Vpliv elektrokemoterapije s cisplatinom ali bleomicinom in sunitinibom na celice humanega raka trebušne slinavke : magistrsko delo

Cerovšek, Anja (avtor), Čemažar, Maja (mentor), Omerzel, Maša (komentor), Serša, Gregor (član komisije za zagovor), Žnidaršič, Nada (član komisije za zagovor)

Elektrokemoterapija je način zdravljenja, ki uporablja elektroporacijo v kombinaciji s citostatikom, bodisi bleomicinom ali cisplatinom, za dosego lokalne kontrole tumorja. Bleomicin v celico vstopa s prenašalnimi proteini po endocitotski poti, cisplatin pa s pomočjo bakrovih kanalčkov in pasivno difuzijo. Ker je nabor prenašalnih proteinov precej majhen, je posledično tudi privzem citostatika v celico omejen. Elektrokemoterapija temelji na električnih pulzih, ki destabilizirajo celično membrano v tumorjih in tako se bistveno poveča vnos hidrofilnih citostatikov, ki imajo oviran transport skozi celično membrano. Bleomicin je protitumorsko zdravilo, ki prek tvorbe dvojnih prelomov ustavi celično delitev in povzroča smrt celic. Cisplatin pa uničujejo rakave celice tako, da poškodujejo DNA tako da z DNA tvori adukte, zavira sintezo in podvojevanje DNA in povzroči apoptotično smrt celic. Glavni cilj magistrske naloge je bil določiti občutljivost celic raka trebušne slinavke na elektrokemoterapijo s cisplatinom ali bleomicinom. Zanimalo nas je tudi, ali se citotoksični učinek poveča, če tretiramo celice raka trebušne slinavke s kombinacijo elektrokemoterapije s citostatikom in sunitinibom, saj tega do sedaj še nismo zasledili v literaturi. Sunitinib malat je molekula oksindola, zasnovana za selektivno interakcijo z znotrajceličnimi mesti ATP receptorjev kinaz kot so VEGFR1–3, PDGFR, KIT, FLT3 in CSF1R. Citotoksično delovanje kombinirane terapije smo preverili na humani celični liniji BxPC-3 raka trebušne slinavke. S testom PrestoBlueTM smo določili podvojitveni čas celične linije BxPC-3, ki je 48 ur (2 dni). Najbolj optimalno so rastle celice, ko smo jih nasadili 2000 / vdolbinico. Dokazali smo, da so celice BxPC-3 bolj občutljive na elektrokemoterapijo z bleomicinom kot na elektrokemoterapijo s cisplatinom. Kombinacija elektrokemoterapije z inhibitorjem tirozin kinaz sunitinibom na rakavih celicah trebušne slinavke, je imela večji citotoksični učinek le v primeru uporabe bleomicna. Kombinirana terapija elektrokemoterapije z 0,14 µM bleomicinom in sunitinibom v treh različnih koncentracijah (2,5, 5 in 7,5 µM) je imela sinergistični učinek. Glede na pozitivne in vitro rezultete, bi bilo smiselno tovrstno zdravljenje testirati tudi na tumorskem modelu trebušne slinavke in vivo.

• Jezik: Slovenski jezik
• Ključne besede: elektrokemoterapija, citostatiki, sunitinib, citotoksičnost
• Vrsta gradiva: Magistrsko delo/naloga
• Tipologija: 2.09 - Magistrsko delo
• Organizacija: BF - Biotehniška fakulteta
• Kraj izida: Ljubljana
• Založnik: [A. Cerovšek]
• Leto izida: 2022
• Št. strani: IV, 62 f.
• PID: 20.500.12556/RUL-134837
• UDK: 616(043.2)
• COBISS.SI-ID: 97115139
• Datum objave v RUL: 04.02.2022

CEROVŠEK, Anja, 2022, Vpliv elektrokemoterapije s cisplatinom ali bleomicinom in sunitinibom na celice humanega raka trebušne slinavke : magistrsko delo [na spletu]. Magistrsko delo. Ljubljana : A. Cerovšek. [Dostopano 3 april 2025]. Pridobljeno s: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=slv&id=134837

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Effect of electrochemotherapy with cisplatin or bleomycin and sunitinib on human pancreatic cancer cells : magistrski študij - 2. stopnja

Izvleček

Electrochemotherapy is a treatment option, using electroporation in combination with cytotoxic drugs, either bleomycin or cisplatin, to achieve local tumor control. Bleomycin enters the cell through endocytic transfer proteins and cisplatin through copper channels and passive diffusion. Because the set of transport proteins is quite small, consequently the uptake of the cytostatic into the cell is also limited.The set of transporter proteins is quite small, therefore the uptake of cytostatics into the cell is limited. Electrochemotherapy is based on electrical pulses that destabilize the cell membrane in tumors and thus significantly increase the cellular uptake of hydrophilic cytostatics that have obstructed transport across the cell membrane. Bleomycin is an antitumor drug that stops cell division through the formation of DNA double strand breaks leading to cell death. Cisplatin, on the other hand, destroy cancer cells by damaging DNA with formation of DNA adducts, consequently inhibiting DNA synthesis and replication, and causing apoptotic cell death. The aim of the master's thesis was to determine the sensitivity of pancreatic cancer cells to electrochemotherapy with cisplatin or bleomycin. We also investigated whether the cytotoxic effect is enhanced when pancreatic cancer cells are treated with combined electrochemotherapy and sunitinib, as this has not been described in the literature to date. Sunitinib malate is an oxindole molecule known to selectively interact with intracellular sites of ATP receptor kinases such as VEGFR1–3, PDGFR, KIT, FLT3, and CSF1R. The cytotoxic effect of combined therapy was examined on the human BxPC-3 pancreatic cancer cell line. The PrestoBlueTM assay was used to determine the doubling time of the BxPC-3 cell line, which was equal to 48 hours (2 days). Most optimal cell growth was observed when we plated 2000 cells per well. We demonstrated that BxPC-3 cells were more sensitive to electrochemotherapy with bleomycin than with cisplatin. Combination of electrochemotherapy with the tyrosine kinase inhibitor sunitinib on pancreatic cancer cells had more pronounced cytotoxic effect than electrochemotherapy alone, only when bleomycin was used. Combination therapy of electrochemotherapy with 0.14 µM bleomycin and sunitinib in three different concentrations (2.5, 5 in 7.5 µM) had a synergistic effect. Given the positive in vitro results, it would make sense to test this type of treatment on a tumor model of the pancreas in vivo.

• Ključne besede: electrochemotherapy, cytostatics, sunitinib, cytotoxicity