The recombinant kringle domain of urokinase plasminogen activator inhibits <i>in vivo</i> malignant glioma growth

Kim, Chung Kwon; Hong, Sung Hee; Joe, Yeonsoo; Shim, Byoung-Shik; Lee, Suk-Keun; Hong, Yong-Kil

doi:10.1111/j.1349-7006.2006.00378.x

Cited by 9 publications

(6 citation statements)

References 18 publications

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“…Our results also show that simultaneous downregulation of uPAR and uPA suppressed angiogenin, a proangiogenic molecule. Recently, researchers have shown that the recombinant kringle domain (UK1) of uPA had antiangiogenic activity by causing the suppression of various proangiogenic molecules, including angiogenin, in glioma cells (35). Our results confirm the previous findings that interference with the uPAR-uPA system retards angiogenesis.…”

Section: Discussionsupporting

confidence: 90%

Suppression of the uPAR–uPA System Retards Angiogenesis, Invasion, and In Vivo Tumor Development in Pancreatic Cancer Cells

Gorantla

Asuthkar

Rao

et al. 2011

Molecular Cancer Research

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Despite existing chemotherapy and surgical resection strategies, pancreatic cancer is one of the major causes of mortality in the United States with a 5-year mean survival rate of less than 5%. The activation of the urokinasetype plasminogen activator receptor-urokinase-type plasminogen activator (uPAR-uPA) system in the development of pancreatic ductal adenocarcinoma has been well established. In the present study, we used 2 pancreatic cancer cell lines, MIA PaCa-2 and PANC-1 to show the effects of uPAR and uPA downregulation. From the results, we observed that RNAi expressing plasmids efficiently downregulated mRNA and protein expression of uPAR and uPA. In vitro and in vivo angiogenic assays revealed a significant decrease in the angiogenic potential of MIA PaCa-2 and PANC-1 cells that were downregulated for both uPAR and uPA. From the angiogenesis antibody array analysis, we observed that the simultaneous downregulation of uPAR and uPA resulted in the downregulation of angiogenin and overexpression of RANTES. Further, FACS analysis showed that the simultaneous downregulation of uPAR and uPA caused the accumulation of cells in the sub-G 0/1 phase in both MIA PaCa-2 and PANC-1 cells. In addition, Western blot analysis revealed that downregulation of uPAR and uPA caused the activation of caspase 8 and CAD, which is indicative of apoptosis, and in vivo TUNEL assay confirmed these results. Finally, we observed the nuclear localization of uPA and that uPA interacts with the transcription factor Lhx-2. Taken together, the results of the present study show that the targeting of the uPARuPA system has therapeutic potential. Mol Cancer Res; 9(4); 377-89. Ó2011 AACR.

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Section: Discussionsupporting

confidence: 90%

Suppression of the uPAR–uPA System Retards Angiogenesis, Invasion, and In Vivo Tumor Development in Pancreatic Cancer Cells

Gorantla

Asuthkar

Rao

et al. 2011

Molecular Cancer Research

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“…6B). Previous studies by Kim CK and collegues [31] have shown that the recombinant kringle domain of ATF inhibits glioma invasion in vivo . To check if uPA ATF, which includes the GFD and the kringle domain, plays a role in angiogenin expression and secretion, we first treated endothelial cells with different concentrations of ruPA for 24 hrs, then treated with ruPA ATF, and assayed for angiogenin expression by western blotting and angiogenin secretion by ELISA.…”

Section: Resultsmentioning

confidence: 94%

“…Kim et al [31] have shown that UK1, the recombinant kringle domain of urokinase plasminogen activator, shows anti-angiogenic activity. In their study, they demonstrated that systemic administration of UK1 (50 mg/kg) suppressed U87 tumor growth in vivo by 80%.…”

Section: Discussionmentioning

confidence: 99%

Suppression of uPA and uPAR Attenuates Angiogenin Mediated Angiogenesis in Endothelial and Glioblastoma Cell Lines

