Multimodal therapy for synergic inhibition of tumour cell invasion and tumour-induced angiogenesis

Pamela Zengel,Olivier Gires,Brigitte Mack,Alexander Berghaus,Suna Schmitz,Bernd Muehlenweg,Stefan Zahler,Diana Ramp

doi:10.1186/1471-2407-10-92

Abstract

BackgroundSquamous cell carcinoma of the head and neck (SCCHN) are highly invasive tumours with frequent local and distant recurrence. Metastasis formation requires degradation of the extracellular matrix, which is fulfilled by membrane-associated proteases such as the urokinase plasminogen activator (uPA). WX-UK1 is a competitive active site inhibitor of the protease function of uPA that impairs on the capacity of tumour cells to invade in vitro.MethodsIn the present study, effects of combinations of WX-UK1 with matrix metalloprotease inhibitors (MMP, galardin®) and cyclooxygenase-2 (COX-2, celecoxib®) inhibitors on tumour cell proliferation, invasion, and angiogenesis induction were evaluated. Matrigel invasion chambers and a spheroid co-cultivation model with human fibroblast served to determine the invasive potential of both FaDu (SCCHN) and HeLa (cervical carcinoma) cells, each treated with combinations of Celecoxib®, Galardin®, and WX-UK1.ResultsBlocking of single protease systems resulted in a significant 50% reduction of tumour cell invasion using WX-UK1, while the triple combination was even more effective with 80% reduction of invasion. Additionally, a sprouting assay with HUVEC was used to test the anti-angiogenetic potential of the triple combination, resulting in a 40% decrease in the sprouting rate.ConclusionsA combined approach targeting different families of proteases and cyclooxygenases represents a promising adjuvant therapy.

Highlights

Squamous cell carcinoma of the head and neck (SCCHN) are highly invasive tumours with frequent local and distant recurrence
Triple inhibition of urokinase plasminogen activator (uPA), matrix metalloproteases (MMP), and COX-2 decreases the invasive capacity of tumour cells Inhibition of the uPA protease system with WX-UK1 displayed excellent results with a 50% rate of inhibition of tumour cell invasion into matrigel and in a spheroid co-culture model [17]
We chose three mechanisms contributing to tumour cells invasion and neo-angiogenesis to be inhibited in a multimodal fashion, i.e. uPA, matrix metalloproteases, and cyclooxygenase 2

Summary

Introduction

Squamous cell carcinoma of the head and neck (SCCHN) are highly invasive tumours with frequent local and distant recurrence. Metastasis formation requires degradation of the extracellular matrix, which is fulfilled by membrane-associated proteases such as the urokinase plasminogen activator (uPA). Squamous cell carcinoma of the head and neck (SCCHN) are aggressive tumours, which are still associated with poor prognosis despite improvements in surgical and radiotherapeutic techniques [1]. Migration through surrounding tissue is achieved upon the degradation of the extracellular matrix (ECM) by both, membrane-fixed and soluble proteases. In this respect, matrix metalloproteases (MMP) and the urokinase-type plasminogen-activator inhibits protease upon attachment to the active center of the enzyme, resulting in a reduction of tumour expansion [6]. Combinatorial inhibition of MMPs and the plasminogen activator system using siRNA approaches likewise revealed efficient with a 60% and 90% down-regulation of invasion and angiogenesis, respectively [11,12]

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