The main objective of this study was to analyze changes in the antiproliferative effect of vitamin D3, in the form of calcitriol and calcidiol, via its combined application with all-trans retinoic acid (ATRA) in osteosarcoma cell lines. The response to treatment with calcitriol and calcidiol alone was specific for each cell line. Nevertheless, we observed an enhanced effect of combined treatment with ATRA and calcitriol in the majority of the cell lines. Although the levels of respective nuclear receptors did not correlate with the sensitivity of cells to these drugs, vitamin D receptor (VDR) upregulation induced by ATRA was found in cell lines that were the most sensitive to the combined treatment. In addition, all these cell lines showed high endogenous levels of retinoic acid receptor α (RARα). Our study confirmed that the combination of calcitriol and ATRA can achieve enhanced antiproliferative effects in human osteosarcoma cell lines in vitro. Moreover, we provide the first evidence that ATRA is able to upregulate VDR expression in human osteosarcoma cells. According to our results, the endogenous levels of RARα and VDR could be used as a predictor of possible synergy between ATRA and calcitriol in osteosarcoma cells.

• Klíčová slova
• all-trans retinoic acid, calcidiol, calcitriol, osteosarcoma, retinoic acid receptor α, vitamin D receptor,
• MeSH
• alfa receptor kyseliny retinové metabolismus   MeSH
• antitumorózní látky aplikace a dávkování   MeSH
• kalcifediol aplikace a dávkování   MeSH
• kalcitriol aplikace a dávkování   MeSH
• léky antitumorózní - screeningové testy MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• osteosarkom farmakoterapie   metabolismus   MeSH
• protokoly antitumorózní kombinované chemoterapie MeSH
• receptory kalcitriolu metabolismus   MeSH
• tretinoin aplikace a dávkování   MeSH
• vitaminy aplikace a dávkování   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alfa receptor kyseliny retinové MeSH
• antitumorózní látky MeSH
• kalcifediol MeSH
• kalcitriol MeSH
• receptory kalcitriolu MeSH
• tretinoin MeSH
• vitaminy MeSH

Retinoic acid (RA) is able to induce the differentiation of embryonic stem cells into neuronal lineages. The mechanism of this effect is unknown but it has been evidenced to be dependent on the formation of floating spheroids called embryoid bodies. Results presented here show that the inhibition of phosphoinositide 3-kinase signaling pre-determines mouse embryonic stem cells to RA induced neurogenesis in monolayer culture with no need of embryoid bodies formation.

• MeSH
• 1-fosfatidylinositol-3-kinasa metabolismus   MeSH
• buněčné kultury MeSH
• chromony farmakologie   MeSH
• diferenciační antigeny genetika   metabolismus   MeSH
• embryonální kmenové buňky účinky léků   metabolismus   fyziologie   MeSH
• genetická transkripce MeSH
• inhibitory fosfoinositid-3-kinasy * MeSH
• kadheriny genetika   metabolismus   MeSH
• keratin-8 genetika   metabolismus   MeSH
• kultivované buňky MeSH
• luciferasy biosyntéza   genetika   MeSH
• molekuly buněčné adheze nervové genetika   metabolismus   MeSH
• morfoliny farmakologie   MeSH
• myši MeSH
• neurogeneze účinky léků   MeSH
• reportérové geny MeSH
• signální transdukce účinky léků   MeSH
• tretinoin farmakologie   MeSH
• tubulin genetika   metabolismus   MeSH
• tvar buňky účinky léků   MeSH
• vývojová regulace genové exprese účinky léků   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 1-fosfatidylinositol-3-kinasa MeSH
• 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one MeSH Prohlížeč
• beta3 tubulin, mouse MeSH Prohlížeč
• chromony MeSH
• diferenciační antigeny MeSH
• inhibitory fosfoinositid-3-kinasy * MeSH
• kadheriny MeSH
• keratin-8 MeSH
• luciferasy MeSH
• molekuly buněčné adheze nervové MeSH
• morfoliny MeSH
• tretinoin MeSH
• tubulin MeSH

