While p53-dependent apoptosis is triggered by combination of methyltransferase inhibitor decitabine (DAC) and histone deacetylase inhibitor suberoylanilide hydroxamic acid (SAHA) in leukemic cell line CML-T1, reactive oxygen species (ROS) generation as well as survivin and Bcl-2 deregulation participated in DAC + SAHA-induced apoptosis in p53-deficient HL-60 cell line. Moreover, decrease of survivin expression level is accompanied by its delocalization from centromere-related position in mitotic cells suggesting that both antiapoptotic and cell cycle regulation roles of survivin are affected by DAC + SAHA action. Addition of subtoxic concentration of all-trans-retinoic acid (ATRA) increases the efficiency of DAC + SAHA combination on viability, apoptosis induction, and ROS generation in HL-60 cells but has no effect in CML-T1 cell line. Peripheral blood lymphocytes from healthy donors showed no damage induced by DAC + SAHA + ATRA combination. Therefore, combination of ATRA with DAC and SAHA represents promising tool for therapy of leukemic disease with nonfunctional p53 signalization.

• MeSH
• apoptóza účinky léků   MeSH
• azacytidin analogy a deriváty   toxicita   MeSH
• decitabin MeSH
• down regulace účinky léků   MeSH
• HL-60 buňky MeSH
• inhibitory apoptózy metabolismus   MeSH
• kontrolní body fáze G2 buněčného cyklu účinky léků   MeSH
• kontrolní body M fáze buněčného cyklu účinky léků   MeSH
• kyseliny hydroxamové toxicita   MeSH
• lidé MeSH
• lymfocyty cytologie   účinky léků   imunologie   MeSH
• nádorové buněčné linie MeSH
• nádorový supresorový protein p53 nedostatek   genetika   MeSH
• protinádorové antimetabolity farmakologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• survivin MeSH
• synergismus léků MeSH
• tretinoin farmakologie   MeSH
• vorinostat MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• azacytidin MeSH
• BIRC5 protein, human MeSH Prohlížeč
• decitabin MeSH
• inhibitory apoptózy MeSH
• kyseliny hydroxamové MeSH
• nádorový supresorový protein p53 MeSH
• protinádorové antimetabolity MeSH
• reaktivní formy kyslíku MeSH
• survivin MeSH
• tretinoin MeSH
• vorinostat MeSH

Epigenetic therapy reverting aberrant acetylation or methylation offers the possibility to target preferentially tumor cells and to preserve normal cells. Combination epigenetic therapy may further improve the effect of individual drugs. We investigated combined action of demethylating agent decitabine and histone deacetylase inhibitor SAHA (Vorinostat) on different leukemic cell lines in comparison with peripheral blood lymphocytes. Large decrease of viability, as well as huge p21WAF1 induction, reactive oxygen species formation, and apoptotic features due to combined decitabine and SAHA action were detected in leukemic cell lines irrespective of their p53 status, while essentially no effect was observed in response to the combined drug action in normal peripheral blood lymphocytes of healthy donors. p53-dependent apoptotic pathway was demonstrated to participate in the wtp53 CML-T1 leukemic cell line response, while significant influence of reactive oxygen species on viability decrease has been detected in p53-null HL-60 cell line.

• MeSH
• apoptóza účinky léků   MeSH
• azacytidin aplikace a dávkování   analogy a deriváty   MeSH
• decitabin MeSH
• kultivované buňky MeSH
• kyseliny hydroxamové aplikace a dávkování   MeSH
• leukemie farmakoterapie   patologie   patofyziologie   MeSH
• lidé MeSH
• lymfocyty cytologie   účinky léků   fyziologie   MeSH
• protinádorové antimetabolity aplikace a dávkování   MeSH
• protokoly protinádorové kombinované chemoterapie aplikace a dávkování   MeSH
• vorinostat MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• azacytidin MeSH
• decitabin MeSH
• kyseliny hydroxamové MeSH
• protinádorové antimetabolity MeSH
• vorinostat MeSH

