Molekularno profiliranje karcinoma prijelaznog epitela gornjega dijela mokraćnoga sustava u nefropatiji aristolohične kiseline

doctoral thesis

• Title (english): Molecular profiling of the upper urinary tract transitional cell cancers associated with aristolochic acid nephropathy
• Author: Sandra Karanović
• Mentor: Bojan Jelaković (mentor)
• Mentor: Fran Borovečki (komentor)
• Committee member: nije dostupno (član povjerenstva)
• Granter: University of Zagreb; School of Medicine; Zagreb
• Defense date and country: 2015-10-28, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Clinical Medical Sciences
• Universal decimal classification (UDC): 61 - Medical sciences

Abstract

UVOD: Karcinomi prijelaznog epitela gornjeg dijela mokraćnog sustava (UUC) rijetki
su u općoj populaciji, ali se javljaju u gotovo 50% bolesnika s endemskom nefropatijom
(EN)/nefropatijom aristolohične kiseline (AAN). Aristolohična kiselina (AA) je
potvrđena kao etiološki čmbenik za razvoj EN/AAN i UUC. Cilj ovog istraživanja jest
odrediti obrasce genske ekspresije i mehanizme posttranskripcijske regulacije genske
ekspresije uključene u karcinogenezu uzrokovanu s AA.
MATERIJAL I METODE: Analizirani su parni uzorci tumora i priležećeg normalnog
tkiva od 13 bolesnika porijeklom iz hrvatskih i bosanskih endemskih područja.
Profiliranje miRNA učinjeno je pomoću RT-qPCR uređaja, dok je analiza genske
ekspresije učinjena na genskim čipovima. Imunohistokemijsko bojenje je učinjeno na
tkivnim microarrayima. Integrirana analiza podataka učinjena je koristeći više
bioinformatičkih programa.
REZULTATI: Identificirani su potpisi od 138 miRNA (74 povišene, 64 snižene) i 5438
(2021 povišene, 3417 snižene) značajno modulirane mRNA. Integrirajući mRNA i
miRNA podatke, detektirano je 1159 inverzno koreliranih mRNA, ciljnih molekula od
45 povišenih miRNA te 703 inverzno korelirane mRNA, ciljne molekule od 44 snižene
miRNA. Među najviše rangiranim dismoduliranim miRNA istaknule su se miR-200
obitelj, miR-17-92 skupina, let-7, miR-143/45 skupina i miR-23b. Opsežna biološka
analiza integriranih mRNA i miRNA podataka identificirala je sljedeće značajne
kategorije: stanični ciklus, odgovor na DNA oštećenje, popravak DNA, karcinom
mokraćnog mjehura, deregulacija onkogena, razvojni signalni putevi, remodeliranje
strukture kromatina. Podaci dobiveni molekularnim profiliranjem potvrđeni su
imunohistokemijski.
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ZAKLJUČCI: Tumorigeneza UUC uključuje procese posredovane djelovanjem
miRNA: rast tumora posljedično aktivaciji onkogena i represiji tumor supresor gena,
odnosno utišavanje kromatina i represiju prometastatskih programa uz posljedičnu
ograničenu metastatsku sposobnost ovog tipa tumora, što je u skladu s fenotipom ovih
tumora.

Abstract (english)

INTRODUCTION: Upper urinary tract urothelial cancers (UUC) are rare in general
population but occur in almost 50% of patients with endemic nephropathy
(EN)/aristolochic acid nephropathy (AAN). Aristolochic acid (AA) was proven as the
etiological factor both for EN/AAN and UUC. The goal of this study was to establish
patterns of gene expression and posttranscriptional gene regulation mechanisms
involved in AA induced carcinogenesis.
MATERIALS AND METHODS: Paired samples of tumors and adjacent normal
urothelial tissues of 13 patients from Croatian and Bosnian endemic regions were
analyzed. MiRNA profiling was performed by RT-qPCR, while mRNA expression
profiling was done using microarray technology. Immunohistochemistry was performed
on tissue microarrays. Integrative analysis of data was done using number of
bioinformatical programs.
RESULTS: A signature of 138 miRNAs (74 up, 64 down) and 5438 (2021 up, 3417
down) significantly modulated mRNAs were identified. Integrating miRNA and mRNA
data, 1159 predicted, inversely correlated targets of the 45 upregulated miRNAs and
703 predicted, inversely correlated targets of the 44 downregulated miRNAs were
identified. Top ranked differentially expressed miRNA included miR-200 family, miR-
17-92 cluster, let-7, miR-143/145 cluster, and miR-23b. Comprehensive biological
interpretation and mining of the integrated miRNA:mRNA data set identified pathways
of cell cycle, DNA damage response and DNA repair, bladder cancer, deregulated

oncogenic and developmental signaling pathways and remodeling of the chromatin
structure. Molecular profiling data were validated by means of immunohistochemistry.
CONCLUSIONS: UUC tumorigenesis appears to involve primarily miRNA-mediated
process of oncogenic activation of tumor growth and repression of tumor suppressor
genes, and chromatin silencing and repression of pro-metastasis programs rendering the
non-metastatic nature of these carcinomas, what is in line with phenotype of this
specific type of tumor.

• Keywords: medicina
• Keywords (english): medicine
• Language: croatian
• URN:NBN: urn:nbn:hr:105:632875
• Study programme: Title: Biomedicine and Health Sciences; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti, područje biomedicine i zdravstva (doktor/doktorica znanosti, područje biomedicine i zdravstva)
• Type of resource: Text
• Extent: 175 str.
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-05-05 07:37:01