Title Analiza ekspresije CTLA-4 u keratoakantomu
Title (english) Expression analysis of CTLA-4 in keratoacanthoma
Author Dora Knezović
Mentor Irena Drmić-Hofman (mentor)
Committee member Merica Glavina Durdov (predsjednik povjerenstva)
Committee member Neira Puizina Ivić (član povjerenstva)
Committee member Ivana Pavlinac Dodig (član povjerenstva)
Granter University of Split School of Medicine (Medical chemistry and biochemistry) Split
Defense date and country 2021, Croatia
Scientific / art field, discipline and subdiscipline BIOMEDICINE AND HEALTHCARE Basic Medical Sciences Medical Biochemistry
Abstract Ciljevi: Keratoakantom je keratinizirajući kožni tumor s centralnim kraterastim udubljenjem, nejasnog uzroka. Obilježavaju ga brzi rast i česta spontana regresija. Protein povezan s citotoksičnim limfocitima T ili CTLA-4 inhibicijski je receptor iz obitelji CD28 koji zaustavlja aktivaciju reagirajućih T-stanica te posreduje suprimirajuće funkcije regulacijskih T-stanica. Cilj studije je utvrditi izražaj CTLA-4 u centru, bazi keratoakantoma i zdravom lateralnom rubu kože kao kontroli te ispitati
... More ovisi li izražaj o stadiju keratoakantoma, spolu i dobi.
Materijali i metode: Istraživanje je obuhvatilo 30 uzoraka tkiva uklopljenog u parafinski blok, dobivenih nakon ekscizije keratoakantoma. U imunofluorescencijskom bojenju korišteno je anti–CTLA-4 zečje antitijelo (ab237712, Abcam, UK) te sekundarno antitijelo obilježeno fluoresceinom, a preparati su analizirani fluorescentnom mikroskopijom. Broj stanica pozitivnih na CTLA-4 izražen je u postotku u odnosu na ukupan broj stanica po vidnom polju (400 x).
Rezultati: Protein CTLA-4 izražen je u centru tumora u 28 (93.33%) slučajeva, na bazi tumora u 28 (93.33%) slučajeva, a značajno manje na zdravom lateralnom resekcijskom rubu, u 17 (56.67%) uzoraka. U centralnom dijelu tumora postoji statistički značajna razlika u izražaju CTLA-4 među stadijima keratoakantoma (P = 0.020). Postoji statistički značajna razlika između skupine s proliferativnnim stadijem i skupine s neodređenim stadijem keratoakantoma (P = 0.037) te između skupine s proliferativnim stadijem i skupine s regresijskim stadijem keratoakantoma (P = 0.022). Statistički značajna razlika prema spolu nije pronađena u ekspresiji CTLA-4 u centru tumora (P = 0.546), bazi tumora (P = 0.151), niti na zdravom lateralnom resekcijskom rubu (P = 0.914). Nije utvrđena statistički značajna razlika među dobnim skupinama s obzirom na ekspresiju CTLA-4 u centru tumora (P = 0.592), na bazi tumora (P = 0.888) te na zdravom resekcijskom rubu (P = 0.821).
Zaključci: Izražaj CTLA-4 je značajno veći u tumoru (centar i baza) nego na zdravom lateralnom resekcijskom rubu. Regresijski stadij keratoakantoma ima veći izražaj CTLA-4 nego proliferativni stadij. Spol i dob nisu povezani s izražajem CTLA-4. Za dodatna pojašnjenja uloge CTLA-4 u razvoju i/ili regresiji keratoakantoma, potrebna su daljnja istraživanja u smijeru fenotipizacije CTLA-4 pozitivnih stanica. Less
Abstract (english) Objectives: Keratoacanthoma is a dome-shaped skin tumor with a centralized keratinous plug, characterized by initial rapid growth and often spontaneous regression. Cytotoxic T-lymphocyte-associated protein 4 or CTLA-4 is an inhibitory receptor, belonging to the CD28 immunoglobulin subfamily, that inhibits T cell activation and mediates regulatory T-cell suppressive activity. The aim of this study was to determine whether there is a difference in CTLA-4 expression in the keratoacanthoma center,
... More at the base and in the healthy lateral resection margin as a control, as well as to determine whether it is dependent on keratoacanthoma stage, sex, and age.
Materials and methods: The study included 30 samples of paraffin-embedded tissue, obtained after KA excision. Anti–CTLA-4 rabbit antibody (ab237712, Abcam, UK) and a secondary antibody labeled with fluorescein were used for immunofluorescence staining. The slides were analyzed by fluorescence microscopy. The number of CTLA-4-positive cells was expressed as a percentage of the total number of cells per field of view (400 x).
Results: CTLA-4 protein was expressed in the tumor center in 28 (93.33%) cases, at the tumor base in 28 (93.33%) cases, and significantly less on the healthy lateral resection margin, in 17 (56.67%) samples. Statistically significant difference in CTLA-4 expression between the stages of KA was found in the central part of the tumor (P = 0.020). Also, statistically significant difference was found between the group with the proliferative stage and the group without exact stage of keratoacanthoma (P = 0.037), as well as between the group with the proliferative stage and the group with the regression stage of KA (P = 0.022). No statistically significant gender difference was found in CTLA-4 expression in the tumor center (P = 0.546), tumor base (P = 0.151), or at the healthy lateral resection margin (P = 0.914). No statistically significant difference was found between age groups with respect to CTLA-4 expression in the tumor center (P = 0.592), at the tumor base (P = 0.888), and at the healthy resection margin (P = 0.821).
Conclusions: CTLA-4 expression is significantly higher in the tumor (center and base) than on the healthy lateral resection margin. The regression stage of keratoacanthoma has a higher expression of CTLA-4 than the proliferative stage. Gender and age were not associated with CTLA-4 expression. To further clarify the role of CTLA-4 in the development and/or in the regression of keratoacanthoma, additional research is needed to determine the phenotype of CTLA-4 positive cells. Less
Keywords
Keratoakantom
CTLA-4 Antigen
Izražaj gena
Keywords (english)
Keratoacanthoma
CTLA-4 Antigen
Gene Expression
Language croatian
URN:NBN urn:nbn:hr:171:455687
Study programme Title: Medicine Study programme type: university Study level: integrated undergraduate and graduate Academic / professional title: doktor/doktorica medicine (doktor/doktorica medicine)
Type of resource Text
File origin Born digital
Access conditions Open access
Terms of use
Created on 2021-07-28 11:50:19