Meso-tetra(4-sulfonatophenyl)porphine (TPPS4), in combination with a light dose of 14 J cm-2, has a profound negative effect on the proliferation and viability of leukemia cells HL60 (human promyelocytic leukemia) and HEL (human erythroleukemia), the viability of normal lymphocytes and the colony-forming activity of human bone marrow progenitor cells. However, normal leukocytes (monocytes, granulocytes) are, to a large extent, resistant to photodynamic treatment (PDT). Whilst DNA fragmentation suggesting apoptosis is induced in HL60 cells, accumulation in the interphase of the cell cycle (G0/G1, G2/M) is mainly operative in the TPPS4-mediated PDT of HEL cells. The "dark" effect of TPPS4 on the cell viability is below 15% up to a concentration of 40 microM.

• MeSH
• autologní transplantace metody   MeSH
• buněčné dělení účinky léků   MeSH
• buňky kostní dřeně MeSH
• fotochemie MeSH
• fotochemoterapie * MeSH
• hematopoetické kmenové buňky účinky léků   MeSH
• HL-60 buňky MeSH
• leukemie MeSH
• lidé MeSH
• lymfocyty účinky léků   MeSH
• porfyriny farmakologie   MeSH
• radiosenzibilizující látky farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• porfyriny MeSH
• radiosenzibilizující látky MeSH
• tetraphenylporphine sulfonate MeSH Prohlížeč

Using two-dimensional electrophoresis we investigated the effect of 5-aminolevulinic acid (ALA)-based photodynamic therapy (PDT; induction with 1 mM ALA for 4 h followed by blue light dose of 18 J/cm2) on the protein expression in HL60 leukemia cells. ALA-PDT resulted in extensive qualitative and quantitative changes in the protein pattern of HL60 cell lysates. Of more than 1350 protein spots recognized on the protein maps of ALA-induced cells, seven proteins were enhanced and 17 suppressed following irradiation. Three of these, calreticulin precursor, p58 microsomal protein (ERp57) and protein disulfide isomerase (p55) have been identified by matrix-assisted laser desorption and ionization-mass spectrometry and the pI/molecular weight parameters of the affected proteins were estimated by computer analysis. The findings suggest participation of endoplasmic reticulum Ca(2+)-binding chaperones and/or Ca2+ signaling in ALA-PDT mediated cytotoxicity.

• MeSH
• akutní promyelocytární leukemie farmakoterapie   metabolismus   MeSH
• endoplazmatické retikulum účinky léků   metabolismus   MeSH
• fotochemoterapie * MeSH
• HL-60 buňky MeSH
• kyselina aminolevulová terapeutické užití   MeSH
• lidé MeSH
• molekulární chaperony metabolismus   MeSH
• nádorové proteiny metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• kyselina aminolevulová MeSH
• molekulární chaperony MeSH
• nádorové proteiny MeSH

Deletions of the long arm of chromosome 5 with common overlapping segment 5q31.1 are among the most frequent cytogenetic aberrations in myelodysplastic syndromes and acute myeloid leukemias (MDS/AML). We have constructed a YAC-based physical map of the 5q31.1 critical locus and localized the transcriptional transactivator Smad5 adjacent to loci showing consistent loss of heterozygosity in these disorders. Smad5 plays a key role along the bone morphogenetic protein-4 (BMP-4) inhibitory signalling pathway inducing embryonic hematopoiesis. Smad5 homologs Smad2 and DPC4 have recently been linked to human cancer. FISH analysis of AML-M2 cell line HL60 and of four MDS/AML patients revealed consistent hemizygous loss of the Smad5 locus. In HL60 cells, a translocation event within 5q31.1 associated with loss of adjacent material leads to disruption of the critical locus with partial retention of the 5q31.1 genomic sequences on a marker chromosome. RT-PCR sequencing analysis of the HL60 Smad5 remaining allele ruled out the functional inactivation of the gene analogous to that occurring in the Smad5 homologs DPC4 and Smad2 in cases of pancreatic and colorectal cancers. Mutational analysis of Smad5 in MDS/AML cases is in progress.

• MeSH
• akutní myeloidní leukemie genetika   MeSH
• alely MeSH
• chromozomální aberace MeSH
• DNA vazebné proteiny * MeSH
• fosfoproteiny genetika   MeSH
• genetické markery MeSH
• HL-60 buňky patologie   MeSH
• hybridizace in situ fluorescenční MeSH
• kosmidy MeSH
• lidé MeSH
• lidské chromozomy, pár 5 MeSH
• mapování chromozomů MeSH
• messenger RNA genetika   MeSH
• místa se sekvenční adresou MeSH
• molekulární sekvence - údaje MeSH
• protein Smad5 MeSH
• RNA nádorová genetika   MeSH
• sekvence nukleotidů MeSH
• trans-aktivátory * MeSH
• tumor supresorové geny * MeSH
• umělé kvasinkové chromozomy MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• DNA vazebné proteiny * MeSH
• fosfoproteiny MeSH
• genetické markery MeSH
• messenger RNA MeSH
• protein Smad5 MeSH
• RNA nádorová MeSH
• SMAD5 protein, human MeSH Prohlížeč
• trans-aktivátory * MeSH

