Down-regulation of the PTTG1 proto-oncogene contributes to the melanoma suppressive effects of the cyclin-dependent kinase inhibitor PHA-848125.

Abstract

We previously demonstrated that PHA-848125, a cyclin-dependent kinase inhibitor presently under Phase II clinical investigation, impairs melanoma cell growth. In this study, gene expression profiling showed that PHA-848125 significantly modulated the expression of 128 genes, predominantly involved in cell cycle control, in the highly drug-sensitive GL-Mel (p53 wild-type) melanoma cells. Up-regulation of 4 selected genes (PDCD4, SESN2, DDIT4, DEPDC6), and down-regulation of 6 selected genes (PTTG1, CDC25A, AURKA, AURKB, PLK1, BIRC5) was confirmed at protein levels. The same protein analysis performed in PHA-848125-treated M10 melanoma cells - p53 mutated and less sensitive to the drug than GL-Mel cells - revealed no DEPDC6 expression and no changes of PTTG1, PDCD4 and BIRC5 levels. Upon PHA-848125 treatment, a marked PTTG1 down-modulation was also observed in A375 cells (p53 wild-type) but not in CN-Mel cells (p53 mutated). PTTG1 silencing significantly inhibited melanoma cell proliferation and induced senescence, with effects less pronounced in p53 mutated cells. PTTG1 silencing increased PHA-848125 sensitivity of p53 mutated cells but not that of A375 or GL-Mel cells. Accordingly, in M10 but not in A375 cells a higher level of senescence was detected in PHA-848125-treated/PTTG1-silenced cells with respect to PHA-848125-treated controls. In A375 and GL-Mel cells, TP53 silencing attenuated PHA-848125-induced down-modulation of PTTG1 and decreased cell sensitivity to the drug. These findings indicate that PHA-848125-induced down-regulation of PTTG1 depends, at least in part, on p53 function and contributes to the antiproliferative activity of the drug. Our study provides further molecular insight into the antitumor mechanism of PHA-848125.

• Authors: Simona Caporali, Ester Alvino, Lauretta Levati, Alessia I Esposito, Marina Ciomei, Maria G Brasca, Donatella Del Bufalo, Marianna Desideri, Enzo Bonmassar, Ulrich Pfeffer, Stefania D'Atri
• Journal: Biochemical pharmacology (Biochem Pharmacol) Vol. 84 Issue 5 Pg. 598-611 (Sep 01 2012) ISSN: 1873-2968 [Electronic] England
• PMID: 22704958 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2012 Elsevier Inc. All rights reserved.

Chemical References

• MAS1 protein, human
• N,1,4,4-tetramethyl-8-((4-(4-methylpiperazin-1-yl)phenyl)amino)-4,5-dihydro-1H-pyrazolo(4,3-h)quinazoline-3-carboxamide
• Neoplasm Proteins
• Protein Kinase Inhibitors
• Proto-Oncogene Mas
• Pyrazoles
• Quinazolines
• RNA, Small Interfering
• Securin
• pituitary tumor-transforming protein 1, human
• Cyclin-Dependent Kinases

Topics

• Cell Division (drug effects)
• Cell Line, Tumor
• Cyclin-Dependent Kinases (antagonists & inhibitors)
• Down-Regulation (drug effects)
• Gene Regulatory Networks
• Humans
• Melanoma (genetics, pathology)
• Neoplasm Proteins (genetics)
• Protein Kinase Inhibitors (pharmacology)
• Proto-Oncogene Mas
• Proto-Oncogenes
• Pyrazoles (pharmacology)
• Quinazolines (pharmacology)
• RNA, Small Interfering
• Securin