Title In vitro probir novih derivata fenantrena na tumorskim staničnim linijama Title (english) In vitro evaluation of new fenantren derivatives on tumor cell lines Author Danijel Selgrad Mentor Sandra Kraljević Pavelić (mentor) Committee member Mirela Sedić (predsjednik povjerenstva) Granter University of Rijeka Defense date and country 2020-06-22, Croatia Scientific / art field, BIOTECHNICAL SCIENCES Abstract Danas se maligna oboljenja liječe različitim pristupima koji uključuju kirurške postupke, zračenje i terapiju lijekovima. Terapija antitumorskim lijekovima je pristup koji se temelji na induciranju citotoksičnosti za tumorske stanice. Prirodni spojevi su tijekom povijesti bili važan izvor u razvoju lijekova. Ti su spojevi važne početne strukture za razvoj lijekova specifičnog djelovanja odnosno važne strukture za dobivanje sintetičkih aktivnih tvari. Iz kolekcije novosintetiziranih spojeva provode se ispitivanja, kao primjerice testiranja na staničnim linijama zloćudnih tumora u čovjeka u uvjetima in vitro ili visokoprotočna istraživanja. Fenantreni su klasa aromatskih metabolita, od kojih neki, posjeduju prilično jaku citotoksičnu aktivnost protiv staničnih linija raka, kao što su A549 (adenokarcinom bazalnog epitela alveola), SK-OV-3 (humani adenokarcinom jajnika), HL-60 (humana promijelocitična leukemija), HeLa (karcinom grlića maternice), MCF-7 (karcinom dojke). Temeljem prethodnih istraživanja različitih struktura fenantrena koje su pokazale biološku aktivnost, uključujući i onu protutumorsku, ovim radom se željela utvrditi citotoksičnost i antiproliferativna djelotvornost nove serije derivata fenantrena na tumorskim staničnim linijama pomoću metode MTT testa koja je služila za analizu proliferacije stanica in vitro, te uz pomoć metode Western blot za analizu ekspresije potencijalnih meta u tretiranim stanicama. Pokusi su rađeni na tumorskim staničnim linijama: A549 (adenokarcinom pluća), CFPAC-1 (stanice duktalnog adenokarcinoma gušterače), HeLa (karcinom grlića maternice), HepG2 (stanice hepatocelularnog karcinoma jetra), SW620 (stanice adenokarcinoma debelog crijeva, metastaze) te na jednoj staničnoj liniji netransformiranih stanica fibroblasta HFF-1. Dicijano supstituirani akrilonitrili 8 i 11 i njihovi ciklički analozi 15 i 17 su pokazali najjaču antiproliferativnu aktivnost, dok su derivati 8 i 11 također pokazali i selektivnost prema HeLa i HepG2 staničnim linijama. Razina ubikvitinirane forme HIF-1α statistički se povećala u stanicama Hela i HepG2 nakon tretmana derivatima 8 i 11. Protein MEK 1/2 imao je smanjenu ekspresiju u stanicama HeLa i HepG2 nakon tretmana derivatima 8 i 11. Zaključno, ovaj diplomski rad pokazuje potencijal testiranih derivata fenantrena 8 i 11 kao potencijalnih kandidata za daljnju optimizaciju u procesu razvoja lijekova.
Abstract (english) Today, malignancies are treated with a variety of approaches that include surgical procedures, radiation, and drug therapy. Antitumor drug therapy is the approach based on cytotoxicity induction in tumor cells. Natural compounds have been an important source in drug development throughout history. These compounds are important starting structures for development of specific drugs or important structures for production of synthetic active substances. From a collection of newly synthesized compounds, tests are carried out, such as testing for human malignant tumor cell lines in vitro or in high-throughput studies. Phenanthrenes are a class of aromatic metabolites, some of them bearing fairly strong cytotoxic activity against cancer cell lines, such as A549 (adenocarcinoma of the basal epithelium of the alveoli), SK-OV-3 (human ovarian adenocarcinoma), HL-60 (human promyelocytic leukemia), HeLa (cervical cancer), MCF-7 (breast cancer). Guided by previous studies of various phenanthrene structures that have shown biological activity, including antitumor ones, this work was aimed to study cytotoxicity and antiproliferative efficacy of a new series of phenanthrene derivatives on tumor cell lines using the MTT assay method used for cell proliferation assessment in vitro and Western blot for analysis of expression of potential targets in treated cells. The experiments were performed on tumor cell lines: A549 (lung adenocarcinoma), CFPAC-1 (pancreatic ductal adenocarcinoma cells), HeLa (cervical carcinoma), HepG2 (hepatocellular carcinoma cells), SW620 (colon adenocarcinoma cells), and on one cell line of untransformed fibroblast cells HFF-1. Dicyano substituted acrylonitriles 8 and 11 and their cyclic analogs 15 and 17 showed the strongest antiproliferative activity, while derivatives 8 and 11 also showed selectivity for HeLa and HepG2 cell lines. The level of ubiquitinated form HIF-1α was statistically increased in Hela and HepG2 cells after treatment with derivatives 8 and 11. MEK 1/2 protein had reduced expression in HeLa and HepG2 cells after treatment with derivatives 8 and 11. In conclusion, this thesis demonstrates the potential of tested phenanthrene derivatives 8 and 11 as potential candidates for further optimization in the drug development process.
Keywords
fenantreni
antiproliferativni učinak
protutumorski učinak
HIF-1α
MEK 1/2
Keywords (english)
phenanthrenes
antiproliferative effect
antitumor effect
HIF-1α
MEK 1/2
Language croatian URN:NBN urn:nbn:hr:193:016963 Study programme Title: Biotechnology in medicine Type of resource Text File origin Born digital Access conditions Open access Terms of use Created on 2020-07-06 13:01:08