We studied the mechanism of the cytotoxic effects of 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT; induction with 1 mM ALA for 4 h followed by a blue light dose of 18 J/cm(2)) on the human promyelocytic leukemia cell line HL60 using biochemical and electron microscopy methods. The disruption of mitochondrial membrane potential, deltapsi(m), was paralleled by a decrease in ATP level, unmasking of the mitochondrial antigen 7A6, release of cytochrome c into the cytoplasm, activation of caspases 9 and 3 and cleavage of poly(ADP-ribose) polymerase (PARP). This was followed by DNA fragmentation. These data suggest that ALA-PDT activates the mitochondrial apoptotic pathway. The level of endoplasmic reticulum Ca(2+)-binding chaperones ERp57 and ERp72 and of anti-apoptotic proteins Bcl-2 and Bcl-x(L) was decreased whereas that of Ca(2+)-binding protein calmodulin and the stress protein HSP60 was elevated following ALA-PDT. Inhibition of the initiator caspase 9, execution caspase 3 and Ca(2+)-dependent protease m-calpain, did not prevent DNA fragmentation. We conclude that, in our in vitro model, ALA-based photodynamic treatment initiates several signaling processes in HL60 cells that lead to rapidly progressing apoptosis, which is followed by slow necrosis. Two apoptotic processes proceed in parallel, one representing the mitochondrial pathway, the other involving disruption of calcium homeostasis and activation of the endoplasmic reticulum stress-mediated pathway.

• MeSH
• adenosintrifosfát metabolismus   MeSH
• aminokyseliny neutrální farmakologie   MeSH
• apoptóza účinky léků   účinky záření   MeSH
• buněčné dělení účinky léků   účinky záření   MeSH
• endoplazmatické retikulum účinky léků   účinky záření   ultrastruktura   MeSH
• fotochemoterapie metody   MeSH
• fotosenzibilizující látky farmakologie   MeSH
• fyziologický stres chemicky indukované   metabolismus   patologie   MeSH
• HL-60 buňky účinky léků   fyziologie   účinky záření   ultrastruktura   MeSH
• lidé MeSH
• membránové potenciály účinky léků   účinky záření   MeSH
• mitochondrie účinky léků   fyziologie   účinky záření   MeSH
• nádorové proteiny metabolismus   MeSH
• regulace genové exprese účinky léků   účinky záření   MeSH
• viabilita buněk účinky léků   účinky záření   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 5-aminovaleric acid MeSH Prohlížeč
• adenosintrifosfát MeSH
• aminokyseliny neutrální MeSH
• fotosenzibilizující látky MeSH
• nádorové proteiny MeSH

TNF-related apoptosis-inducing ligand (TRAIL) is a proapoptotic cytokine implicated in cancer cell surveillance. A potential of TRAIL as a cancer-specific therapeutic agent has been proposed, either as a single agent or in combination with chemotherapy. Prolonged exposure of TRAIL-sensitive leukemia cell line, wild-type (WT) HL60 cells to recombinant soluble TRAIL or to cytostatic agents, cytarabine and idarubicin, resulted in the establishment of resistant subclones with distinct phenotypic features. The TRAIL resistant HL60 subclones were characterized by decreased expression of TRAIL and TNFalpha death receptors. These resistant subclones had impaired activation of caspases 8 and 10 in response to TRAIL and TNFalpha, decreased TRAIL-induced nuclear translocation of NFkappaB RelA/p65, and dysregulation of the expression of several apoptosis regulators. Among the TRAIL resistant HL60 subclones we identified two separate phenotypes that differed in the expression of CD14, osteoprotegerin, and several apoptosis regulators. Both these TRAIL resistant HL60 subclones were resistant to TNFalpha, suggesting disruption of the extrinsic apoptotic pathway, but not to cytostatic agents, cytarabine and idarubicin. The concurrently derived HL60 subclones were cytarabine and idarubicin-resistant but remained sensitive to TRAIL-induced apoptosis. We identified distinct pathways for the development of HL60 leukemia cell resistance to apoptosis induction. These findings are relevant for the design of more effective strategies for leukemia therapy.

