receptor
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We thank Karen Minter for preparation of the manuscript.
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Supported by National Cancer Institute Grant CA 75557 (to JSR)
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Gondi, C., Lakka, S., Yanamandra, N. et al. Expression of antisense uPAR and antisense uPA from a bicistronic adenoviral construct inhibits glioma cell invasion, tumor growth, and angiogenesis. Oncogene 22, 5967–5975 (2003). https://doi.org/10.1038/sj.onc.1206535
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DOI: https://doi.org/10.1038/sj.onc.1206535
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