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Proteomic analysis of thyrocytes carrying H64 (rs4644) homo-zygotes polymorphism within galectin-3 unveils a carcinogenic profile

Title: Proteomic analysis of thyrocytes carrying H64 (rs4644) homo-zygotes polymorphism within galectin-3 unveils a carcinogenic profile
Authors: Lorenzo Zallocco; Alda Corrado; Maurizio Ronci; Romina Aceto; Maria Rosa Mazzoni; Antonio Lucacchini; Stefano Landi; Federica Gemignani; Laura Giusti
Contributors: Zallocco, Lorenzo; Corrado, Alda; Ronci, Maurizio; Aceto, Romina; Rosa Mazzoni, Maria; Lucacchini, Antonio; Landi, Stefano; Gemignani, Federica; Giusti, Laura
Publisher Information: SIB-Fondazione Alessandro Volta; ITA; Roma
Publication Year: 2021
Collection: CAMPUS Pubblicazioni Scientifiche Unicam (Università di Camerino)
Description: Genomic wide association studies and intensive bioinformatic searches of databases have identified several loci associated with genetic predisposition to thyroid cancer [1,2]. Among these, the LGALS3 gene, which encodes galectin-3 protein (gal-3), is a potential candidate to be considered. Within LGALS3 gene, a common single nucleotide polymorphism (SNP) encoding for the variant Proline (P64) to Histidine (H64) at codon 64 (rs4644) has been found to affect function of galectin-3. The aim of this work was to determine proteomes of rs4644 SNP variants in human thyroid cell line (Nthy-Ori). Nthy-Ori cells have a heterozygote genotype C/A allowing the conversion into homozygote genotypes (either A/A and C/C) using the CRISPR/Cas9 gene editing system, as described by Corrado et al. [3]. Gene edited cells were named ORI-CC and ORI-AA, also called gal-3P64 and gal-3H64, respectively. Proteomic analysis of genetic variants of Nthy-Ori cell was performed using two-dimensional gel electrophoresis coupled to mass spectrometry. A total of about 1300 spots were detected on gels from different samples. Then, after com-putational comparison of images, 37 protein spots were found to be differentially expressed in ORI-AA with comparison to ORI-CC cells. In order to identify the proteins potentially correlated with rs4644 mutation, differentially expressed spots were cut from gels, trypsinized and analyzed by nano-LC ESI MS/MS. Ingenuity pathways analysis (IPA) was performed to recognize molecular and cellular functions correlated to differentially expressed proteins. Some of the identified proteins found differentially expressed in gal-3H64 respect to gal3P64 variants, belong to chaperones class as HSPA6, HSPAB1 and to actin-binding protein class, as the calponin 3. IPA analysis suggested the involvement of cancer-related upstream regulators such as TP53, MYCN, and HIF1A1. For the first time, rs4644 SNP variants in thyroid cell were disclosed by a proteomic analysis approach to cause cancer-related molecular changes in ...
Document Type: conference object
Language: English
Relation: ispartofbook:61° SIB MEETING Virtual Edition; 61° SIB MEETING; firstpage:1; lastpage:483; numberofpages:483; https://hdl.handle.net/11581/454903
Availability: https://hdl.handle.net/11581/454903
Accession Number: edsbas.83AB815F
Database: BASE