title: Inhibition of chemomigration of a human prostatic carcinoma cell (TSU-pr1) line by inhibition of epidermal growth factor receptor function.
abstract: Chemoattractants expressed at bony sites and pelvic lymph nodes are thought to promote the preferential metastasis of human prostate tumor cells to these organs. Epidermal growth factor (EGF) is a potent chemoattractant for several human metastatic prostate tumor cell lines, including the TSU-pr1 cell line, and EGF has been localized to the stroma of both bony sites and pelvic lymph nodes in humans. Hence, we investigated whether the TSU-pr1 cell line expresses a functional EGF receptor (EGFR), which when antagonized reduces EGF-mediated chemomigration of this cell line. In this context, the EGFR immunoprecipitated from cell lysates of TSU-pr1 cells comigrated with the EGFR from A431 cells at a molecular weight of 170 kD. Addition of human EGF (hEGF) to the TSU-pr1 cells for 5 min stimulated the dose-dependent biphasic phosphorylation of the EGFR, with maximal stimulation of EGFR phosphorylation occurring at 2 ng/ml hEGF. In addition, treatment of hEGF-stimulated (2 ng/ml) TSU-pr1 cells with 0.5 microgram/ml anti-hEGF monoclonal antibody or 100 nM staurosporine inhibited EGFR phosphorylation. Conversely, as negative controls, treatment of hEGF-stimulated (2 ng/ml) TSU-pr1 cells with K252a or dimethyl sulfoxide (DMSO) vehicle did not inhibit EGFR phosphorylation. TSU-pr1 cells were stimulated to migration in 4 hr across Boyden chambers in response to 10 ng/ml hEGF. Treatment of the TSU-pr1 cells with anti-hEGFR monoclonal antibody inhibited in a dose-dependent manner the chemomigration of the TSU-pr1 cells across Boyden chambers. Similarly, treatment of the TSU-pr1 cells with staurosporine inhibited in a dose-dependent manner the chemomigration of the TSU-pr1 cells across Boyden chambers. These results demonstrate that antagonists of hEGF-mediated hEGFR phosphorylation also antagonize chemomigration of the TSU-pr1 cells across Boyden chambers, suggesting that antagonists of the EGFR in prostate cancer may be useful in the treatment of metastatic disease.
authors.affiliation: Department of Urology, Tabriz Medical Sciences University, Iran.
authors.forename: A
authors.initials: A
authors.lastname: Zolfaghari
authors.forename: D
authors.initials: D
authors.lastname: Djakiew
mesh_terms: D000470:Alkaloids / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D002227:Carbazoles / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology*; D002277:Carcinoma / Q000473:pathology* / Q000503:physiopathology; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D004815:Epidermal Growth Factor / Q000494:pharmacology / Q000502:physiology; D066246:ErbB Receptors / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000502:physiology; D006801:Humans; D015151:Immunoblotting; D026121:Indole Alkaloids; D008297:Male; D009362:Neoplasm Metastasis; D010766:Phosphorylation; D011471:Prostatic Neoplasms / Q000473:pathology* / Q000503:physiopathology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D019311:Staurosporine; D014407:Tumor Cells, Cultured
chemical_list: D000470:Alkaloids; D000911:Antibodies, Monoclonal; D002227:Carbazoles; D002273:Carcinogens; D004791:Enzyme Inhibitors; D026121:Indole Alkaloids; D004815:Epidermal Growth Factor; C049985:staurosporine aglycone; D066246:ErbB Receptors; D011493:Protein Kinase C; D019311:Staurosporine; D004121:Dimethyl Sulfoxide