The interaction between retinoids and transforming growth factor-beta1 (TGF-beta1) leading to regulation of proliferation, differentiation and apoptosis is not still fully understood. In this study, we demonstrated that a combination treatment with all-trans retinoic acid (ATRA) and TGF-beta1 led to the enhancement of ATRA-induced suppression of cell proliferation, which is accompanied by inhibition of ATRA-induced apoptosis in human leukemia HL-60 cells. This effect was preceded by the arrest of cells in G0/G1 cell cycle phase linked with pRb protein dephosphorylation, continuous accumulation of p21 and transiently increased level of p27, inhibitors of cyclin-dependent kinases. Inhibition of ATRA-induced apoptosis by TGF-beta1 was associated with an increased level of Mcl-1 protein, an anti-apoptotic member of Bcl-2 family, but not with inhibition of mitochondrial membrane depolarization. Levels of other Bcl-2 family proteins (Bcl-2, Bcl-X(L), Bad, Bak, Bax) were unaffected by simultaneous ATRA and TGF-beta1 treatment, when compared to ATRA alone. Upregulation of c-FLIP(L) protein, an inhibitor of apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), correspond with inhibition of ATRA-induced (autocrine TRAIL-mediated) caspase-8 activation and apoptosis. These results suggest that apoptosis inhibition associated with proliferation block could depend on modulation of the TRAIL apoptotic pathway and regulation of the Mcl-1 protein level. In summary, we demonstrate that the balance of processes leading to regulation of proliferation and differentiation of myeloid cells can modulate cell sensitivity to apoptosis-inducing stimuli.

• MeSH
• aktivace enzymů účinky léků   MeSH
• antigeny CD11b biosyntéza   účinky léků   MeSH
• apoptóza účinky léků   fyziologie   MeSH
• buněčná diferenciace účinky léků   MeSH
• buněčný cyklus účinky léků   MeSH
• FLIP (buněčný) MeSH
• fosforylace MeSH
• G0 fáze účinky léků   MeSH
• G1 fáze účinky léků   MeSH
• granulocyty účinky léků   fyziologie   MeSH
• HL-60 buňky MeSH
• inhibitor p21 cyklin-dependentní kinasy biosyntéza   účinky léků   MeSH
• intracelulární signální peptidy a proteiny účinky léků   metabolismus   MeSH
• kaspasa 3 MeSH
• kaspasa 8 MeSH
• kaspasy účinky léků   metabolismus   MeSH
• lidé MeSH
• membránové glykoproteiny metabolismus   farmakologie   MeSH
• mitochondriální membrány účinky léků   fyziologie   MeSH
• nádorové buňky kultivované MeSH
• nádorové proteiny účinky léků   metabolismus   MeSH
• proliferace buněk účinky léků   MeSH
• protein MCL-1 MeSH
• protein TRAIL MeSH
• protein X asociovaný s bcl-2 účinky léků   metabolismus   MeSH
• proteiny regulující apoptózu metabolismus   farmakologie   MeSH
• protoonkogenní proteiny c-bcl-2 účinky léků   metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• retinoblastomový protein účinky léků   metabolismus   MeSH
• synergismus léků MeSH
• TNF-alfa metabolismus   farmakologie   MeSH
• transformující růstový faktor beta farmakologie   MeSH
• transformující růstový faktor beta1 MeSH
• tretinoin antagonisté a inhibitory   farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny CD11b MeSH
• BAX protein, human MeSH Prohlížeč
• CASP3 protein, human MeSH Prohlížeč
• CASP8 protein, human MeSH Prohlížeč
• CDKN1A protein, human MeSH Prohlížeč
• CFLAR protein, human MeSH Prohlížeč
• FLIP (buněčný) MeSH
• inhibitor p21 cyklin-dependentní kinasy MeSH
• intracelulární signální peptidy a proteiny MeSH
• kaspasa 3 MeSH
• kaspasa 8 MeSH
• kaspasy MeSH
• membránové glykoproteiny MeSH
• nádorové proteiny MeSH
• protein MCL-1 MeSH
• protein TRAIL MeSH
• protein X asociovaný s bcl-2 MeSH
• proteiny regulující apoptózu MeSH
• protoonkogenní proteiny c-bcl-2 MeSH
• reaktivní formy kyslíku MeSH
• retinoblastomový protein MeSH
• TGFB1 protein, human MeSH Prohlížeč
• TNF-alfa MeSH
• TNFSF10 protein, human MeSH Prohlížeč
• transformující růstový faktor beta MeSH
• transformující růstový faktor beta1 MeSH
• tretinoin MeSH

Alterations of cell surface expression of HLA (class I, class II DR, DP and DQ) and EGF-receptor on two malignant glioma cell lines (U-343MG and U-563MG) induced with cytokines (IFN-gamma, TNF-alpha, IL-1 alpha) and differentiation promoters (all-trans retinoic acid, phorbol ester TPA) were analyzed with the aid of flow cytometry. IFN-gamma induced a 10-15fold increase of HLA class I. TNF-alpha alone induced a two- to fivefold increase of HLA class I cell surface density and increased the IFN-gamma induced upregulation of HLA class I to approximately 20-24 times the antigen density of uninduced cells. TNF-alpha was able to increase HLA class II DR and DP cell surface expression on glioma lines, but it enhanced only the IFN-gamma-induced HLA class II DR upregulation. All-trans retinoic acid and TPA regulated in the opposite way the EGF-receptor cell surface expression on U-563MG cells.

