Utvrđivanje prisustva TGF-ß1 metodama tekuće i tkivne biopsije kod pacijenata s kolorektalnim karcinomom

master's thesis

• Title (english): Determination of TGF-ß1 presence by liquid and tissue biopsy methods in patients with colorectal cancer
• Author: Tamara Bašić
• Mentor: Anita Somborac Bačura (mentor)
• Committee member: Anita Somborac Bačura (predsjednik povjerenstva)
• Committee member: Olga Gornik Kljaić (član povjerenstva)
• Committee member: Marija Grdić Rajković (član povjerenstva)
• Granter: University of Zagreb; Faculty of Pharmacy and Biochemistry; (Department of medical biochemistry and haematology); Zagreb
• Defense date and country: 2023-09-06, Croatia
• Scientific / art field, discipline and subdiscipline: BIOMEDICINE AND HEALTHCARE; Pharmacy; Medical Biochemistry

Abstract

Kolorektalni karcinom bolest je čija je prevalencija posljednjih godina u stalnom rastu. U početnom je stadiju bez
simptoma, zbog čega je rana dijagnostika ključna za poboljšanje prognoze bolesti te smanjenje smrtnosti. Zbog
invazivnosti i ograničenja kolonoskopije te tkivne biopsije kao postojećih dijagnostičkih metoda, kao inovacija u
području onkologije razvijena je tekuća biopsija. To je neinvazivna i ponovljiva metoda kojom je iz periferne krvi
pacijenata moguće izolirati egzosome porijeklom iz tumorskog tkiva. Egzosomi prenose molekule s potencijalom da
se koriste u praćenju bolesti, a jedna od njih je protein TGF-ß1. Mutacije u njegovoj signalizaciji utječu na staničnu
proliferaciju, progresiju tumora te razvoj metastaza. Cilj ovog rada bio je utvrditi prisutnost TGF-ß1 u egzosomima
pacijenata s kolorektalnim karcinomom dobivenima metodom tekuće biopsije te je usporediti s rezultatima
imunohistokemijske analize tumorskog tkiva dobivenog biopsijom. Metodom tekuće biopsije izolirani su egzosomi iz
uzoraka plazme pacijenata s kolorektalnim karcinomom. U uzorcima je određena koncentracija proteina
spektrofotometrijskom metodom s bicinkoniničnom kiselinom te je provedena Western blot analiza. Prisutnost TGFß1 dokazana je u svim uzorcima, a napravljena je i relativna kvantifikacija rezultata. Imunohistokemijskom analizom
tumorskog tkiva dobivenog biopsijom u svim je uzorcima također dokazana prisutnost TGF-ß1. Iz dobivenih se
rezultata može zaključiti da su rezultati dobiveni tekućom biopsijom istovjetni onima dobivenima tkivnom biopsijom.
Navedeno, uz potrebna daljnja istraživanja, otvara vrata tekućoj biopsiji da se uvede u kliničku praksu kao neinvazivna
metoda koja omogućuje kvalitetan probir, postavljanje dijagnoze te praćenje bolesti kod pacijenata s kolorektalnim
karcinomom.

Abstract (english)

Colorectal cancer is a disease whose prevalence has been steadily increasing in recent years. It is asymptomatic in
early stages, which is why early detection is essential for improving prognosis and reducing mortality. Due to the
invasiveness and limitations of colonoscopy and tissue biopsy as existing diagnostic methods, liquid biopsy was
developed as an innovation in the field of oncology. It is a non-invasive and reproducible method by which it is
possible to isolate exosomes from the patient's peripheral blood. Exosomes carry molecules originating from tumor
tissue with potential to be used in disease monitoring, one of them being TGF-ß1. Mutations in TGF-ß1 signaling path
affect cell proliferation, tumor progression and development of metastases. The aim of this thesis was to determine the
presence of TGF-ß1 in exosomes from patients with colorectal cancer obtained by liquid biopsy and to compare it with
the results of immunohistochemical analysis of tumor tissue obtained by biopsy. Using liquid biopsy, exosomes were
isolated from plasma samples of patients with colorectal cancer. The protein concentration in all samples was
determined by spectrophotometric method with bicinchoninic acid and Western blot analysis was performed. The
presence of TGF-ß1 was proven in all samples, with relative quantification of the obtained results.
Immunohistochemical analysis of tumor tissue obtained by biopsy also demonstrated the presence of TGF-ß1 in all
samples. In conclusion, results obtained by liquid biopsy are identical to those obtained by tissue biopsy. The above,
along with the necessary further research, allows liquid biopsy to be introduced into clinical practice as a non-invasive
method that enables high-quality screening, diagnosis and disease monitoring in patients with colorectal cancer.

• Keywords: kolorektalni karcinom; tekuća biopsija; egzosomi; TGF-ß1
• Keywords (english): colorectal cancer; liquid biopsy; exosomes; TGF-ß1
• Language: croatian
• URN:NBN: urn:nbn:hr:163:957422
• Study programme: Title: Medical biochemistry; Study programme type: university; Study level: integrated undergraduate and graduate; Academic / professional title: magistar/magistra medicinske biokemije (magistar/magistra medicinske biokemije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2024-01-18 14:41:33