ANTIANGIOGENI POTENCIJAL SUSTAVNE HIPERTERMIJE, CISPLATINE I NANOKRISTALA RESVERATROLA U INHIBICIJI RASTA EHRLICHOVOG SOLIDNOG KARCINOMA U MIŠA

doctoral thesis

• Title (english): ANTIANGIOGENIC POTENTIAL OF SYSTEMIC HYPERTHERMIA, CISPLATIN AND RESVERATROL NANOCRYSTALS IN INHIBITING THE GROWTH OF ERLICH SOLID CARCINOMA IN MICE
• Author: Darko Kuĉan
• Mentor: Nada Oršolić (mentor)
• Mentor: Snježana Ramić (komentor)
• Committee member: Domagoj Đikić (predsjednik povjerenstva)
• Committee member: Romana Gračan (član povjerenstva)
• Committee member: Jelena Knežević (član povjerenstva)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2023, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology
• Universal decimal classification (UDC): 577 - Biochemistry. Molecular biology. Biophysics

Abstract

Prilagodba na hipoksiĉni mikrookoliš kritiĉna je za preţivljavanje tumora i širenje metastaza. Hipoksijom inducirani ĉimbenik-1α (HIF-1α) igra kljuĉnu ulogu u ovoj prilagodbi stimulirajući proizvodnju pro-angiogenih ĉimbenika i inducirajući enzime potrebne za anaerobni metabolizam. Inhibitori histon deaceti-laza (HDACI), ukljuĉujući resveratrol, inhibiraju ekspresiju HIF-1α i mogu doprinjeti inhibiciji rasta tu-mora kroz snaţni anti-angiogeni uĉinak. Stoga je cilj ovog rada bio istraţiti kemosenzitivni potencijal resveratrola i njegovih nanokristala (NK Res) u kombinaciji s cisplatinom na inhibiciju tumorske angioge-neze, rasta tumora, i polarizacije makrofaga u miševa nositelja solidnog oblika Ehrlichovog ascitnog tu-mora (EAT) izloţenih sustavnoj hipertermiji. Nakon injiciranja 1×106 EAT stanica supkutano (sc) u bedro Balb/c miševa, miševi su obraĊeni resveratrolom i/ili NK Res per os pet uzastopnih dana poĉevši od 2. dana nakon injekcije tumora te cisplatinom intraperitonealno (ip) u dozi od 2,5 mg/kg 10. i 12. dana i u dozi od 5 mg/kg 15. dana. Odmah potom, miševi su bili izloţeni sustavnoj hipertermiji 15 minuta na tem-peraturi od 41 °C. Razina histon deacetilaza (HDAC), HIF-1α, proteina toplinskog šoka-70 i -90 (HSP-70/90), krvoţilnog endotelnog ĉimbenika rasta (VEGF) i gustoća krvoţilja, metaloproteinaze-2 i -9 (MMP-2/9), inducibilna sintaza dušikovog oksida (iNOS) i arginaza odreĊeni su u tumorskom tkivu uz praćenje rasta i volumena tumora te preţivljavanja ţivotinja. Saţimajući dobivene podatke, resveratrol moţe pojaĉati antitumorski uĉinak cisplatine moduliranjem aktivnosti HDAC i razine HSP70/90, ĉime pridonosi izravnoj toksiĉnosti na tumorske stanice, dokidanju rezistencije i rasta tumora, te posljediĉnom povećanju preţivljenja miševa. Hipertermija pojaĉava lokalni protok krvi u tumoru povećavajući akumu-laciju cisplatine unutar tumorskog tkiva i remodulira mikrookruţenje tumora kroz aktivaciju M1 makrofa-ga i imunosnog sustava na stanice tumora. NK Res ima bolju dostupnost, topivost i zadrţavanje u specifiĉnom krvoţilju tumora povećavajući protutumoski i antiangiogeni uĉinak cisplatine pri fiziološkim uvjetima dok u hiprtermalnim uvjetima moţe neznatno smanjiti uĉinkovitost cisplatine kroz promjene tumorskog mikrookoliša i HIF-NO signalizacije.

Abstract (english)

Adaptation to hypoxic enviroment is crucial for tumor survival and metastatic spread. Hypoxia induced factor - 1α (HIF-1α) is a key factor in this adaptation by stimulating the production of proangiogenic factors and by inducing enzymes needed for anaerobic metabolism. Histone deacetylase inhibitors (HDACI), including resveratrol inhibit HIF-1α expression and can contribute to inhibiton of tumor growth through potent antiangiogenic effect. The purpuse of this research was to explore chemosensitive potential of resveratrol and its nanocrystal formulation (NK Res) in combination with cisplatin on tumor angiogenesis inhibition, tumor growth and macrophage polarisation in mice bearing solid form of Erlich ascites tumor (EAT) exposed to systemic hyperthermia. After subcutaneous (sc) injection of 1 × 106 EAT cells in thigh of Balb/c mice, mice were treated with resveratrol and/or NK Res per os during five consecutive days starting from the second day after tumor injection and cisplatin intraperitoneally (ip) at a dose of 2.5 mg/kg on the day 10 and 12 and at a dose of 5 mg/kg on the day 15. Right after, mice were exposed to systemic hyperthermia lasting for 15 min at a temperature of 41 °C. Level of histone deacetylases (HDAC), HIF-1α, heat shock protein-70 and -90 (HSP-70/90), vascular endothelial growth factor (VEGF) and vascular density, metalloproteinases-2 and 9 (MMP-2/9), inducible synthase of nitric oxide (iNOS) and arginase were determined in the tumor tissue with monitoring of tumor growth and volume and animal survival. Summarizing the obtained data, resveratrol can enhance the antitumor effect of cisplatin by modulating HDAC activity and the level of HSP70/90, thus contributing to direct toxicity on tumor cells, overcoming resistance and tumor growth, and consequently increasing the survival of mice. Hyperthermia enhances local blood flow in tumor and increases cisplatin accumulation of cisplatin within tumor tissue and remodulates tumor microenviroment through M1 macrophage activation and immune system on tumor cells. NK Res has better bioavailability, solubility and retention in specific tumor vasculature and thereby increases antitumor and antiangiogenic effect of cisplatin in physiological conditions, while in hyperthermic conditions it can slightly lower the effect of cisplatin through tumor microenviroment changes and HIF-NO signalisation.

• Keywords: Nanokristali resveratrola; sustavna hipertermija; cisplatina; angiogeneza
• Keywords (english): Resveratrol nanocrystal; systemic hyperthermia; cisplatin; angiogenesis
• Language: croatian
• URN:NBN: urn:nbn:hr:217:539705
• Promotion: 2023
• Study programme: Title: Doctoral study; Study programme type: university; Study level: postgraduate; Academic / professional title: doktor/doktorica znanosti u području prirodnih znanosti (doktor/doktorica znanosti u području prirodnih znanosti)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Open access
• Created on: 2023-11-22 13:25:22