In vitro suppression of urokinase plasminogen activator in breast cancer cells - A comparison of two antisense strategies

Arens, Norbert; Gandhari, Mukesh; Bleyl, U.; Hildenbrand, Ralf

doi:10.3892/ijo.26.1.113

Cited by 16 publications

(22 citation statements)

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“…RNAi is a strong tool for silencing the function of specific genes (35,37,38). Several earlier studies have used chemical inhibitors to investigate the role of the uPA system in progress of metastasis (33,35,52,53).…”

Section: Discussionmentioning

confidence: 99%

“…UPA-or uPAR-neutralizing antibodies (33), protease inhibitors (34), urokinase-specific inhibitors (5,35,36), uPA or uPAR antisense oligonucleotides (11,16,37,38), and small molecules that target uPA or uPAR (39 40) are being investigated as possible strategies to block either uPA or uPAR activity, thereby causing the suppression of tumor growth, invasion, and metastasis.…”

mentioning

confidence: 99%

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RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

Pulukuri

Gondi

Lakka

et al. 2005

Journal of Biological Chemistry

245

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The invasive ability of tumor cells plays a key role in prostate cancer metastasis and is a major cause of treatment failure. Urokinase plasminogen activator-(uPA) and its receptor (uPAR)-mediated signaling have been implicated in tumor cell invasion, survival, and metastasis in a variety of cancers. This study was undertaken to investigate the biological roles of uPA and uPAR in prostate cancer cell invasion and survival, and the potential of uPA and uPAR as targets for prostate cancer therapy. uPA and uPAR expression correlates with the metastatic potential of prostate cancer cells. Thus, therapies designed to inhibit uPA and uPAR expression would be beneficial. LNCaP, DU145, and PC3 are prostate cancer cell lines with low, moderate, and high metastatic potential, respectively, as demonstrated by their capacity to invade the extracellular matrix. In this study we utilized small hairpin RNAs (shRNAs), also referred to as small interfering RNAs, to target human uPA and uPAR. These small interfering RNA constructs significantly inhibited uPA and uPAR expression at both the mRNA and protein levels in the highly metastatic prostate cancer cell line PC3. Our data demonstrated that uPA-uPAR knockdown in PC3 cells resulted in a dramatic reduction of tumor cell invasion as indicated by a Matrigel invasion assay. Furthermore, simultaneous silencing of the genes for uPA and uPAR using a single plasmid construct expressing shRNAs for both uPA and uPAR significantly reduced cell viability and ultimately resulted in the induction of apoptotic cell death. RNA interference for uPA and uPAR also abrogated uPA-uPAR signaling to downstream target molecules such as ERK1/2 and Stat 3. In addition, our results demonstrated that intratumoral injection with the plasmid construct expressing shRNAs for uPA and uPAR almost completely inhibited established tumor growth and survival in an orthotopic mouse prostate cancer model. These findings uncovered evidence of a complex signaling network operating downstream of uPA-uPAR that actively advances tumor cell invasion, proliferation, and survival of prostate cancer cells. Thus, RNA interference-directed targeting of uPA and uPAR is a convenient and novel tool for studying the biological role of the uPA-uPAR system and raises the potential of its application for prostate cancer therapy.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

Pulukuri

Gondi

Lakka

et al. 2005

Journal of Biological Chemistry

245

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“…To determine the proliferation rate in breast cancer cells, we have established a uPA-suppressed model by two approaches, they include transfection of an antisense-vector construct and siRNA transfection of MDA-MB-231 breast cancer cells (17). Our results indicate that suppression of uPA (65%) by siRNA transfection is associated with a strong decrease of proliferation rate with maximum at the 4th and the 5th day after transfection (38%) (Fig.…”

Section: Discussionmentioning

confidence: 88%

“…Stable transfection of MDA-MB-231 was performed with a pCMVdriven uPA-antisense vector construct (17) and an empty pCMV vector (negative control) using a transfection reagent (FuGENE 6, Roche, Mannheim, Germany). The transfection was performed with a vector:FuGENE ratio of 1:3 (w/v).…”

Section: Methodsmentioning

confidence: 99%

Urokinase-type plasminogen activator induces proliferation in breast cancer cells

Gandhari

Arens²,

Majety³

et al. 2006

Int J Oncol

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Abstract. Urokinase-type plasminogen activator (uPA) is implicated in various pathophysiological processes, including extracellular matrix turnover, cell migration and invasion. Our study aimed to determine the role of uPA in both proliferation and mitogen-activated protein kinase (MAPK) pathway. Hence, we analyzed the effects induced by exogeneous addition of domain-specific uPA antibodies and uPAinteracting molecules on proliferation of uPA-suppressed MDA-MB-231 breast cancer cells. uPA expression was reduced to 53% by stable transfection with an antisense/ vector construct and to 65% by siRNA transfection. Immunocytochemical Ki67 staining and flow cytometry (S-phase) analysis indicated a strong decrease of cellular proliferation activity (35% and 38%, respectively). Exogenous addition of high molecular weight-uPA (HMW-uPA) or incubation with the amino terminal fragment (ATF), which lacks the enzymatic activity of uPA, lead to increased cell proliferation. A strong increase of proliferation was absent when the monoclonal antiuPAR antibody IIIF10 (blocking uPA binding site), soluble uPAR (scavenger effect) and phosphatidyl-inositol-specific phopholipase C (PI-PLC, degrading uPAR) was added prior to the addition of HMW-uPA. In conclusion, HMW-uPA and ATF induce proliferation of breast cancer cells by binding to uPAR. Thereby, integrins situated adjacent to uPAR carry the signals into the cell, thus stimulating proliferation that is mediated via the MAPK pathway.

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“…Recently emerging as an important tool for silencing specific genes, RNAi is more potent in inhibiting gene expression than other methods and carries less risk of toxicity [21][22][23] . More importantly, with the effectiveness and relative ease with which dsRNA can be introduced into cells and tissues, RNAi has a significant therapeutic potential for targeting human carcinomas.…”

Section: Discussionmentioning

confidence: 99%

The role of Med19 in the proliferation and tumorigenesis of human hepatocellular carcinoma cells

Zou

Wang

et al. 2011

Acta Pharmacol Sin

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In addition, the ability of the Med19-RNAi-Lentivirus vector-infected Hep3B cells to form tumors after inoculation into nude mice was determined. Results: Recombinant lentiviral vectors expressing small interfering RNA (siRNA) against Med19 were constructed and were found to efficiently downregulate Med19 mRNA and protein levels in HepG2 and Hep3B cells. Furthermore, the inhibition of Med19 by RNAi dramatically reduced hepatocellular carcinoma cell proliferation, induced cell-cycle arrest in the G 0 /G 1 phase, and suppressed tumor formation. Conclusion: These results provide new evidence of an important role for Med19 in the development of hepatocellular carcinomas, suggesting that lentivirus-mediated RNAi to target Med19 is a potential tool for inhibiting cancer cell proliferation and tumorigenesis.

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In vitro suppression of urokinase plasminogen activator in breast cancer cells - A comparison of two antisense strategies

Cited by 16 publications

References 0 publications

RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

RNA Interference-directed Knockdown of Urokinase Plasminogen Activator and Urokinase Plasminogen Activator Receptor Inhibits Prostate Cancer Cell Invasion, Survival, and Tumorigenicity in Vivo

Urokinase-type plasminogen activator induces proliferation in breast cancer cells

The role of Med19 in the proliferation and tumorigenesis of human hepatocellular carcinoma cells

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