Effect of cancer procoagulant (CP) on the growth and adhesion of MCF-7 cells to vitronectin in vitro

Abstract 

Cancer procoagulant (CP) is a cysteine protease produced by fetal and malignant tissues, activating in vitro blood coagulation factor X. It has been demonstrated that CP is able to stimulate blood platelet adhesion to fibrinogen and collagen. The pro-adhesive properties of CP could play an important role in metastatic spread of cancer as well as in primary tumor growth.

Effects of anti-CP antibody on the growth of MCF-7 breast cancer cells and on the cells adhesion to vitronectin have been analyzed in vitro. Addition of polyclonal anti-CP antibody to MCF-7 cell culture resulted in 16–18% (P<0.001) decrease in the cells viability as compared with the control (other antibody or no antibody in the culture).

Preincubation of MCF-7 cells with anti-CP antibody reduced the cells adhesion to vitronectin. Further addition of purified CP (0.5–8μg/ml) to the MCF-7 cells preincubated with anti-CP antibody resulted in complete recovery of adhesive properties of the cells. However, when high concentration (16μg/ml) of CP was added to the sample, only partial recovery of the adhesive properties by the cells was observed.

Results of the experiments support the hypothesis that CP is involved in the growth of cancer cells, but its pro-coagulative properties are of secondary importance. One of the possible mechanisms of the interactions between CP and malignant cell could be the regulation of the cell adhesion processes.

Keywords: Cancer procoagulant, Cell adhesion, Vitronectin, MCF-7

Abbreviations: BCA, bicinchoninic acid, BSA, bovine serum albumine, CP, cancer procoagulant, DMEM, Dulbecco's minimum essential medium, DMSO, dimethyl sulphoxide, E-64, trans-epoxysuccinyl-l-leucylamido (4-guanidino)-butane, ECM, extracellular matrix, MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, PAR-1, proteolytically activated receptor-1