Title Prognostička vrijednost imunohistokemijske izraženosti fibronektina i galektina-3 u urotelnom karcinomu mokraćnoga mjehura Title (english) Prognostic value of immunohistochemical expression of fibronectin and galectin-3 in urothelial carcinoma of the urinary bladder Author Šoip Šoipi 305765950 Mentor Majda Vučić (mentor)Mentor Borislav Spajić (komentor)Committee member Hrvoje Čupić (predsjednik povjerenstva)Committee member Božo Krušlin (član povjerenstva)Committee member Željko Kaštelan (član povjerenstva)Committee member Sven Seiwerth (član povjerenstva)Committee member Zoran Peršec (član povjerenstva)Committee member Vlaho Brailo (član povjerenstva)Granter University of Zagreb Defense date and country 2020-10-09, Croatia Scientific / art field, BIOMEDICINE AND HEALTHCARE Universal decimal classificationUDC ) 616 - Pathology. Clinical medicine Abstract Rak mokraćnog mjehura zahtjeva učestale višegodišnje kontrolne preglede (cistoskopija i citologija urina) zbog svoje sklonosti nastanku recidiva ili progresije u viši stadij bolesti. Na mikroskopskoj razini tumor se sastoji od epitelnih stanica, ali i strome koja pruža funkcionalnu potporu. Stanice strome i vanstanični matriks (ECM) podložni su promjenama važnim za širenje karcinoma i agresivno ponašanje. Galektin-3 i fibronektin proteini su ECM-a koji imaju ulogu u stvaranju veza između stanica. U ovom istraživanju određivana je izraženost fibronektina i galektina-3 u stromi i epitelu urotelnog karcinoma te korelacija s patohistološkim parametrima prognoze (TNM i gradus) i kliničkim parametrima (dob, spol, pojava recidiva, učestalost recidiva, vremenski razmak između pojave recidiva). U istraživanju je korišten arhivski materijal dobiven nakon transureteralne resekcije te cistektomije i limfadenektomije. Analizirani su uzorci 110 pacijenata, podjeljeni u skupine; karcinomi: Ta/T1 niskog gradusa, Ta/T1 visokog gradusa, T2N0 te T2N1/2. U imunohistokemijskoj analizi korištena su monoklonska protutijela na fibronektin i galektin-3.Statističkom analizom rezultata ustanovljena je statistički značajna pozitivna korelacija izraženosti fibronektina u epitelnoj komponenti tumora s T-stadijem (p=0,001) i N- stadijem bolesti (p=0,037) te negativna korelacija bojenja stromalne komponente s T stadijem bolesti (p=0,001). Ustanovljena je i pozitivna korelacija izraženosti galektina-3 u epitelu tumora i T stadija bolesti (p=0,046), što znači da viša ekspresija obilježava agresivniji tumor, a u metastatskim tumorima u limfne čvorove brži povrat bolesti (p=0,00). Stoga možemo zaključiti da s porastom ekspresije fibronektina u epitelnim tumorskim stanicama i smanjenjem ekspresije u stromi tumori pokazuju agresivniji obrazac rasta obzirom da s rastom ekspresije fibronektina u epitelu tumori pokazuju viši gradus tj. slabije su diferencirani te pokazuju sklonost infiltrativnom rastu i metastaziranju u limfne čvorove. U kliničkoj praksi, temeljem ovog istraživanja navedeni proteini mogli bi se koristiti kao jedan od čimbenika probira za tumore koji će pokazivati sklonost agresivnijem biološkom ponašanju, a što bi moglo dovesti do izdvajanja skupine bolesnika u ranim stadijima bolesti, a kojima su potrebne češće kontrole i eventualna dodatna terapija.
Abstract (english) INTRODUCTION Carcinoma of the bladder is the second most common malignancy of urological organs and is still a worldwide health problem. Urothelial (transitional cell) carcinoma is the predominant histologic type. The spectrum of bladder cancer includes non-muscle-invasive (superficial), muscle-invasive, and metastatic disease, each with its own clinical behavior, biology and prognosis. Carcinoma of the bladder is at high risk of recurrence and progression of disease. Microscopically the tumor is composed of epithelial cells and stroma. The stroma consists of fibroblasts/myofibroblasts, fat, immune, vascular and smooth muscle, cells along with the extracellular matrix (ECM) and extracellular molecules. While none of these cells are themselves malignant, due to their environment, their interactions with each other, and directly or indirectly with the cancer cells, they acquire an abnormal phenotype and altered function. Extracellular matrix molecules in the urinary carcinoma of the bladder seem to play an important role in the development of invasion, progression and metastasis. Galectin-3 and fibronectin are ECM proteins that play an important role in cell adhesion. AIM The aim of this research was to analyse immunohistochemical expression of fibronectin and galectin-3 in epithelial and stromal cells of urothelial bladder carcinoma and correlate it with TNM stage, tumor grade and clinical parameters (age, sex, recurrence rate). PATIENTS AND MATERIALS Archived material from transurethral resection, cystectomy and lymphadenectomy was used in this research. Specimens carcinomas were analysed from 110 patients and divided into four groups; Ta/T1 low grade, Ta/T1 high grade, T2N0 stage and T2N1/2. Specimens where fixed in 10% buffered formalin, cut at 5μm thickness and routinely stained with hematoxylin and eosin method for diagnosis and TNM and grade classification. Additional immunohistochemistry was performed by monoclonal antibodies to fibronectin (Novocastra Laboratories 1:400) and galectin-3 (Novocastra Laboratories 1:100). The intensity of Šoip Šoipi, Disertacija immunostaining was graded semiquantitative on a scale of 0-3 and expressed as 0 = 0%; 1 = up to 33%; 2 = more than 33% to 66%; and 3 = more than 66% of positive epithelial and stromal cells. RESULTS There was statistically significant positive correlation in expression of fibronectin in epithelial tumor cells with T (p=0.001) and N tumor stage (p=0.037) as well as negative correlation in expression of fibronectin in stromal component with T tumor stage (p=0.001). There was statistically significant positive correlation in expression of galectin-3 in epithelial tumor cells compared with T tumor stage (p=0.046) connected with the more aggressive tumor biology and positive correlation in expression of galectin-3 in tumor with metastases in lymph nodes with disease relapse rate (p=0.00). CONCLUSION Increased fibronectin expression in epithelial tumor cells and decreased fibronectin in stromal tumor component was related to more aggressive tumor biology, higher tumor grade and more infiltrative tumor growth with metastatic spread to lymph nodes. These observations suggest a role for fibronectin and galectin-3 as biomarkers for bladder cancer diagnostics, staging, outcome and prognosis. In clinical practice these molecular markers could be used in creating subgroups of patients that would benefit from more frequent follow up or additional therapeutic approaches.
Keywords
tumori mokraćnog mjehura
karcinom prijelaznog epitela
biološki biljezi
Keywords (english)
Urinary Bladder Neoplasms
Carcinoma
Transitional Cell
Biomerkers
Language croatian URN:NBN urn:nbn:hr:127:422141 Promotion 2020 Study programme Title: dental medicine Type of resource Text Extent 86 str; 29,7 cm File origin Born digital Access conditions Open access Terms of use Created on 2020-11-06 11:12:51
