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Expression of antisense uPAR and antisense uPA from a bicistronic adenoviral construct inhibits glioma cell invasion, tumor growth, and angiogenesis

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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Acknowledgements

We thank Karen Minter for preparation of the manuscript.

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Correspondence to Jasti S Rao.

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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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