Nafamostat mesilate negatively regulates the metastasis of triple-negative breast cancer cells

Abstract

Triple-negative breast cancer (TNBC) lacking of oestrogen receptor, progesterone receptor, and epidermal growth factor receptor type 2 is a highly malignant disease which results in a poor prognosis and rare treatment options. Despite the use of conventional chemotherapy for TNBC tumours, resistance and short duration responses limit the treatment efficacy. Therefore, a need exists to develop a new chemotherapy for TNBC. The aim of this study was to examine the anti-cancer effects of nafamostat mesilate (NM), a previously known serine protease inhibitor and highly safe drug on breast cancer cells. Here, we showed that NM significantly inhibits proliferation, migration, and invasion in MDA-MB231 cells, induces G2/M phase cell-cycle arrest, and inhibits the expression of cyclin-dependent kinase 1 (CDK1). Exposure of MDA-MB231 cells to NM also resulted in decreased transcription factor activities accompanied by the regulated phosphorylation of signalling molecules and a decrease in metalloproteinases, the principal modulators of the extracellular environment during cancer progression. Especially, inhibition of TGFβ-stimulated Smad2 phosphorylation and subsequent metastasis-related gene expression, and downregulation of ERK activity may be pivotal mechanisms underlying inhibitory effects of NM on NM inhibits lung metastasis of breast cancer cells and growth of colonized tumours in mice. Taken together, our data revealed that NM inhibits cell growth and metastasis of TNBC cells and indicated that NM is a multi-targeted drug that could be an adjunct therapy for TNBC treatment.

Imidazole Antifungal Drugs Inhibit the Cell Proliferation and Invasion of Human Breast Cancer Cells

“…To remove cellular debris, the cells were washed with 1 mL of phosphate buffered saline (PBS), placed in 2 mL of medium and then treated with 100 μM of CTZ, KCZ, FCZ, or ICZ. After 8-h incubation, the widths of the scratched line were measured and compared to that at 0 h ( Mander et al ., 2018 ).…”

“…The inserts were then air-dried and dissolved in 200 μL methanol, and 100 μL dye/solute mixture was transferred to a 96-well plate. The invasion ability of the cells was detected by measuring the absorbance at 540 nm using a Biotek ELISA reader (Fisher Scientific) ( Mander et al ., 2018 ).…”

“…The gel was washed twice alternately with renaturing buffer (2.5% Triton X-100, v/v) and distilled water and then incubated with developing buffer (50 mM Tris-Base, 150 mM NaCl, 1 μM ZnCl 2 , 0.02% NaN 3 , 5 mM CaCl 2 •2H 2 O, pH 7.5) at 37°C for 48 h. The gel was stained with 0.5% Coomassie blue for 1 h and destained with destaining buffer (5% acetic acid in 10% methanol, v/v). The band density was measured using ImageJ 1.45s software (National Institutes of Health [NIH], Bethesda, MD, USA) ( Mander et al ., 2018 ).…”

Regulation of post-translational modification in breast cancer treatment

“…The molecular subtype of breast cancer based on receptor expression is helpful for prognosis and therapeutic response. Four common molecular subtypes are associated with breast cancers, Luminal A (ERα+/PgR+, HER2−), Luminal B (ERα+/PgR+, HER2+), HER2 (ERα−/PgR−, HER2+), and triple-negative (TN, ERα−/PgR−, HER2−) (2, 3). The luminal breast cancer subtypes are characterized by the expression of a set of ERα-related genes and represent approximately 75% of breast cancer patients in postmenopausal women (1).…”

The Molecular Aspect of Antitumor Effects of Protease Inhibitor Nafamostat Mesylate and Its Role in Potential Clinical Applications

“…Subsequently, NM has been proven to downregulate the expression of both matrix metalloproteinase−2 (MMP-2) and−9, vascular endothelial growth factor (VEGF) and transforming growth factor beta 1 (TGF-β1), secreted by tumor cells in head and neck squamous cell carcinoma, and NM inhibits angiogenesis and tumor invasion (9). Furthermore, Mander et al reported the NM significantly inhibits proliferation, migration, and invasion in triple-negative breast cancer, which was identified both in vitro and in vivo (10). Moreover, NM reverses immune resistance induced by interferon-gamma (IFN-ɤ) as a method of increasing programmed cell death ligand-1 (PD-L1) expression in lung and pancreatic cancer (11).…”