| Abstract | Kolorektalni karcinom (CRC) zloćudni je tumor debelog i završnog crijeva koji prema statističkim podatcima
predstavlja jedan od vodećih uzroka smrti od malignih oboljenja u svijetu. Bolest se često dijagnosticira tek
u kasnijim stadijima, a rano otkrivanje značajno utječe na poboljšanje izgleda za uspješno izlječenje. Tekuća
biopsija omogućila je detekciju specifičnih tumorskih biljega na neinvazivan način, a kao najkorisniji biljeg
pokazala se slobodna cirkulirajuća DNA (ccfDNA). U pacijenata s malignim oboljenjem dolazi do
oslobađanja cirkulirajuće tumorske DNA s očuvanim genskim i epigenskim karakteristikama tumora što
omogućuje praćenje dinamike bolesti praćenjem koncentracije i molekularnog profila ccfDNA. Signalni put
JAK2/STAT3 ima značajan utjecaj na proliferaciju tumorskih stanica i na metastatski potencijal tumora i
korisna je terapijska meta u liječenju malignih oboljenja. S obzirom da je najčešća mutacija JAK2 V617F
karakterizirana hiperaktivacijom JAK2, a posljedično i STAT3, cilj ovoga rada bio je utvrditi prisutnost
navedene mutacije u uzorcima ccfDNA 31 ispitanika s dijagnozom CRC-a. Zbog niskih koncentracija
ccfDNA u uzorcima tijekom izrade rada korištena je metoda digitalne lančane reakcije polimerazom (dPCR)
na sustavu QuantStudio™ 3D Digital PCR System (Applied Biosystems, Foster City, Kalifornija, SAD).
Rezultati su očitani mjerenjem fluorescencijskih signala VIC™ i FAM™ s obilježenih sondi gdje je VIC™
vezana za sondu specifičnu za alel divljeg tipa, a FAM™ za mutirani alel. Obradom i pregledom rezultata u
računalnom programu QuantStudio™ 3D AnalysisSuite Cloud Software (Applied Biosystems, Foster City,
Kalifornija, SAD) utvrđeno je da mutacija JAK2 V617F u uzorcima ccfDNA ispitanika s dijagnozom CRCa
nije prisutna. |
| Abstract (english) | Colorectal cancer (CRC) is a malignant tumour of the colon and rectum that, according to statistical data, is
one of the leading causes of cancer-related deaths worldwide. The disease is often diagnosed at later stages,
while early detection significantly improves the chances of successful treatment. Liquid biopsy has enabled
the detection of specific tumour markers in a non-invasive manner, with cell-free circulating DNA (ccfDNA)
emerging as the most useful marker. In patients with malignancies, circulating tumour DNA is released,
maintaining the genetic and epigenetic characteristics of the tumour, which allows for disease monitoring by
tracking the concentration and molecular profile of ccfDNA. The JAK2/STAT3 signaling pathway has a
significant impact on the tumour cell proliferation and the metastatic potential of the tumour, making it a
valuable therapeutic target in the treatment of malignancies. Since the most common mutation, JAK2 V617F,
is characterized by hyperactivation of JAK2 and, consequently, STAT3, the aim of this study was to
determine the presence of this mutation in ccfDNA samples from 31 CRC patient. Due to the low
concentrations of ccfDNA in the samples, digital polymerase chain reaction (dPCR) was used, performed on
the QuantStudio™ 3D Digital PCR System (Applied Biosystems, Foster City, California, USA). The results
were analysed by measuring the fluorescence signals of VIC™ and FAM™ dyes bound to the labeled probes,
where VIC™ is bound to the probe specific for the wild-type allele and FAM™ to the probe specific for the
mutated allele. The results were processed and analysed using the QuantStudio™ 3D AnalysisSuite Cloud
Software (Applied Biosystems, Foster City, California, USA) and it has been concluded that the JAK2 V617F
mutation is not present in the ccfDNA samples of the CRC patients. |
