Adenovirus-mediated expression of antisense urokinase plasminogen activator receptor and antisense cathepsin B inhibits tumor growth, invasion, and angiogenesis in gliomas.
We have shown previously that urokinase plasminogen activator receptor (uPAR) and cathepsin B are overexpressed during glioma progression, particularly at the leading edge of the tumor. In the present study, we simultaneously down-regulated uPAR and cathepsin B in SNB19 glioma cell monolayer or SNB19 spheroids using an adenoviral vector carrying antisense uPAR and antisense cathepsin B and a combination of these genes as determined by Western blot analysis. The Ad-uPAR-Cath B-infected cells revealed a marked reduction in tumor growth and invasiveness as compared with the parental and vector controls. In vitro and in vivo angiogenic assays demonstrated inhibition of capillary-like structure formation and microvessel formation after Ad-uPAR-Cath B infection of SNB19 cells when compared with Ad-cytomegalovirus (CMV)-infected or mock-infected controls. Furthermore, using a near infrared fluorescence probe, in vivo imaging for cathepsin B indicated low/undetectable levels of fluorescence after injection of the Ad-uPAR-Cath B construct into pre-established s.c. tumors as compared with Ad-CMV-treated and untreated tumors. The effect with bicistronic construct (Ad-uPAR-Cath B) was much higher than with single (Ad-uPAR/Ad-Cath B) constructs. These results indicate that the down-regulation of cathepsin B and uPAR plays a significant role in inhibiting tumor growth, invasion, and angiogenesis. Hence, the targeting of these two proteases may be a potential therapy for brain tumors and other cancers.
Gondi
Christopher S
CS
Program of Cancer Biology and Department of Biomedical and Therapeutic Sciences, University of Illinois College of Medicine, Peoria, 61656, USA.
Lakka
Sajani S
SS
Yanamandra
Niranjan
N
Olivero
William C
WC
Dinh
Dzung H
DH
Gujrati
Meena
M
Tung
C H
CH
Weissleder
Ralph
R
Rao
Jasti S
JS
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002401:Cathepsin B / Q000037:antagonists & inhibitors* / Q000235:genetics; D002455:Cell Division / Q000235:genetics; D002465:Cell Movement / Q000235:genetics; D016373:DNA, Antisense / Q000008:administration & dosage / Q000235:genetics*; D005260:Female; D015316:Genetic Therapy / Q000379:methods; D005909:Glioblastoma / Q000098:blood supply / Q000235:genetics* / Q000628:therapy* / Q000821:virology; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009361:Neoplasm Invasiveness; D009389:Neovascularization, Pathologic / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors* / Q000235:genetics; D055293:Receptors, Urokinase Plasminogen Activator; D018874:Spheroids, Cellular; D023041:Xenograft Model Antitumor Assays
D016373:DNA, Antisense; C524898:PLAUR protein, human; C524899:Plaur protein, mouse; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D002401:Cathepsin B