• MeSH
• akutní promyelocytární leukemie metabolismus   patologie   MeSH
• apoptóza * MeSH
• chemorezistence * MeSH
• cytarabin farmakologie   MeSH
• HL-60 buňky MeSH
• idarubicin farmakologie   MeSH
• kaspasy účinky léků   metabolismus   MeSH
• kinasa indukující NF-kappaB MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• protein TRAIL farmakologie   MeSH
• protein-serin-threoninkinasy účinky léků   metabolismus   MeSH
• proteiny regulující apoptózu účinky léků   metabolismus   MeSH
• receptory TNF účinky léků   metabolismus   MeSH
• rekombinantní proteiny farmakologie   MeSH
• TNF-alfa farmakologie   MeSH
• TRAIL receptory účinky léků   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cytarabin MeSH
• idarubicin MeSH
• kaspasy MeSH
• protein TRAIL MeSH
• protein-serin-threoninkinasy MeSH
• proteiny regulující apoptózu MeSH
• receptory TNF MeSH
• rekombinantní proteiny MeSH
• TNF-alfa MeSH
• TRAIL receptory MeSH

Hyiodine (high molecular weight hyaluronan combined with KI3 complex) is a new non-adhesive wound dressing which significantly improves the healing process. The aim of the study was to investigate the effects of Hyoidine on functional properties of isolated human keratinocytes and leukocytes, and on those of U937 and HL60 cell lines. While KI3 complex inhibited the viability and proliferation of the cells tested, Hyiodine did not have any significant effect. The expression of CD11b, CD62L and CD69 on PMNL, monocytes and lymphocytes, as well as the oxidative burst of blood neutrophils, were not changed. On the contrary, Hyiodine inhibited the PMA-activated oxidative burst and significantly increased the production of IL-6 and TNF-alpha by lymphocytes. It was concluded that hyaluronan content of Hyiodine reduces the toxic effect of KI3 complex on cells and speeds up the wound healing process by increasing the production of inflammatory cytokines.

• MeSH
• hemostatika farmakologie   MeSH
• HL-60 buňky MeSH
• imunitní systém cytologie   účinky léků   MeSH
• jod farmakologie   MeSH
• jodidy farmakologie   MeSH
• keratinocyty účinky léků   MeSH
• kyselina hyaluronová farmakologie   MeSH
• lidé MeSH
• U937 buňky MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• hemostatika MeSH
• hyaluronan-iodine complex MeSH Prohlížeč
• jod MeSH
• jodidy MeSH
• kyselina hyaluronová MeSH
• Lugol's solution MeSH Prohlížeč

BACKGROUND: Kaurane-type diterpenoids, obtained from various natural sources, have shown many biological activities, including anti-inflammatory and antitumor effects. Caracasine, an ent-kaurane diterpenoid isolated from the flowers of Croton micans, was shown to induce apoptosis in leukaemia cell lines. OBJECTIVE: The present study aimed to ascertain the compound's mechanism of cell death induction using two leukaemia cell lines, Jurkat E6.1 (T cell) and HL-60 (promyeloblast cells). METHODS: Cell death in Jurkat and HL60 cells were evaluated by flow cytometry for apoptosis with annexin-V/PI, mitochondrial membrane potential disturbance, changes in cell cycle, CD95 expression, caspase activation, Nuclear Factor kappa B inhibition, and differentiation into a neutrophil-like cell (dHL60). RESULTS: Caracasine (10 μM) increased the G0/G1 phase in Jurkat and arrested the cell cycle in the S phase in HL60. Caracasine increased CD95 expression (p<0.01 in Jurkat and p<0.05 in HL60) and caspase-8 activation (p<0.001 in Jurkat and p<0.05 in HL60). Caspase-9 was activated in both cell lines (p<0.001) along with the decline in mitochondrial Δψm (p<0.05 in Jurkat and p<0.001 in HL60). In HL60 cells, the kaurane induced neutrophil differentiation was assessed by CD40 expression and reactive oxygen species production. In Jurkat cells, caracasine inhibited the NF-κB pathway in cells pretreated with PHA to activate the NF-κB pathway, suggesting a possible role in inflammatory diseases. CONCLUSION: Caracasine induced apoptosis through the intrinsic and extrinsic pathways in both cell lines were evaluated which could be the leading structure for new anti-leukemic and anti-inflammatory drugs.

• Klíčová slova
• Caracasine, NF-κB, apoptosis, caracasine acid, caspases, cell cycle, differentiation, kaurane-type diterpenoids,
• MeSH
• apoptóza MeSH
• diterpeny kauranové * farmakologie   chemie   MeSH
• diterpeny * farmakologie   MeSH
• HL-60 buňky MeSH
• Jurkat buňky MeSH
• leukemie * farmakoterapie   MeSH
• lidé MeSH
• NF-kappa B metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• diterpeny kauranové * MeSH
• diterpeny * MeSH
• NF-kappa B MeSH

Although hexavalent chromium has been shown to induce apoptosis in cells cultivated in vitro, there appear to be no studies focusing on the dynamics of this process. To find out about dynamic patterns of hexavalent chromium-induced apoptosis, we treated Hep2 cells with 150 micrograms/ml potassium chromate and recorded their behavior as well as appearance of some crucial organelles using different morphological and biochemical methods. We found that Hep2 cells showed the earliest observable changes at 6 hours after the treatment (blebbing, chromatin shrinkage), with the entire apoptotic process lasting up to 24 hours. While all the observed cell features clearly prove apoptosis induced by hexavalent chromium, a typical apoptotic hallmark, DNA ladder, seems not to occur in this type of cells. On the other hand, in HL60 cells, used as a control, this ladder was observable.

