Proteomic Analysis ofPTCH1+/− Fibroblast Lysate and Conditioned Culture Media Isolated from the Skin of Healthy Subjects and Nevoid Basal Cell Carcinoma Syndrome Patients

Giovanni Ponti,Cristina Magnoni,Elisa Bellei,Lorenza Pastorino,Giorgia Bertazzoni,Pietro Loschi,Alexander Michael Witkowski,Paola Azzoni,Luisa Benassi,Stefania Bergamini,Annamaria Pollio,Giovanni Pellacani,Aurora Cuoghi,Tiziana Petrachi,Aldo Tomasi,Emanuela Monari

doi:10.1155/2013/794028

Abstract

Background. The pathogenesis underlying the increased predisposition to the development of basal cell carcinomas (BCCs) in the context of Gorlin-Goltz syndrome is linked to molecular mechanisms that differ from sporadic BCCs. Patients with Gorlin syndrome tend to develop multiple BCCs at an early age and present with tumors of non-sun-exposed skin. The aim of this study was to compare the proteomic profile of cultured fibroblast and fibroblast conditioned culture media of PTCH1+ and nonmutated fibroblasts. Results. Proteomic analysis was performed using Surface-Enhanced Laser Desorption/Ionization Time-of-Flight mass spectrometry in PTCH1+ fibroblast conditioned media isolated from not affected sun-protected skin areas of Gorlin patients and from healthy subjects. 12 protein cluster peaks, >5 kDa, had significant differences in their peak intensities between PTCH1+ and PTCH1− subject groups. We detected a strongly MMP1 overexpression in PTCH1+ fibroblasts obtained from NBCCS patients with respect to healthy donors. Conclusion. Protein profiles in the fibroblast conditioned media revealed statistically significant differences between two different types (missense versus nonsense) of PTCH1 mutations. These differences could be useful as signatures to identify PTCH1 gene carriers at high risk for the development of NBCCS-associated malignancies and to develop novel experimental molecular tailored therapies based on these druggable targets.

Highlights

Patients with germ-line mutations in tumor suppressor genes represent an intriguing heredofamilial model of cancer susceptibility and genotype-phenotype correlation [1]
PTCH1 mutations lead to complex syndromes such as the Gorlin syndrome (GS) named nevoid basal cell carcinoma syndrome (NBCCS, OMIM #109400)
We investigated patients with different subtypes of germ-line PTCH1 mutations, through the comparison of both genetic and proteomic profiles

Summary

Introduction

Patients with germ-line mutations in tumor suppressor genes represent an intriguing heredofamilial model of cancer susceptibility and genotype-phenotype correlation [1]. A variety of other benign or malignant tumors can be found in association with these developmental defects, that is, ovarian fibroma, medulloblastoma, rhabdomyosarcomas cardiac fibromas, and ameloblastoma In this hereditary setting, the genotype-phenotype correlation is not always present: gene carriers and family members sharing the same PTCH1 germ line mutation have a variable phenotype [3]. Protein profiles in the fibroblast conditioned media revealed statistically significant differences between two different types (missense versus nonsense) of PTCH1 mutations. These differences could be useful as signatures to identify PTCH1 gene carriers at high risk for the development of NBCCS-associated malignancies and to develop novel experimental molecular tailored therapies based on these druggable targets

Objectives

Methods

Conclusion