Effect of lysine antifibrinolytics and cyclooxygenase inhibitors on the proteolytic profile of breast cancer cells interacting with macrophages or endothelial cells

Abstract

Extracellular matrix (ECM) proteases play a key role in the regulation of tumour invasion, growth, and transendothelial migration. The expression of ECM proteases and their endogenous inhibitors by cancer cells is regulated by stromal cells. We investigated the effect of commonly used perioperative medications on this regulation. Breast cancer cells (4T1) were cultured alone or with endothelial cells (H5V) or macrophages (RAW264.7). Cell grown alone or in cocultures were treated with clinically relevant concentrations of cyclooxygenase (COX) inhibitors, aspirin (ASA), ketorolac, celecoxib, or lysine antifibrinolytics, ɛ-aminocaproic acid (EACA) and tranexamic acid (TXA). We determined the level of the ECM proteases urokinase-like plasminogen activator (uPA), matrix metalloproteinase (MMP)-2 and MMP-9, and endogenous MMP inhibitors, tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 in the conditioned media. Antifibrinolytics and COX inhibitors exerted a complex effect on cells grown alone and in cocultures. EACA increased the activity of MMP-9 and TIMP-1 in cocultures of 4T1 and RAW264.7. TXA increased TIMP-1 in the coculture without affecting MMP-9. EACA and TXA both attenuated MMP-2 detected in 4T1 and H5V cocultures. ASA and ketorolac both decreased the activity of MMP-2, MMP-9, and uPA. Celecoxib increased the activity of TIMP-1 in cocultures of 4T1 with both macrophages and endothelial cells. Antifibrinolytics and COX inhibitors can affect the proteolytic profile of the tumour microenvironment. Animal and clinical investigations are warranted to assess the effect of these proteolytic changes on the outcome of cancer surgery.