FXYD5/Dysadherin, a Biomarker of Endometrial Cancer Myometrial Invasion and Aggressiveness: Its Relationship With TGF-β1 and NF-κB Pathways.

María José Besso,Roxana Schillaci,Lara Lapyckyj,Eva Colas,Cristian Pablo Moiola,Mónica Hebe Vazquez-Levin,María Florencia Mercogliano,Jaume Reventos,Antonio Gil-Moreno,Alejandra Wernicke,Marina Rosso,Roberto Orti,María Laura Matos

doi:10.3389/fonc.2019.01306

Abstract

Objective: Endometrial cancer (EC) is the second most common gynecological cancer worldwide. Myometrial invasion (MI) is a key event in EC dissemination. This study aimed to evaluate FXYD5/dysadherin (FXYD5/Dys) expression in EC tissue and uterine aspirate (UA) biopsies and to assess molecular/functional changes associated with its expression in cellular models.Methods: FXYD5/Dys messenger RNA (mRNA) levels were determined in EC tissue and UA biopsies. FXYD5/Dys expression was evaluated in EC RNAseq data from The Cancer Genome Atlas (TCGA) and GENEVESTIGATOR tools. FXYD5/Dys impact on E-cadherin expression and cell behavior was assessed in EC Hec1a cells treated with transforming growth factor (TGF)-β1, stably transfected with ETV5, and transiently transfected with FXYD5/Dys small interfering RNA (siRNA) or pcDNA3-FXYD5/Dys plasmid.Results: FXYD5/Dys was associated with EC aggressiveness, finding high mRNA levels in tumors depicting MI > 50%, Grade 3, and intermediate/high risk of recurrence. FXYD5/Dys was highly expressed at the tumor invasive front compared to the superficial area. Most results were recapitulated in UA biopsies. FXYD5/Dys modulation in Hec1a cells altered cell migration/adhesion and E-cadherin expression. TGF-β1 treatment of Hec1a cells induced FXYD5/Dys expression. TCGA-UCEC RNAseq analysis revealed a positive correlation between FXYD5/Dys, TGF-β1, and plasminogen activator inhibitor (PAI)-1 mRNA levels. FXYD5/Dys induced nuclear factor (NF)-κB pathway activation in Hec1a cells. FXYD5/Dys mRNA levels positively correlated with transcriptional activation of NF-κB p65-regulated genes. Survival analysis revealed patient segregation into low- and high-risk groups, the latter depicting the highest FXYD5/Dys, PAI-1, tumor necrosis factor (TNF)-α, and TGF-β1 mRNA levels and shorter survival rates.Conclusion: FXYD5/Dys is a novel biomarker of EC progression related to TGF-β1 and NF-κB pathways that collectively promote tumor dissemination and result in poor patient prognosis.

Highlights

Endometrial cancer (EC) is the second most common gynecological neoplasm and the fourth most frequent women cancer worldwide
The Cancer Genome Atlas (TCGA)-UCEC RNAseq analysis revealed a positive correlation between FXYD5/Dys, transforming growth factor (TGF)-β1, and plasminogen activator inhibitor (PAI)-1 messenger RNA (mRNA) levels
FXYD5/Dys mRNA levels positively correlated with transcriptional activation of nuclear factor (NF)-κB p65-regulated genes

Summary

Introduction

Endometrial cancer (EC) is the second most common gynecological neoplasm and the fourth most frequent women cancer worldwide. While Stage IA tumors (International Federation of Gynecology and Obstetrics, FIGO, 2009 classification) display 50% MI (deep MI), the latter associated with poor prognosis (5-years survival rate: Stage IA, 90%, Stages IB–IV, 78–21%) [4]. Routine EC diagnosis involves preoperative anatomopathological endometrial biopsy evaluation coupled to imaging techniques, adjusted at evaluation of the surgical piece. Among 75% tumors preoperatively classified as early stage EC (FIGO Stage I), ∼20% are surgically reclassified as advanced-stage EC (FIGO Stages II–IV). 30% of EC cases are diagnosed when the tumor has invaded >50% of the myometrial wall [5]. This classification is critical for therapeutic management and greatly impacts post-surgery patient morbidity. There are no established molecular biomarkers to determine deep MI and/or to assist in risk stratification in a sensitive, objective, and reproducible fashion

Methods

Results

Conclusion