Reactive oxygen species play an important role in the process of apoptosis in many cell types. In this paper, we analyzed the role of ROS in DNA-damaging agents (actinomycin D or decitabine), which induced apoptosis of leukemia cell line CML-T1 and normal peripheral blood lymphocytes (PBL). The possibility of synergism with histone deacetylase inhibitors butyrate or SAHA is also reported. We found that in cancer cell line, ROS production significantly contributed to apoptosis triggering, while in normal lymphocytes treated by cytostatic or cytotoxic drugs, necrosis as well as apoptosis occurred and large heterogeneity of ROS production was measured. Combined treatment with histone deacetylase inhibitor did not potentiate actinomycin D action, whereas combination of decitabine and SAHA brought synergistic ROS generation and apoptotic features in CML cell line. Appropriate decrease of cell viability indicated promising therapeutic potential of this combination in CML, but side effects on normal PBL should be taken into attention.

• MeSH
• apoptóza účinky léků   MeSH
• azacytidin analogy a deriváty   farmakologie   MeSH
• butyráty farmakologie   MeSH
• cytostatické látky farmakologie   MeSH
• daktinomycin farmakologie   MeSH
• decitabin MeSH
• inhibitory histondeacetylas farmakologie   MeSH
• kaspasa 3 metabolismus   MeSH
• kyseliny hydroxamové farmakologie   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• poly(ADP-ribosa)polymerasy metabolismus   MeSH
• poškození DNA účinky léků   MeSH
• protinádorové látky farmakologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• synergismus léků MeSH
• vorinostat MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• azacytidin MeSH
• butyráty MeSH
• cytostatické látky MeSH
• daktinomycin MeSH
• decitabin MeSH
• inhibitory histondeacetylas MeSH
• kaspasa 3 MeSH
• kyseliny hydroxamové MeSH
• poly(ADP-ribosa)polymerasy MeSH
• protinádorové látky MeSH
• reaktivní formy kyslíku MeSH
• vorinostat MeSH

Restoration of cellular apoptotic pathways plays a crucial role in cancer therapy strategies. In a broad spectrum of anticancer drugs, epigenetic effectors are in the center of interest mostly because of potential reversibility of their action. Methylation status of the cells is influenced by methyltransferase inhibitor 2-deoxy-5'-azacytidine (decitabine, DAC), but higher concentrations of this agent cause a DNA-damage. In our study, tumor supressor p53-apoptotic pathway was activated in decitabine-induced cell death. Expression of p53-inducible BH3-only apoptotic proteins Puma and Noxa was elevated and large activation of executive caspases was observed. The extent of acetylation in the cell is affected by histonedeacetylase inhibitor suberoylanilide hydroxamic acid (SAHA). Combination of SAHA with decitabine brought synergistic effect on apoptosis triggering in CML-T1 cell line, but apoptosis as well as necrosis occurred also in normal peripheral blood lymphocytes. Therefore, promising potential of such combined therapy calls for more detailed investigation of unwanted effects in normal cells.

• MeSH
• apoptóza účinky léků   genetika   MeSH
• azacytidin analogy a deriváty   farmakologie   MeSH
• chronická myeloidní leukemie genetika   metabolismus   patologie   MeSH
• decitabin MeSH
• kultivované buňky MeSH
• kyseliny hydroxamové farmakologie   MeSH
• lidé MeSH
• lymfocyty účinky léků   metabolismus   MeSH
• preklinické hodnocení léčiv MeSH
• proteiny regulující apoptózu genetika   metabolismus   MeSH
• protinádorové antimetabolity farmakologie   MeSH
• protokoly protinádorové kombinované chemoterapie farmakologie   MeSH
• protoonkogenní proteiny c-bcl-2 genetika   metabolismus   MeSH
• protoonkogenní proteiny genetika   metabolismus   MeSH
• regulace genové exprese u leukemie účinky léků   MeSH
• synergismus léků MeSH
• upregulace účinky léků   genetika   MeSH
• vorinostat MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• azacytidin MeSH
• BBC3 protein, human MeSH Prohlížeč
• decitabin MeSH
• kyseliny hydroxamové MeSH
• PMAIP1 protein, human MeSH Prohlížeč
• proteiny regulující apoptózu MeSH
• protinádorové antimetabolity MeSH
• protoonkogenní proteiny c-bcl-2 MeSH
• protoonkogenní proteiny MeSH
• vorinostat MeSH