Učinak različitih makrocikličkih spojeva na proliferaciju stanica tumora in vitro

master's thesis

• Title (english): The effect of different macrocyclic compounds on tumor cell proliferation in vitro
• Author: Nataša Ilić
• Mentor: Marijeta Kralj (mentor)
• Mentor: Maja Matulić (sumentor)
• Committee member: Marijeta Kralj (predsjednik povjerenstva)
• Committee member: Maja Matulić (član povjerenstva)
• Committee member: Nada Oršolić (član povjerenstva)
• Committee member: Marina Cindrić (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2009, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Makrociklički spojevi, posebno krunasti eteri, od izuzetne su važnosti u kemiji i biologiji, ponajviše zahvaljujući mogućnosti selektivnog vezanja kationa. Krunasti spojevi imaju slična svojstva kao prirodni ionofori,no njihovo antitumorsko djelovanje tek se počelo istraživati. U ovom radu, istražili smo učinak 9 makrocikličkih spojeva na proliferaciju ljudskih tumorskih stanica HeLa, SW620, MCF-7, H460 i MiaPaCa-2 kao i mehanizam djelovanja najaktivnijih spojeva. Odredili smo proliferaciju stanica MTT testom, pri čemu su spojevi LET-610, LET-612, LET-646 i LET-613 pokazali značajno citostatičko i/ili citotoksično djelovanje. Za neke od testiranih spojeva prethodno je dokazana važnost veličine hidrofilne šupljine, tipa atoma donora i grupe koje prodiru u šupljinu na vezanje iona, te hidrofobnog prstena sa supstituentima za ugradnju u membranu. Ove strukturne karakteristike uspjeli smo povezati s njihovom antiproliferativnom aktivnošću u tumorskim stanicama. Kako bi rasvijetlili mehanizam djelovanja najaktivnijeg testiranog spoja LET-613 (IC50 ≈ 2 μM), analizirali smo njegov utjecaj na stanični ciklus metodom protočne citometrije. LET-613 je doveo do zastoja stanica raka gušterače MiaPaCa-2 u G1 fazi staničnog ciklusa, smanjenja broja stanica u S fazi i kasnije do povećanja broja apoptotskih (subG1) stanica. Annexin V testom, koji specifično detektira i kvantificira apoptotske stanice dodatno smo potvrdili da spoj LET-613 i prethodno testirani LET-329 aktiviraju apoptozu u stanicama raka. Rezultati podupiru pretpostavku da krunasti spojevi narušavaju prijenos iona, što dovodi do poremećaja u staničnom ciklusu i aktivacije apoptoze u tumorskim stanicama.

Abstract (english)

Macrocyclic compunds, especially crown ethers are of great importance in chemistry and biology mostly due to their ability to selectively bind cations. Crown compounds possess ionophoretic properties, but their antitumor activity has been underexplored. In this diploma thesis we studied the effect of 9 macrocyclic compounds on proliferation of human tumor cells HeLa, SW620, MCF-7, H460 and MiaPaCa-2 and evaluated the mechanism of action of the most active compounds. We used MTT assay to assess the proliferation rate of tumor cells. Compounds LET-610, LET-612, LET-646 and LET-613 showed significant cytostatic and/or cytotoxic activity. For some compounds it was previously demonstrated that the size of hydrophilic cavity, type of donor atoms and groups that penetrate into the cavity affect ion binding, whence hydrophobic ring with supstituents is important for membrane insertion. We showed that these structural characteristics correlate perfectly with the antiproliferative activity in tumor cells. In order to elucidate the mechanism of action of the most active tested compound LET-613 (IC50 ≈ 2 μM), we used flow cytometry for the cell cycle analysis. LET-613 induced G1 arrest of human pancreatic cancer cell MiaPaCa-2, along with S phase reduction and subsequent increase in subG1 phase (apoptotic cells). Activation of apoptosis in human cancer cells treated with compounds LET-613 and previously tested LET-329 was confirmed by performing Annexin V test that specifically detects and quantifies apoptotic cells. The results support the hypothesis that crown compounds disrupt ion transport that leads to the cell cycle perturbations and activation of apoptosis in tumor cells.

• Keywords: makrociklički spojevi; prijenos iona; stanični ciklus; apoptoza; rak
• Keywords (english): macrocyclic compounds; ion transport; cell cycle; apoptosis; cancer
• Language: croatian
• URN:NBN: urn:nbn:hr:217:422827
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: graduate; Academic / professional title: magistar/magistra molekularne biologije (magistar/magistra molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2018-03-12 09:35:01