Vpliv inhibicije modificirane 6-fosfofrukto-1-kinaze na metabolizem rakastih celic.

Vodopivec, Tina (author), Legiša, Matic (mentor)

Rak je med vodilnimi vzroki smrti po vsem svetu. Pojav raka se povečuje zaradi rasti in staranja prebivalstva ter vse večje razširjenosti ugotovljenih dejavnikov tveganja, posledično pa se povečuje potreba po odkritju učinkovitejših zdravil proti raku. Obstajajo različni načini zdravljenja raka, v zadnjem desetletju je bil reprogramiran metabolizem rakavih celic obravnavan kot nov obetaven cilj za razvoj novih učinkovin. Preizkusili smo delovanje dveh inhibitornih molekul pridobljenih z in silico presejevanjem, ki ciljajo enega izmed glavnih regulatornih glikolitičnih encimov – fosfofrukto kinazo 1 (PFK-1), na metabolizem celic T limfoma linije Jurkat in celic seva kvasovk Sacharomyces cerevisiae z vstavljenim krajšim fragmentom za PFK-1, sfPFKM. Ugotovili smo, da dodatek obeh inhibitorjev vpliva na produkcijo laktata, razlika v koncentraciji v 72 urah je znašala 33,9 mg/l v primerjavi z netretiranimi celicami. Opazili smo tudi rahel citostatični vpliv inhbitorjev, ki je bil bolj opazen pri inhibitorju št. 3. Ugotovili smo tudi, da ima dodatek inhibitorja št. 3 in inhibitorja št. 9 statistično značilen učinek na bazalni privzem kisika (OCR) celic Jurkat in zniža stopnjo izvenceličnega zakisanja (ECAR). Dodatek inhibitorjev št. 3 in 9 omogoča boljšo rast seva sfPFKM S. cerevisiae na glukoznem viru ogljika zaradi nižje stopnje glikolize in izboljšanju redoks ravnotežja v celici. Pridobljeni podatki v naši raziskavi kažejo, da inhibitor št. 3 in 9 uspešno zavirajo metabolizem raka in jih je smiselno uporabiti v nadaljnjih raziskavah.

• Language: Slovenian
• Keywords: Jurkat, Saccharomyces cerevisiae, rak, 6-fosfofrukto-1-kinaza
• Work type: Master's thesis/paper
• Typology: 2.09 - Master's Thesis
• Organization: BF - Biotechnical Faculty
• Publisher: [T. Vodopivec]
• Year: 2019
• PID: 20.500.12556/RUL-112732
• UDC: 606:616-006.6:582.282.23:577.15(043.2)
• COBISS.SI-ID: 9341305
• Publication date in RUL: 09.11.2019

Secondary language

• Language: English
• Title: Influence of inhibition of modified 6-fosfofrukto-1-kinase on cancer metabolism

Abstract

Cancer is among the leading causes of death worldwide. The incidence of cancer is increasing and consequently the need to find an effective anticancer drug is also increasing. There are different ways to treat cancer, and over the last decade reprogrammed cancer cell metabolism has been seen as a new promising target for the development of new active substances. We tested the action of two inhibitory molecules obtained by in silico screening and targeting one of the major regulatory glycolytic enzymes - phosphofructo kinase 1 (PFK-1), on the metabolism of Jurkat T lymphoma cell line and Sacharomyces cerevisiae yeast cells with the inserted short fragment pf PFK-1 enzyme, sfPFKM. The addition of both inhibitors was found to affect lactate production and the difference in concentration at 72 hours was 33.9 mg/l compared with untreated cells. An effect on cell growth was also observed. Addition of inhibitor nr. 3 also showed cytostatic effects that were greater than those of inhibitor nr. 9. We also found that the addition of inhibitor nr. 3 and inhibitor nr. 9 effect on basal oxygen uptake (OCR) of Jurkat cells and reduces extracellular acidification rate (ECAR) with statistical significance. Supplement of inhibitors nr. 3 and 9 allow better growth of S. cerevisiae sfPFKM strain on glucose carbon source due to lower glycolysis rate and improved redox balance in the cell. The data obtained in our study indicate that inhibitors no. 3 and 9 effectively inhibit the metabolism of cancer and they show promise for further research.

• Keywords: Jurkat, Saccharomyces cerevisiae, cancer, 6-phosphofructo-1-kinase