• MeSH
• antigeny povrchové účinky léků   metabolismus   MeSH
• buněčná diferenciace účinky léků   MeSH
• cytokiny metabolismus   farmakologie   MeSH
• erbB receptory účinky léků   fyziologie   MeSH
• gliom chemie   imunologie   MeSH
• HLA antigeny účinky léků   fyziologie   MeSH
• interferon gama metabolismus   farmakologie   MeSH
• interleukin-1 metabolismus   farmakologie   MeSH
• lidé MeSH
• nádorové buňky kultivované účinky léků   MeSH
• tetradekanoylforbolacetát farmakologie   MeSH
• TNF-alfa metabolismus   farmakologie   MeSH
• tretinoin farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny povrchové MeSH
• cytokiny MeSH
• erbB receptory MeSH
• HLA antigeny MeSH
• interferon gama MeSH
• interleukin-1 MeSH
• tetradekanoylforbolacetát MeSH
• TNF-alfa MeSH
• tretinoin MeSH

Differentiating myeloid cells may become resistant to various apoptotic stimuli. In the present study, dimethyl sulfoxide (DMSO) and all-trans retinoic acid (ATRA) were found to modulate the sensitivity of HL-60 cells to death receptor-mediated apoptosis in a time-dependent manner. During the early stages of differentiation, DMSO treatment increased the response of HL-60 cells to tumor necrosis factor alpha; (TNF-alpha), but enhanced responsiveness was lost during later differentiation stages. In contrast, ATRA treatment induced resistance to TNF-alpha-induced apoptosis. HL-60 cells were resistant to Fas-mediated apoptosis but were sensitized by culturing in serum-free conditions. Similar to its effect on TNF-alpha sensitivity, DMSO pretreatment augmented the response to Fas-mediated signaling, which coincided with increased expression of Fas on DMSO-pretreated cells. However, during the later stages of DMSO-induced differentiation, sensitivity to anti-Fas antibody-induced apoptosis declined significantly, although Fas expression was still elevated. The reduced sensitivity to anti-Fas treatment partially correlated with increased Fas-associated phosphatase-1 mRNA expression. Thus, regardless of either Fas up-regulation or potentiation of TNF-alpha-mediated apoptosis during early DMSO-induced differentiation, a slow increase in resistance to apoptosis mediated through these death receptors occurs during DMSO-induced differentiation, which contrasts with the rapid induction of resistance following treatment with ATRA.

• MeSH
• antigeny CD95 metabolismus   MeSH
• apoptóza * MeSH
• buněčná diferenciace MeSH
• CD antigeny biosyntéza   MeSH
• dimethylsulfoxid farmakologie   MeSH
• HL-60 buňky MeSH
• kinetika MeSH
• kultivační média bez séra MeSH
• lidé MeSH
• messenger RNA MeSH
• monoklonální protilátky metabolismus   MeSH
• myeloidní leukemie MeSH
• proteinfosfatasa 1 MeSH
• protoonkogenní proteiny c-bcl-2 biosyntéza   metabolismus   MeSH
• receptory TNF - typ II MeSH
• receptory TNF biosyntéza   MeSH
• TNF-alfa farmakologie   MeSH
• transportní proteiny genetika   metabolismus   MeSH
• tretinoin farmakologie   MeSH
• tyrosinfosfatasa nereceptorového typu 13 MeSH
• tyrosinfosfatasy genetika   metabolismus   MeSH
• upregulace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny CD95 MeSH
• CD antigeny MeSH
• dimethylsulfoxid MeSH
• kultivační média bez séra MeSH
• messenger RNA MeSH
• monoklonální protilátky MeSH
• proteinfosfatasa 1 MeSH
• protoonkogenní proteiny c-bcl-2 MeSH
• PTPN13 protein, human MeSH Prohlížeč
• receptory TNF - typ II MeSH
• receptory TNF MeSH
• TNF-alfa MeSH
• transportní proteiny MeSH
• tretinoin MeSH
• tyrosinfosfatasa nereceptorového typu 13 MeSH
• tyrosinfosfatasy MeSH

