Matriptase-2 inhibits HECV motility and tubule formation in vitro and tumour angiogenesis in vivo

Abstract

The type II transmembrane serine proteases (TTSP) are cell surface proteolytic enzymes that mediate a diverse range of cellular functions, including tumour invasion and metastasis. Matriptase-2 is a member of the TTSP family and has been shown to have a key role in cancer progression. The role of matriptase-2 in angiogenesis and angiogenesis-related cancer progression is currently poorly understood. This study aims to elucidate the role of matriptase-2 in tumour angiogenesis. Matriptase-2 was over-expressed in human vascular endothelial cells, HECV, using a mammalian expression plasmid. The altered cells were used in a number of in vitro and in vivo assays designed to investigate the involvement of matriptase-2 in angiogenesis. Over-expression had no significant effect on the growth and adhesion of HECV cells. However, there was a significant reduction in the motility of the cells and their ability to form tubules in an artificial basement membrane (p < 0.01 for both). HECV(mat2 exp) cells inoculated into CD-1 athymic mice along with either PC-3 prostate cancer cells or MDA-MB-231 breast cancer cells showed a dramatic decrease in tumour development and growth in the prostate tumours (p < 0.01) and a lesser, non-significant, decrease in the breast tumours (p = 0.08). Over-expression of matriptase-2 also decreased urokinase type plasminogen activator total protein levels in HECV and prostate cells. The study concludes that matriptase-2 has the ability to suppress the angiogenic nature of HECV cells in vitro and in vivo. It also suggests that matriptase-2 could have a potential role in prostate and breast tumour suppression through its anti-angiogenic properties.

Liposome encapsulated Disulfiram inhibits NFκB pathway and targets breast cancer stem cellsin vitroandin vivo

“…The breast cancer cell lines MCF7, MDA-MB-231, T47D and normal breast epithelial cell line MCF10A were purchased from ATCC (Middlesex, UK). HeCV, the normal human vascular endothelial cell line, [ 52 ] was kindly provided by Prof W Jiang (Cardiff University, UK). Gemcitabine (dFdC), doxorubicin (Dox), paclitaxel (PTX), copper(II) chloride (CuCl 2 ), copper gluconate (CuGlu), N -acetyl-cysteine (NAC) and poly-2-hydroxyethyl methacrylate (poly-HEMA) were purchased from Sigma (Dorset, UK).…”

Design and chemical syntheses of potent matriptase‐2 inhibitors based on trypsin inhibitor SFTI‐1 isolated from sunflower seeds

“…[2] Members of the TTSPs are in general composed of a single-pass transmembrane domain located near the N-terminus of the protein that separates a short, cytoplasmic domain from a larger extracellular part of the protease containing a stem region and a C-terminal catalytic domain of the chymotrypsin fold. [4][5][6] Additionally, matriptase-2 plays a pivotal role in iron homeostasis. These enzymes possess an intracellular, cytoplasmic domain giving an excellent opportunity to mediate signal transduction between the cell and its extracellular environment and to regulate various cellular responses.…”

Impact of pigment epithelium-derived factor on colorectal cancer in vitro and in vivo

“…Mucinous adenocarcinoma tissue showed more positive tumour expression when compared to adenocarcinoma tissue on immunohistochemical staining, which is consistent with work from Ji et al (2013) where patients with mucinous adenocarcinoma displayed higher PEDF plasma levels than adenocarcinoma patients [ 29 ]. This is perhaps a slightly surprising result, as mucinous adenocarcinoma of the colon or rectum is well recognised to have a poorer survival rate when compared to non-mucinous adenocarcinoma [ 37 ] and hence there must be other influences, yet to be identified, that are specific to mucin-producing tumours that are responsible for the higher expression levels of PEDF.…”

“…Visualisation of products took place on 0.8% agarose gel using SYBR safe DNA gel stain (Invitrogen, Manchester, UK). PEDF transcript levels in the colorectal tissue specimens were determined using real-time qPCR, using Amplifluor™ technology as described before [ 37 ]. In brief, the ampliflour probe consists of a 3′ region specific to the Z-sequence (ACTGAACCTGACCGTACA) present on the target specific primers and a 5′ hairpin structure labelled with a flourophore [ 38 ].…”