Povezava epigenetskih in ekspresijskih markerjev pri raku glave in vratu

diplomsko delo univerzitetnega študijskega programa I. stopnje

Monika Ostroško (Author), Uroš Potočnik (Mentor), Bogdan Čizmarevič (Co-mentor), Helena Sabina Čelešnik (Co-mentor)

Abstract

Rak glave in vratu predstavlja obsežno skupino bolezni, v katero uvrščamo raka ustnic, ustne in nosne votline, žrela, grla, vratu in nekatere druge. Med temi tumorji opazimo velike razlike, ne le zaradi mesta izvora, marveč prihaja tudi do različnih simptomov bolezni, hitrosti širjenja, različni so načini zdravljenja ter izidi bolezni. V pomoč pri postavljanju diagnoze, spremljanju obnašanja tumorja in pravilnem izboru zdravljenja se uporabljajo biološki označevalci. V telesu prisotnost teh snovi, ki jih ustvari tumor ali zdrave celice kot reakcijo na tumor, odkrijemo s pomočjo laboratorijske preiskave bioloških vzorcev. V diplomskem delu smo želeli izmeriti ekspresijo genov DAPK1 in MMP9 v zdravih in bolnih tkivih ter vzpostaviti protokole za metilacijsko analizo gena DAPK1. Genska ekspresija in metilacija sta povezana biološka procesa, saj metilacija vpliva na vezavo specifičnih transkripcijskih faktorjev na DNA, ti pa uravnavajo izražanje genov. Pri raziskovanju nas je zanimalo, če bi lahko DAPK1 in MMP9 služila kot možna biološka označevalca pri bolnikih z rakom glave in vratu. Za ugotavljalnje izražanja teh genov v zdravih in bolnih tkivih smo uporabili metodo qRT-PCR. Za analizo metilacijskih vzorcev na promotorju gena DAPK1 smo testirali dve občutljivi in specifični tehniki za določanje metilacije DNA, metodo MS-HRM ter metodo COBRA. Za analizo metilacije promotorja DAPK1 pri 15 bolnikih smo nato uporabili metodo COBRA. Rezultati genske ekspresije pri 47 bolnikih z rakom glave in vratu so pokazali statistično signifikantno znižanje DAPK1 v rakavih tkivih v primerjavi z zdravimi tkivi istih pacientov, medtem ko rezultati MMP9 kažejo na statistično signifikantno povečanje ekspresije v rakavih tkivih. Z merjenjem genske ekspresije smo prišli do zaključka, da sta gena DAPK1 in MMP9 zanimiva kot potencialna biološka markerja pri raku glave in vratu. Zaradi majhnega števila bolnikov, ki so bili vključeni v metilacijsko analizo, nismo mogli postaviti zaključkov o metiliranosti promotorja DAPK1. Za jasne zaključke bodo potrebne nadaljne analize.

Keywords

rak glave in vratu;biološki označevalci;genska ekspresija;metilacija;diplomske naloge;

Data

Language:	Slovenian
Year of publishing:	2020
Typology:	2.11 - Undergraduate Thesis
Organization:	UM FKKT - Faculty of Chemistry and Chemical Engineering
Publisher:	[M. Ostroško]
UDC:	616-006.3(043.2)
COBISS:	34223363
Views:	457
Downloads:	66
Average score:	0 (0 votes)
Metadata:

Other data

Secondary language:	English
Secondary title:	Correlation of epigenetic and gene expression markers associated with head and neck cancer
Secondary abstract:	Head and neck cancer represents a spectrum of diseases, which includes cancer of the lips, oral and nasal cavities, throat, larynx, neck etc. There are significant differences between these tumors, including different symptoms, chance of spreading, different treatment requirements and different outcomes. To help diagnose and monitor these tumors and to help select proper treatments, biomarkers are used. In the body, the presence of these molecules, which are produced either by the tumor or by healthy cells in response to the tumor, is detected by laboratory testing of biological samples. In this work we wanted to measure the expression of DAPK1 and MMP9 genes in cancerous and adjacent healthy tissues and to set up a protocol for methylation analysis of the DAPK1 promoter. Gene expression and methylation are interconnected biological process, since methylation influences the binding of specific transcription factors to DNA, thereby interfering with their function in gene expression control. In this study, we wanted to investigate whether DAPK1 and MMP9 could serve as possible biomarkers in patients with head and neck cancer. To analyze gene expression in cancerous and healthy tissues, qRT-PCR was used. For methylation analysis of DAPK1 promoter, we tested two sensitive techniques for determining DNA methylation, MS-HRM and COBRA. For DAPK1 methylation analysis of biological samples from 15 patients, the COBRA technique was selected. The results of gene expression for 47 patients with head and neck cancer revealed a statistically significant decrease in DAPK1 espression in cancerous versus healthy tissues, while the results for MMP9 showed a statistically significant increase in MMP9 expression in cancerous tissues. The results of gene expression analysis suggest that DAPK1 and MMP9 could be of interest as potential biological markers in head and neck cancer. Due to the small number of patients included in the methylation analysis, we were not able to draw conclusions about the methylation of the DAPK1 promoter. Thus, further analyses will be needed for definitive answers.
Secondary keywords:	head and neck cancer;biomarkers;DAPK1;MMP9;gene expression;methylation;
Type (COBISS):	Bachelor thesis/paper
Thesis comment:	Univ. v Mariboru, Fak. za kemijo in kemijsko tehnologijo
Pages:	IX, 37 str.
ID:	11991857