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In vitro anti-invasive effects of N-(4-hydroxyphenyl)-retinamide on human prostatic adenocarcinoma
J H Kim1, T Tanabe, G W Chodak
1University of Chicago Hospital, Section of Urology, IL 60637, USA.
Abstract:
Components of malignant invasion, namely cellular adhesion, motility, and proteolytic capability provide potential sites of pharmacological intervention for malignancy. In this study, a series of experiments were performed to examine the effects of N-(4-hydroxyphenyl) retinamide (4-HPR, Fenretinide) on cellular adhesion, motility and proteolytic activity of established prostate cancer cell lines, TSU-PR 1 and PC-3. Radioadhesion study showed that the treatment of TSU-PR 1 and PC-3 cells with 10(-6) M of 4-HPR resulted in a 32% and 37% reduction (p < 0.05), respectively, in the cellular adhesion to the matrigel extract. Radiomigration assay also demonstrated that 4-HPR concentration of 10(-6) M reduced the cellular motility by 29% in TSU-PR1 and 28% in PC-3 cells (p < 0.05). Spectrolyse PL indirect chromogenic assay revealed an increase in total activatable uPA activity (TSU-PR 1: 25%, PC-3: 32%, P < 0.05), while Spectrolyse UK direct assay demonstrated a mild, but a statistically significant reduction (PC-3: 5%, TSU-PR1: 9%, P < 0.05) in active uPA activity. Northern analysis and ELISA assays showed that 4-HPR at 10(-6) M enhances the expression of type 1 plasminogen activator inhibitor (PAI-1). Type IV collagenase western blot analysis and densitometry did not demonstrate suppression of the enzyme secretion, but in fact suggested increased translation of the enzyme when treated with 10(-6) M concentration of fenretinide. The results of this study demonstrate that 4-HPR inhibits in vitro cellular adhesion and motility of human prostate adenocarcinoma cell lines, TSU-PR1 and PC-3. Additionally, uPA and PAI-1 assay results suggest that 4-HPR may impair active uPA's proteolytic activity while upregulating the expression of total activatable uPA and PAI-1. The results of this study therefore support 4-HPR's role as a potential anti-invasive agent.
Insights
N-(4-hydroxyphenyl) retinamide (4-HPR, Fenretinide) significantly reduces prostate cancer cell adhesion and motility. This study suggests 4-HPR is a potential anti-invasive agent by modulating urokinase plasminogen activator and its inhibitor.
Area of Science:
- Oncology
- Pharmacology
- Cell Biology
Background:
- Malignant invasion involves cellular adhesion, motility, and proteolytic activity, offering targets for cancer intervention.
- Prostate cancer cell lines (TSU-PR1 and PC-3) are established models for studying invasion mechanisms.
Purpose of the Study:
- To investigate the effects of N-(4-hydroxyphenyl) retinamide (4-HPR, Fenretinide) on prostate cancer cell adhesion, motility, and proteolytic activity.
- To evaluate 4-HPR's potential as an anti-invasive agent in vitro.
Main Methods:
- Radioadhesion and radiomigration assays were used to assess cellular adhesion and motility.
- Spectrolyse assays, Northern analysis, ELISA, and Western blot were employed to analyze proteolytic activity and related protein expression (uPA, PAI-1, Type IV collagenase).
Main Results:
- 10(-6) M 4-HPR significantly reduced cellular adhesion (32-37%) and motility (28-29%) in TSU-PR1 and PC-3 cells.
- 4-HPR increased total activatable urokinase plasminogen activator (uPA) activity (25-32%) while decreasing active uPA (5-9%).
- 4-HPR enhanced the expression of plasminogen activator inhibitor-1 (PAI-1) and suggested increased translation of Type IV collagenase.
Conclusions:
- 4-HPR effectively inhibits in vitro cellular adhesion and motility of human prostate adenocarcinoma cell lines.
- Results indicate 4-HPR may impair active uPA proteolytic activity and upregulate total activatable uPA and PAI-1 expression.
- 4-HPR demonstrates potential as an anti-invasive agent for prostate cancer treatment.