Overexpression of urokinase receptor in breast cancer cells results in increased tumor invasion, growth and metastasis

Abstract

We have examined the role of urokinase receptor (uPAR) in tumor invasion and metastasis by developing a homologous model of uPAR overexpression in a rat breast cancer cell line (Mat B III) using gene transfer technique. Control (pRc‐CMV) and experimental plasmid (pRc‐uPAR‐S) were transfected into Mat B III cells by using Lipofectin reagent. Levels of uPAR production were accessed by Northern blotting, immunofluorescence, receptor binding and ELISA. At least 3 experimental clones (pRc‐uPAR‐S), expressing 3‐ to 5‐fold higher levels of uPAR than control (pRc‐CMV), were selected for further analysis. Experimental cells overexpressing uPAR showed a 4‐ to 5‐fold higher invasive capacity compared with control cells in a Boyden chamber invasion assay. Both control and experimental cells (1 × 10⁶ cells) were injected into the mammary fat pad of syngeneic female Fischer rats. Animals were sacrificed at timed intervals and evaluated for the development of tumor growth and metastasis. Animals receiving cells overexpressing uPAR had significantly larger tumor volume and weight throughout our study. Furthermore, due to increased uPAR expression, experimental animals developed large metastatic lesions in liver, spleen and lymph nodes. Our results therefore demonstrate the role of uPAR in tumor progression, due to its ability to localize uPA within the tumor cell milieu. © 1996 Wiley‐Liss, Inc.