Ispitivanje biohemijskih i patohistoloških markera značajnih za prognozu karcinoma bubrežnih ćelija
Examination of biochemical and pathohistological markers important for the prognosis of renal cell carcinoma
Doktorand
Radovanović, MilanMentor
Džamić, ZoranČlanovi komisije
Radojević-Škodrić, SanjaIsaković, Aleksandra
Pekmezović, Tatjana
Bančević, Vladimir
Metapodaci
Prikaz svih podataka o disertacijiSažetak
Karcinom bubrežnih ćelija (RCC) je najčešći maligni tumor bubrega kod odraslih. Javlja se u 85%-
90% svih malignih tumora bubrega kod odraslih. Najveća incidencija karcinoma bubrega se beleži
kod ljudi između 60-70 godine starosti, češće kod muškaraca (1,5:1), dok se u 7% slučajeva
dijagnostikuje kod osoba mlađih od 40 godina. Prema Vankuverskoj klasifikaciji iz 2004 postoje
tri glavna histološka podtipa karcinoma bubrežnih ćelija i to: svetloćelijski tip RCC (engl. clear cell
RCC - ccRCC), papilarni tip RCC (engl. papillary RCC - pRCC) i hromofobni RCC (engl. chromophobe
RCC - chRCC) pri čemu je najčešći tip ccRCC (70-85% svih RCC).
Veliki problem u terapijskom pristupu RCC jeste njegova rezistencija na hemioterapiju. Imajući u
vidu da je angiogeneza ključan faktor koji doprinosi napredovanju svih tumora pa i RCC, kao i da
apoptoza i autofagija mogu doprineti hemorezistenciji RCC smatrali smo značajnim upravo
ispitivanje patohistoloških i biomolekulanih faktora kao potencijalnih marke...ra RCC i
prognostičkih faktora u odnosu na tok bolesti i njen ishod. Takođe, ispitivanjem angiogeneze,
apoptoze i autofagije u različitim tipovima karcinoma bubrega bi se mogao poboljšati terapijski
pristup, a samim tim i prognoza pacijenata obolelih od karcinoma bubrega, naročito onih sa
metastatskom bolešću.
U okviru ovog istraživanja su izvedene dve studije na istom uzorku. Uzorak su činili pacijenti sa
postavljenom dijagnozom RCC (90 pacijenata) i pacijenti sa ccRCC (30 pacijenata) koji su hirurški
lečeni na Klinici za Urologiju Kliničkog Centra Srbije. Jedna studija je obuhvatila patohistološko
ispitivanje ekspresije p53, VEGF, survivina i beta katenina u različitim podtipovima RCC (90
pacijenata) i njihovu međusobnu korelaciju kao i njihovu korelaciju sa stadijumom, gradusom i
ishodom bolesti. Za potrebe ovog dela istraživanja je primenjena imunohistohemijska metoda
primenom metode tkivnog mikroniza (engl. tissue microarray - TMA). Druga studija je obuhvatila
ispitivanje ekspresije gena i proteina uključenih u kontrolu procesa apoptoze i autofagije u ccRCC
kao najčešćem tipu RCC (30 pacijenata). Za potrebe ovog segmenta istraživanja primenjene su
RT-qPCR metoda i imunoblot analiza.
Dobijeni rezultati su pokazali da je u tkivu RCC prisutna ekspresija p53, VEGF, survivina i beta
katenina i da ekspresija ovih proliferativnih markera korelira pozitivno sa stadijumom i gradusom
bolesti (izuzev beta katenina koji je pokazao negativnu korelaciju). Takođe je pokazano da između
analiziranih markera nema međusobne korelacije. Istovremeno, ekspresija p53, VEGF i survivina
pokazuje negativnu korelaciju sa dužim preživljavanjem dok ekspresija beta katenina pokazuje
pozitivnu korelaciju. U tkivu ccRCC pokazana je povećana ekspresija iRNK za p21, p27, p53
tumorsupresor gene, Bax i Bad antiapoptotskih gena dok je ekspresija tumor supresora PTEN bila
viša u peritumorskom tkivu. Ekspresija iRNK za markere autofagije p62, Atg4 i Uvrag je bila viša u
tumorskom tkivu ccRCC...
85% -90% of all malignant kidney tumors. The highest incidence of kidney cancer is recorded in
people between 60-70 years of age, more often in men (1.5: 1), while in 7% of cases it is
diagnosed in people younger than 40 years. According to the 2004 Vancouver classification, there
are three main histological subtypes of renal cell carcinoma: clear cell type RCC (ccRCC), papillary
type RCC (papillary RCC) and chromophobic RCC (chRCC). ). The most common type is ccRCC
which makes 70-85% of all RCCs.
A major problem with the RCC's therapeutic approach is its resistance to chemotherapy. Given
that angiogenesis is a key factor which contributes to the progression of all tumors, including
RCC, and that apoptosis and autophagy may contribute to RCC chemoresistance, we considered
as an important goal of this study to examine pathohistological and biomolecular factors as
potential markers of RCC and prognostic factors in relation to the course and outcome of the
disease. Also, the examination o...f angiogenesis, apoptosis and autophagy in different types of
kidney cancer could improve the therapeutic approach, and thus the prognosis of patients with
kidney cancer, especially those with metastatic disease.
Within this research, two studies were performed on the same sample. The sample included the
patients diagnosed with RCC (90 patients) and patients with ccRCC (30 patients) who were
surgically treated at the Clinic of Urology of the Clinical Center of Serbia. One study included
pathohistological examination of p53, VEGF, survivin, and beta catenin expression in different
RCC subtypes and their correlation as well as their correlation with the stage, grade, and disease
outcome (90 patients). In this part of the research, the immunohistochemical analysis including
the tissue microarray (TMA) method was performed. The other study included apoptosis and
autophagy related genes and proteins in the ccRCC as the most common type of RCC (30
patients). In this segment of the research RT-qPCR method and immunoblot analysis were used.
The obtained results demonstrate that the expression of p53, VEGF, survivin and beta catenin is
present in RCC tissue and that the expression of these proliferative markers correlates positively
with the stage and degree of the disease (except beta catenin which showed a negative
correlation). No correlation was found between the analyzed markers. At the same time, the
expression of p53, VEGF and survivin shows a negative correlation with longer survival while the
expression of beta catenin shows a positive correlation. In ccRCC tissue, increased mRNA
expression was shown for p21, p27, p53 tumor suppressor genes, Bax and Bad antiapoptotic
genes while PTEN tumor suppressor expression was higher in peritumoral tissue. mRNA
expression for autophagy markers p62, Atg4, and Uvrag was higher in ccRCC tumor tissue...