We investigated whether and how could various modulators of arachidonic acid metabolism affect apoptosis induced by tumour necrosis factor-alpha (TNF-alpha) in human myeloid leukaemia HL-60 cells. These included arachinonyltrifluoromethyl ketone (AACOCF3; cytosolic phospholipase A2 inhibitor), indomethacin (cyclooxygenase inhibitor), MK-886 (3-[1-(4-chlorobenzyl)-3-t-butyl-thio-5-isopropylindol-2-yl]-2,2-dimethyl propanoic acid; 5-lipoxygenase-activating protein inhibitor), nordihydroguaiaretic acid (general lipoxygenase inhibitor), and arachidonic acid itself. Incubation of HL-60 cells with nordihydroguaiaretic acid resulted in apoptosis and it was characterised by mitochondria membrane depolarisation, release of cytochrome c from mitochondria into cytosol and activation of caspase-3. Indomethacin and nordihydroguaiaretic acid synergistically potentiated TNF-alpha-induced apoptosis, while arachidonic acid, AACOCF3 and MK-886 did not modulate its effects. Furthermore, indomethacin potentiated apoptosis in cells treated with a differentiating agent, all-trans retinoic acid, which induces resistance to TNF-alpha. However, the observed effects were probably not associated either with the cyclooxygenase- or lipoxygenase-dependent activities of indomethacin and nordihydroguaiaretic acid, respectively. Since indomethacin may reportedly activate peroxisome proliferator-activated receptors (PPARs), the effects of specific ligands of PPARs on apoptosis were studied as well. It was found that selective PPARs ligands had no effects on TNF-alpha-induced apoptosis. The findings suggest that arachidonic acid metabolism does not play a key role in regulation of apoptosis induced by TNF-alpha in the present model. Nevertheless, our data raise the possibility that indomethacin could potentially be used to improve the treatment of human myeloid leukaemia.

• MeSH
• apoptóza účinky léků   MeSH
• cyklooxygenasa 2 MeSH
• cyklooxygenasy účinky léků   metabolismus   MeSH
• cytochromy skupiny c účinky léků   metabolismus   MeSH
• cytosol účinky léků   enzymologie   MeSH
• fosfolipasy A účinky léků   metabolismus   MeSH
• fosfolipasy A2 MeSH
• HL-60 buňky MeSH
• indomethacin farmakologie   MeSH
• inhibitory cyklooxygenasy 2 MeSH
• inhibitory cyklooxygenasy farmakologie   MeSH
• izoenzymy účinky léků   metabolismus   MeSH
• kaspasa 3 MeSH
• kaspasy účinky léků   metabolismus   MeSH
• kyselina arachidonová antagonisté a inhibitory   metabolismus   MeSH
• kyselina nordihydroguaiaretová farmakologie   MeSH
• lidé MeSH
• membránové proteiny MeSH
• proliferátory peroxizomů farmakologie   MeSH
• pyrimidiny farmakologie   MeSH
• receptory cytoplazmatické a nukleární účinky léků   metabolismus   MeSH
• synergismus léků MeSH
• thiazolidindiony * MeSH
• thiazoly farmakologie   MeSH
• TNF-alfa farmakologie   MeSH
• transkripční faktory účinky léků   metabolismus   MeSH
• tretinoin farmakologie   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CASP3 protein, human MeSH Prohlížeč
• ciglitazone MeSH Prohlížeč
• cyklooxygenasa 2 MeSH
• cyklooxygenasy MeSH
• cytochromy skupiny c MeSH
• fosfolipasy A MeSH
• fosfolipasy A2 MeSH
• indomethacin MeSH
• inhibitory cyklooxygenasy 2 MeSH
• inhibitory cyklooxygenasy MeSH
• izoenzymy MeSH
• kaspasa 3 MeSH
• kaspasy MeSH
• kyselina arachidonová MeSH
• kyselina nordihydroguaiaretová MeSH
• membránové proteiny MeSH
• pirinixic acid MeSH Prohlížeč
• proliferátory peroxizomů MeSH
• PTGS2 protein, human MeSH Prohlížeč
• pyrimidiny MeSH
• receptory cytoplazmatické a nukleární MeSH
• thiazolidindiony * MeSH
• thiazoly MeSH
• TNF-alfa MeSH
• transkripční faktory MeSH
• tretinoin MeSH