Finasteride interfere with MMP2 and MMP9 expression profile on human prostate cancer PC-3 cell line


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Andrei Moroz1, Flavia Delella1, Sergio Santos1, Jaqueline Rinaldi1, Andrelson Rinaldi4, Elenice Deffune2, Hernandes Carvalho3, Sergio Felisbino1
1Biosciences Institute, Department of Morphology - Sao Paulo State University (UNESP), Botucatu, SP, Brazil,2Blood Transfusion Center, Cell Engineering Laboratory - Sao Paulo State University (UNESP), Botucatu, SP, Brazil,3Biology Institute, Department of Cell Biology, Campinas State University (UNICAMP), Campinas, SP, Brazil,4Grande Dourado Federal University, Exact Sciences and Technology School (UFGD), Grande Dourado, MS, Brazil

Background

Finasteride (FIN), a 5 alpha-reductase inhibitor, was recently approved as chemoprevention therapy for higher risk prostate cancer individuals. However, little is know about FIN effects on androgen-independent prostate cancers that are more aggressive and metastasizing, mainly using higher activity of matrix metalloproteinases (MMPs). We evaluated the FIN effects on MMP2 and MMP9 expression of PC-3, an androgen-independent epithelial prostate cancer cell line and to determine the best FBS concentration to perform MMP analysis.

Method

PC-3 cells were plated at 3x104cels/cm2 and expanded until 80% confluence. Cells were then exposed in duplicate to 50µM FIN or DMSO 0.1% for 24 hours in different FBS concentrations (10%, 1% and 0), viability assessed and collected. Afterwards, cells at 80% of confluence were cultivated on 1% FBS plus 50µM FIN or DMSO 0.1% for 24 hours, fixed and submitted to immunohistochemistry for MMP2 and MMP9.

Results

At 10% FBS concentration FIN induced 7.39% viability decrease. On lower hormonal conditions FIN showed higher cell-toxicity: 11.59% viability decrease at 1% FBS and 9.72% at FBS free medium. Regarding MMPs expression, only 24 hours of FIN exposure were sufficient to invert the MMP2/MMP9 expression profile. Before exposure cells expressed MMP2 and no MMP9. After exposure cells ceased MMP2 expression and started expressing MMP9.

Conclusion

Previously, we observed, by gelatin zymography, that both FBS has MMP-2 and MMP-9 activity. So, a minimal and optimal FBS concentration should be determined to access FIN effects on PC-3 MMPs expression, to eliminate FBS-derived contaminants in MMPs activity assay. Our results suggest that 1% FBS provides both lower exogenous MMP contaminants and allow studying prolonged FIN effects on MMP activity and cell viability. FIN showed potential to interfere with MMPs expression profile, as seen with a shift in MMP2 to MMP9 production, confirming previous experiments in Wistar rats.