o:29790 Interakcija mitogenom aktiviranih protein kinaza i autofagije u diferencijaciji HL-60 leukemijskih ćelija u makrofage forbol-12-miristat-13-acetatom doktorska disertacija Interaction of mitogen-activated protein kinases and autophagy in macrophage differentiation of HL-60 leukemia cells by phorbol-12-myristate-13-acetate : doctoral dissertation sr Forbol-12-miristat-13-acetat (PMA), aktivator protein kinaze C (PKC), indukuje diferencijacijućelija akutne mijeloidne leukemije (AML) u makrofage. Pokazano je da autofagija učestvuje udiferencijaciji ćelija hematopoeze i leukemijskih ćelija. Mitogenom aktivirane protein kinaze(MAPK) mogu da aktiviraju autofagiju i diferencijaciju ćelija. Cilj ove disertacije je da se ispitauloga autofagije i MAPK u diferencijaciji HL-60 leukemijskih ćelija u makrofage pod dejstvomPMA, kao i uloga MAPK u indukciji autofagije. PMA je indukovala diferencijaciju HL-60 ćelija umakrofage, što je pokazano povećanom ekspresijom makrofagnih markera CD11b, CD13, CD14,CD45, EGR1, CSF1R i IL-8. Prisustvo autofagije u diferencijaciji HL-60 ćelija pokazano јеdetekcijom zakišeljavanja citoplazme, povećanjem autofagnog fluksa, povećanjem ekspresije ATGgena i oslobađanjem beklina-1 iz kompleksa sa Bcl-2. Autofagija je aktivirana izlaskom autofagnogrepresora ZKSCAN3 iz jedra i ulaskom aktivatora autofagije FOXO1/3 i TFEB u jedro HL-60ćelija. Aktivacija autofagije je nezavisna od AMPK i Akt/mTORC1. Međutim, PMA je aktivacijomPKC dovela do stimulacije ERK i JNK, što je povećalo konverziju LC3-I u LC3-II. Farmakološka igenska inhibicija ERK ili JNK je smanjila konverziju LC3, ekspresiju ATG gena, premeštanjeFOXO1/3 i TFEB u jedro, disocijaciju beklina-1 iz kompleksa sa Bcl-2 i ekspresiju makrofagnihmarkera CD11b, CD45, CSF1R, EGR1 i IL-8. Farmakološka i genska inhibicija autofagije jesprečila diferencijaciju HL-60 ćelija u makrofage. Ovi rezultati ukazuju da ERK i JNK učestvuju udiferencijaciji HL-60 ćelija u makrofage pod dejstvom PMA, aktivacijom autofagije, što predstavljapotencijalnu metu za poboljšanje diferencijacione terapije AML. Protein kinase C (PKC) activator phorbol-12-myristate-13-acetate (PMA) induces macrophagedifferentiation of acute myeloid leukemia (AML) cells. Autophagy has been shown to participate inthe differentiation of hematopoietic and leukemic cells. It is well known that mitogen-activatedprotein kinases (MAPK) can activate autophagy and cell differentiation. The aim of this thesis is toexamine the role of autophagy and MAPK activation in PMA-induced macrophage differentiationof HL-60 leukemia cells as well as the role of MAPK in the induction of autophagy. PMA inducedmacrophage differentiation of HL-60 cells, as shown by increased expression of macrophagemarkers CD11b, CD13, CD14, CD45, CSF1R, EGR1 and IL-8. The activation of autophagy duringthe differentiation of HL-60 cells has been demonstrated by the increase of cytoplasmicacidification, augmented autophagic flux, increased ATG gene expression, and the release of beclin-1 from the complex with Bcl-2. Autophagy in HL-60 cells has been activated by the cytoplasmictranslocation of the autophagy repressor ZKSCAN3 and nuclear translocation of autophagyactivators FOXO1/3 and TFEB. Stimulation of autophagy was independent of AMPK andAkt/mTORC1 activation. However, PMA activation of PKC stimulated ERK and JNK andincreased the LC3 conversion. Pharmacological and genetic inhibition of ERK or JNK reduced LC3conversion, ATG gene expression, FOXO1/3 and TFEB nuclear translocation and expression ofmacrophage markers CD11b, CD45, CSF1R, EGR1 and IL-8. Inhibition of autophagy suppresseddifferentiation of HL-60 leukemia cells induced by PMA. These results indicate that ERK and JNK-dependent autophagy participates in PMA-induced differentiation of HL-60 cells into macrophages,which represents a potential target for improving AML differentiation therapy. Medicina - Molekularna medicina / Medicine- Molecular medicine Datum odbrane: 01.03.2023. akutna mijeloidna leukemija, forbol-12-miristat-13-acetat, diferencijacija, autofagija, beklin-1, ERK, JNK acute myeloid leukemia, phorbol 12-myristate 13-acetate, differentiation, autophagy, beclin-1, ERK, JNK 616.155:577.2(043.3) 91552100 115220233 91552101 9141 2023-05-25T11:29:34.053Z 45 no 46 Miloš, 1990- Mandić 99803657 2022 63 mentor Vladimir, 1967- Trajković 12751207 2022 63 mentor Maja, 1975- Misirkić Marjanović 99793417 2022 63 član komisije Vladimir, 1975- Perović 12897383 2022 63 član komisije Darko, 1982- Ćirić 21716583 2022 63 član komisije Biljana, 1984- Ristić 99798793 2022 63 član komisije Ljubica, 1978- Vučićević 58369289 2022 63 član komisije Silvana, 1968- Andrić 12976743 2022 86 str. 9623984 http://phaidrabg.bg.ac.rs/o:29790 no yes 12 2023-05-25T11:29:34.320Z 70 11 1066950 1066990 kutna mijeloidna leukemija, forbol-12-miristat-13-acetat, diferencijacija, autofagija,beklin-1, ERK, JNK acute myeloid leukemia, phorbol 12-myristate 13-acetate, differentiation, autophagy,beclin-1, ERK, JNK 616.155:577.2(043.3) 1738 11A32 2022