et al. 2010

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BackgroundIn our earlier reports, we showed that downregulation of uPA and uPAR inhibited glioma tumor angiogenesis in SNB19 cells, and intraperitoneal injection of a hairpin shRNA expressing plasmid targeting uPA and uPAR inhibited angiogenesis in nude mice. The exact mechanism by which inhibition of angiogenesis takes place is not clearly understood.Methodology/Principal FindingsIn the present study, we have attempted to investigate the mechanism by which uPA/uPAR downregulation by shRNA inhibits angiogenesis in endothelial and glioblastoma cell lines. uPA/uPAR downregulation by shRNA in U87 MG and U87 SPARC co-cultures with endothelial cells inhibited angiogenesis as assessed by in vitro angiogenesis assay and in vivo dorsal skin-fold chamber model in nude mice. Protein antibody array analysis of co-cultures of U87 and U87 SPARC cells with endothelial cells treated with pU2 (shRNA against uPA and uPAR) showed decreased angiogenin secretion and angiopoietin-1 as well as several other pro-angiogenic molecules. Therefore, we investigated the role of angiogenin and found that nuclear translocation, ribonucleolytic and 45S rRNA synthesis, which are all critical for angiogenic function of angiogenin, were significantly inhibited in endothelial cells transfected with uPA, uPAR and uPA/uPAR when compared with controls. Moreover, uPA and uPAR downregulation significantly inhibited the phosphorylation of Tie-2 receptor and also down regulated FKHR activation in the nucleus of endothelial cells via the GRB2/AKT/BAD pathway. Treatment of endothelial cells with ruPA increased angiogenin secretion and angiogenin expression as determined by ELISA and western blotting in a dose-dependent manner. The amino terminal fragment of uPA down regulated ruPA-induced angiogenin in endothelial cells, thereby suggesting that uPA plays a critical role in positively regulating angiogenin in glioblastoma cells.Conclusions/SignificanceTaken together, our results suggest that uPA/uPAR downregulation suppresses angiogenesis in endothelial cells induced by glioblastoma cell lines partially by downregulation of angiogenin and by inhibition of the angiopoietin-1/AKT/FKHR pathway.

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“…Previously, we have shown that the recombinant kringle domain of urokinase (UK1) is a potent angiogenesis inhibitor and it inhibits tumor growth in a brain tumor model [ 18 , 19 ]. However, potential problems such as poor bioavailability, antigenicity and inconsistency in bioactivity from batch to batch during production of recombinant protein can hamper the application of UK1 to clinical practice [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

Urokinase-derived peptide UP-7 suppresses tumor angiogenesis and metastasis through inhibition of FAK activation

et al. 2018

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The recombinant kringle domain of urokinase (UK1) has been shown to inhibit angiogenesis and brain tumor growth in vivo. To avoid limitations in application due to mass production of recombinant protein, we attempted to develop a novel peptide inhibitor from UK1 sequence consisting of 83 amino acids that contains α-helices, loops and β-sheets. We dissected UK1 sequence to seven peptides based on structure and amino acid characteristics, and examined the anti-angiogenic activities for the constructed peptides. Among the tested peptides, UP-7 most potently inhibited the proliferation and migration of endothelial cells (ECs) in vitro, and also potently inhibited in vivo angiogenesis in the mouse matrigel plug assay. Such anti-angiogenic activities were not exerted by the scrambled peptide. At molecular level, UP-7 inhibited growth factor-induced phosphorylation of FAK and ERK1/2. It also suppressed formation of stress fibers and focal adhesions and also inhibited the attachment and spreading of ECs onto immobilized fibronectin. In a lung cancer animal model xenografted with non-UP-7-sensitive NCI-H460 cells, systemic treatment of UP-7 effectively suppressed tumor growth through inhibition of angiogenesis. Interestingly, breast cancer cells such as LM-MDA-MB-231 cells were moderately sensitive to UP-7 in proliferation differently from other cancer cells. UP-7 also inhibited migration, invasion and FAK phosphorylation of LM-MDA-MB-231 cells. Accordingly, UP-7 potently inhibited lung metastatic growth of LM-MDA-MB-231 cells in an experimental metastasis model. Taken together, these results suggest that novel peptide UP-7 can be effectively used for treatment of breast cancer metastatic growth through inhibition of angiogenesis and invasion.

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The recombinant kringle domain of urokinase plasminogen activator inhibits in vivo malignant glioma growth

Cited by 9 publications

References 18 publications

Suppression of the uPAR–uPA System Retards Angiogenesis, Invasion, and In Vivo Tumor Development in Pancreatic Cancer Cells

Suppression of the uPAR–uPA System Retards Angiogenesis, Invasion, and In Vivo Tumor Development in Pancreatic Cancer Cells

Suppression of uPA and uPAR Attenuates Angiogenin Mediated Angiogenesis in Endothelial and Glioblastoma Cell Lines

Urokinase-derived peptide UP-7 suppresses tumor angiogenesis and metastasis through inhibition of FAK activation

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