Naslov (srp)

Антитуморски ефекти активних принципа изолованих из Onosma visianii на леукемијским лимфоцитима

Autor

Todorović, Željko, 1987-

Doprinosi

Đurđević, Predrag, 1971-
Milovanović, Marija, 1979-
Radosavljević, Gordana, 1976-
Vuković, Nenad, 1976-
Tarabar, Olivera, 1960-

Opis (eng)

Introduction: Chronic lymphocytic leukemia is a chronic lymphoproliferative diseasecharacterized by the accumulation of morphologically mature but immune immature Blymphocytes in the blood, bone marrow and lymphatic tissues. It occurs as a consequence ofclonal proliferation of malignantly transformed B lymphocytes and their resistance to apoptosis.Unlike chronic lymphocytic leukemia, which is the most common leukemia in the Westernworld, B prolymphocytic leukemia is a rare chronic lymphoproliferative disease characterized bypoor prognosis and significantly shorter survival compared to chronic lymphocytic leukemia.Both diseases, despite modern therapy, are still incurable.Shikonin and shikonin derivatives are naphthaquinone compounds that have shown significantantitumor activity in numerous studies. The mechanism of antitumor action of these substances isdiverse and includes inhibition of several signaling pathways, including STAT3, induction ofendoplasmic reticulum stress, hyperproduction of oxygen free radicals and many others. The finalresult is apoptosis of the treated cells.Aim: The main aim of this study was to examine the antitumor activity of two shikoninderivatives derived from the plant Onosma visianii, isobutyrylshikonin and αmethylbutyrylshikonin in BCL1, mouse CLL cells and JVM-13, human B-PLL cellsMaterial and methods: The cytotoxicity of shikonin derivatives was measured by an MTT test.Cell death, proliferation, cell cycle, and expression of molecules that control these processes wereanalyzed by flow cytometry. Expression of STAT3-regulated genes was analyzed by real-time qRT-PCR (Quantitative Real-Time Polymerase Chain Reaction). The antitumor effects of shikoninderivatives in vivo were analyzed, using flow cytometry, by detection of leukemia cells in theperipheral blood and spleens of mice intravenously injected with BCL1 cells.Results: Isobutyrylshikonin and α-methylbutyrylshikonin induced cell cycle disturbances andapoptosis, inhibited proliferation, and decreased expression of phospho-STAT3 and downstreamregulated molecules in BCL1 and JVM-13 cells. IBS and MBS decreased the percentage ofleukemia cells in vivo. The link between the decrease in phosphorylated STAT3 by MBS andIBS and BCL1 cell death was confirmed by detection of enhanced cell death after addition ofAG490, an inhibitor of Jak2 kinase.Conclusion: It seems that IBS and MBS, by decreasing STAT3 phosphorylation, triggerapoptosis, inhibit cell proliferation, and attenuate leukemia cell stemness. Results suggest thatisobutyrylshikonin and α-methylbutyrylshikonin, with additional preclinical studies, may bepotential therapies for chronic lymphocytic and B prolymphocytic leukemia.

Opis (srp)

Uvod: Hronična limfocitna leukemija je hronično limfoproliferativno oboljenje kojekarateriše nakupljanje morfološki zrelih ali imunski nezrelih B limfocita u krvi,kostnoj srži i limfnim tkivima. Nastaje kao posledica klonske proliferacije malignotransformisanih B limfocita i njihove rezistencije na apoptozu. Za razliku odhronične limfocitne leukemije koja je najčešća leukemija u zapadnom svetu, Bprolimfocitna leukemija je retka hronična limfoproliferativna bolest kojukarateriše loša prognoza i značajno kraće preživaljavanje u poređenju sa hroničnomlimfocitnom leukemijom. Obe bolesti su, i pored savremene terapije, i dalje neizlečive.Shikonin i derivati shikonin-a su naftohinonska jedinjenja koja su u brojnimistraživanjima pokazala značajno antitumorsko dejstvo. Mehanizam antitumorskogdejstva ovih supstanci je raznolik i uključuje inhbiciju više signalnih puteva međukojima je i STAT3.Cilj ovog istraživanja je ispitivanje potencijalnog antitumorskog dejstvo dva derivatašikonina, dobijenih iz biljke Onosma visianii, isobutyrylshikonin-a i αmethylbutyrylshikonin-a, in vitro korišćenjem BCL1 ćelijske linije mišjeleukemije/limfoma i JVM-13 ćelijske linije humane B-prolimfocitne leukemije kao i invivo korišćenjem eksperimentalnog modela hronične limfocitne leukemije.Materijal i metode: Za ispitivanje citotoksičkog efekta derivata šikonina korišćenje MTT test. Tip ćelijske smrti, proliferacija, ćelijski ciklus i ekspresija molekulakoji učestvuju u kontroli ovih procesa analizirani su protočnom citometrijom.Ekspresija gena koji su regulisani STAT3 molekulom ispitana je q-RT-PCR (od eng.Quantitative Real-Time Polymerase Chain Reaction) metodom. Antitumorski efekti derivatašikonina in vivo ispitani su detekcijom leukemijskih ćelija u perifernoj krvi islezini miševa kojima su prethodno intravenski injektovane BCL1 ćelija protočnomcitometrijom.Rezultati: Isobutyrylshikonin i α-methylbutyrylshikonin izazivaju poremećaje ćelijskogciklusa i indukuju apoptozu, inhibiraju proliferaciju i smanjuju ekspresijufosforilisane forme STAT3 i gena čiju ekspresiju kontroliše ovaj transkripcionifaktor u BCL1 i JVM-13 ćelijama. Ispitivani derivati šikonina smanjuju procenatleukemijskih ćelija u perifernoj krvi i slezini miševa sa CLL. Veza između smanjenjaekspresije fosforilisanog STAT3 u ćelijama tretiranim ispitivanim derivatimašikonina i smrti BCL1 ćelija potvrđena je detekcijom pojačane smrti ćelija tretiranihi inhibitorom Jak2 kinaze.Zaključak: Isobutyrylshikonin i α-methylbutyrylshikonin najverovatnije smanjenjemfosforilacije STAT3 indukuju apoptozu, inhibiraju ćelijsku proliferaciju iatenuiraju matičnost lekuemijskih ćelija. Ovi rezultati ukazuju da isobutyrylshikonin i αmethylbutyrylshikonin, uz dodatna pretklinička ispitivanja, mogu biti potencijalnaterapija za hroničnu limfocitnu i B prolimfocitnu leukemiju.

Opis (srp)

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Jezik

srpski

Datum

2021

Licenca

Ovo delo je licencirano pod uslovima licence
Creative Commons CC BY 3.0 AT - Creative Commons Autorstvo 3.0 Austria License.

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