Inhibition of chemomigration of a human prostatic carcinoma cell (TSU-pr1) line by inhibition of epidermal growth factor receptor function.
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.
Zolfaghari
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Department of Urology, Tabriz Medical Sciences University, Iran.
Djakiew
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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
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