• MeSH
• apoptóza účinky léků   MeSH
• buněčné jádro účinky léků   MeSH
• buněčné linie MeSH
• chrom farmakologie   MeSH
• HL-60 buňky MeSH
• lidé MeSH
• viabilita buněk MeSH
• videomikroskopie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• chrom MeSH

The effect of 5-aminolaevulinic acid-based photodynamic therapy (ALA-PDT) on the viability and proliferation of leukaemia/lymphoma cells as well as normal human lymphocytes has been investigated by flow cytometry-propidium iodide assay (FC-PI), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and bromodeoxyuridine (BrdU) incorporation and on clonogenic activity of normal human bone marrow progenitor cells by clonogenic methods. ALA-PDT (1 mM 5-ALA, 4 h, 18 J cm-2) reduces the number and/or suppressed proliferation of leukaemic cells of promyelocytic (HL60), B-cell-derived (DAUDI) and T-cell-derived (JURKAT) cell lines by 2 logs and that of the HEL erythroleukaemia cells by 77%. The effect of ALA-PDT on quiescent human lymphocytes is small (85% viable cells after ALA-PDT). The proliferation of lymphocytes subjected to ALA-PDT and induced with phytohaemagglutinin (PHA) decreases by 75% as compared to the untreated control. For normal human bone marrow progenitors, 58% of colony-forming units-granulocytes-macrophages (CFU-GM) and 55% burst-forming units-erythrocytes (BFU-E) activities are preserved.

• MeSH
• buněčné dělení účinky léků   MeSH
• buňky kostní dřeně účinky léků   MeSH
• časové faktory MeSH
• fotosenzibilizující látky farmakologie   MeSH
• hematopoetické kmenové buňky účinky léků   MeSH
• HL-60 buňky MeSH
• Jurkat buňky MeSH
• kyselina aminolevulová farmakologie   MeSH
• leukemie MeSH
• lidé MeSH
• nádorové buňky kultivované MeSH
• protoporfyriny metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• fotosenzibilizující látky MeSH
• kyselina aminolevulová MeSH
• protoporfyriny MeSH
• protoporphyrin IX MeSH Prohlížeč

Caracasine acid (CA) is an ent-3,4-seco-kaurene isolated from the plant Croton micans. Decreased cancer cell lines viability was reported upon CA treatment. The present study aimed to investigate the mechanism of CA induced cytotoxicity using two human cell lines, Jurkat E6.1 (human cell T lymphoma) and HL-60 (human acute promyelocytic leukemia). Significant increases of apoptotic cell death markers upon CA treatment were observed: annexin-V positiveness, potential mitochondrial disturbances, cell cycle changes, caspase activation, and CD95 expression. These effects were not detected in normal lymphocytes. CA induced the appearance of Bax, cleaved caspase 3, and cytochrome c release in Jurkat cells, and cleaved caspase 3 and phosphorylated p53 in HL60 cells. Likewise, downregulation of anti-apoptotic proteins such as Bcl-x (Jurkat), Bcl-2, and XIAP (HL60) was observed with CA treatment. Both pathways, intrinsic and extrinsic were activated when cell lines were treated with CA. NF-κB p65 inhibition was observed in Jurkat cells and cell differentiation in HL-60 cells. CA could be a potential leader compound for the development of new drugs for leukemia treatment in humans.

• Klíčová slova
• Annexin-V, Apoptosis, Cancer, Caracasine acid, Croton, Kaurene compounds, Leukemia, NF-κB,
• MeSH
• antitumorózní látky fytogenní farmakologie   terapeutické užití   MeSH
• apoptóza účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• Croton chemie   MeSH
• diterpeny kauranové farmakologie   terapeutické užití   MeSH
• HL-60 buňky MeSH
• Jurkat buňky MeSH
• léky antitumorózní - screeningové testy MeSH
• leukemie farmakoterapie   patologie   MeSH
• lidé MeSH
• signální transdukce účinky léků   MeSH
• transkripční faktor RelA antagonisté a inhibitory   metabolismus   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antitumorózní látky fytogenní MeSH
• caracasine acid MeSH Prohlížeč
• diterpeny kauranové MeSH
• Rela protein, mouse MeSH Prohlížeč
• transkripční faktor RelA MeSH