Analiza učestalosti i tipova mutacija KRAS i BRAF gena u karcinomima kolorektuma u populaciji Srbije
Frequency and mutation type analysis of KRAS and BRAF genes in colorectal carcinomas in Serbian population
Author
Jakovljević, Ksenija V.
Mentor
Stamenković-Radak, Marina
Committee members
Janković, Radmila
Mališić, Emina

Metadata
Show full item recordAbstract
Uvod: Kolorektalni kancer (CRC) se po svojoj učestalosti i smrtnosti nalazi u samom
vrhu svih kancera kod oba pola, kako u svetu, tako i u Srbiji. Za ranu kancerogenezu
ovog maligniteta karakteristične su somatske genske promene KRAS i BRAF
protoonkogena. Mutacije ovih gena predstavljaju molekularne biomarkere odgovora na
ciljanu terapiju, dok su rezultati o njihovoj ulozi kao prognostičkih i prediktivnih
faktora kontradiktorni. Podaci o učestalosti i tipu pojedinačnih KRAS i BRAF mutacija
kod CRC-a u našoj zemlji su malobrojni, a njihova veza sa kliničko-histopatološkim
karakteristikama tumora, karakteristikama bolesnika i tokom bolesti nije dovoljno
ispitana.
Cilj: Utvrđivanje prisustva, učestalosti i tipa KRAS mutacija i specifične BRAF p.V600E
mutacije kod bolesnika sa CRC-om u populaciji Srbije i povezanost prisustva i tipa ovih
mutacija sa kliničko-histopatološkim karakteristikama tumora i karakteristikama
bolesnika. Takođe, ispitivanje navedenih mutacija kao prediktivnih biomark...era
adjuvantne i EGFR-ciljane terapije.
Materijal i metode: U radu je analizirano 188 parafinskih uzoraka karcinoma
kolorektuma. DNK je izolovana komercijalnim kitom za izolaciju genomske DNK iz
parafinskih tkiva (QIAamp® DNA FFPE Tissue kit, QIAGEN). Detekcija prisustva i
tipa mutacija kodona 12 i 13 KRAS gena urađena je pomoću dva dijagnostički validirana
testa bazirana na eseju lančane reakcije polimeraze (PCR) u realnom vremenu (DxS
TheraScreen K-RAS PCR kit, QIAGEN) i PCR reakciji praćenoj reverznom
hibridizacijom (KRAS StripAssayTM, ViennaLab Diagnostics). Detekcija p.V600E
mutacije u BRAF genu urađena je analizom krive topljenja (HRM) PCR produkta i
metodom direktnog sekvenciranja. Za statističku obradu podataka korišćeni su Fišerov
egzaktni, χ2 i Log-Rank testovi...
Introduction: Colorectal cancer (CRC) incidence and mortality rates are among the
highest in both sexes worldwide, as well as in Serbia. The early carcinogenesis of this
malignancy is characterized by somatic gene alterations of KRAS and BRAF
protooncogenes. Mutations of these genes represent molecular biomarkers of response
to targeted therapy, while their roles as prognostic and predictive factors are still
contradictory. There are few data about the frequency and types of single KRAS and
BRAF mutations in CRC in our country, and their correlation with
clinicohistopathological characteristics of tumor, characteristics of patients and the
course of disease is still to be established.
Aim: Determination of presence, frequency and types of KRAS mutations and specific
BRAF p.V600E mutation in CRC patients in Serbian population and correlation of
presence and types of these mutations with tumor clinicohistopathological
characteristics and patient characteristics. Also, the examination of ...specified mutations
as potential predictive biomarkers of adjuvant and EGFR-targeted therapy.
Material and methods: In this study 188 CRC paraffin samples were analyzed. DNA
was extracted using the commercial kit for genomic DNA isolation from paraffin tissues
(QIAamp® DNA FFPE Tissue kit, QIAGEN). Detection of presence and types of KRAS
codon 12 and 13 mutations was performed with two validated diagnostic tests based on
real-time polymerase chain reaction (PCR) assay (DxS TheraScreen K-RAS PCR kit,
QIAGEN) and PCR reaction followed by reverse hybridization (KRAS StripAssayTM,
ViennaLab Diagnostics). BRAF p.V600E mutation was assessed by high resolution
melting (HRM) analysis of PCR products and automatic DNA sequencing. Fisher exact,
χ2 and Log-Rank tests were used for statistical analysis...
Faculty:
Универзитет у Београду, Биолошки факултетDate:
09-07-2012Projects:
- Pharmacodynamic and pharmacogenomic research of new drugs in the treatment of solid tumors (RS-41026)