Caspase-12 is a molecule whose functions are still not well understood. Although its expression has been found in various tissues, specific roles have been described in only a few cases. These include the effect of caspase-12 on murine bone cell differentiation during craniofacial development. This work focused on the development of the limbs taking place through endochondral ossification, which precedes the formation of the cartilaginous growth plate. Caspase-12 was described here for the first time in growth plate chondrocytes during physiological development. Using pharmacological inhibition, caspase-12 was found to affect chondrogenesis. Limb-derived micromass cultures showed a significantly increased area of chondrogenic nodules after caspase-12 inhibition and there were changes in gene expression, the most significant of which was the reduction of Mmp9. These data point to potential new functions of caspase-12 in chondrogenesis.

• MeSH
• buněčná diferenciace MeSH
• chondrocyty * MeSH
• chondrogeneze * fyziologie   MeSH
• inhibitory kaspas farmakologie   MeSH
• kaspasa 12 * metabolismus   genetika   MeSH
• kultivované buňky MeSH
• matrixová metaloproteinasa 9 metabolismus   genetika   MeSH
• myši MeSH
• růstová ploténka růst a vývoj   MeSH
• vývojová regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Hair follicles undergo repetitive stages of cell proliferation and programmed cell death. The catagen stage of physiological apoptosis is connected with dynamic changes in morphology and alterations in gene expression. However, hair follicle apoptosis must be in balance with events in surrounding tissues, such as keratinocyte cornification, to maintain complex skin homeostasis. Several pro- and anti-apoptotic molecules in the skin have been reported but mainly in pathological states. In this investigation, apoptosis-related gene expression was examined during the first catagen stage of mouse hair follicle development by PCR arrays under physiological conditions. Postnatal stages P15 and P17, representing early and late catagen stages, were evaluated relatively to stage P6, representing the hair follicle growing phase, to demonstrate dynamics of gene activation during the catagen. Several statistically significant alterations were observed at P15 and particularly at P17. Bnip3L and caspase-12 identified by the PCR arrays at both catagen stages were additionally localized using immunofluorescence and were reported in physiological hair development for the first time.

• MeSH
• apoptóza fyziologie   MeSH
• kaspasa 12 biosyntéza   MeSH
• kůže cytologie   metabolismus   MeSH
• membránové proteiny biosyntéza   MeSH
• mitochondriální proteiny biosyntéza   MeSH
• myši MeSH
• regulace genové exprese fyziologie   MeSH
• vlasový folikul cytologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Searching for new strategies for effective elimination of human prostate cancer cells, we investigated the cooperative cytotoxic action of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and two platinum-based complexes, cisplatin or LA-12, and related molecular mechanisms. We demonstrated a notable ability of cisplatin or LA-12 to enhance the sensitivity of several human prostate cancer cell lines to TRAIL-induced cell death via an engagement of mitochondrial apoptotic pathway. This was accompanied by augmented Bid cleavage, Bak activation, loss of mitochondrial membrane potential, activation of caspase-8, -10, -9, and -3, and XIAP cleavage. RNAi-mediated silencing of Bid or Bak in Bax-deficient DU 145 cells suppressed the drug combination-induced cytotoxicity, further underscoring the involvement of mitochondrial signaling. The caspase-10 was dispensable for enhancement of cisplatin/LA-12 and TRAIL combination-induced cell death and stimulation of Bid cleavage. Importantly, we newly demonstrated LA-12-mediated enhancement of TRAIL-induced cell death in cancer cells derived from human patient prostate tumor specimens. Our results provide convincing evidence that employing TRAIL combined with cisplatin/LA-12 could contribute to more effective killing of prostate cancer cells compared to the individual action of the drugs, and offer new mechanistic insights into their cooperative anticancer action.

• MeSH
• amantadin analogy a deriváty   farmakologie   MeSH
• apoptóza účinky léků   MeSH
• cisplatina farmakologie   MeSH
• kaspasa 10 metabolismus   MeSH
• lidé MeSH
• mitochondrie účinky léků   metabolismus   MeSH
• nádory prostaty metabolismus   patologie   MeSH
• organoplatinové sloučeniny farmakologie   MeSH
• protein Bid metabolismus   MeSH
• protein TRAIL metabolismus   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Caspases are evolutionary conserved proteases traditionally known as participating in apoptosis and inflammation but recently discovered also in association with other processes such as proliferation or differentiation. This investigation focuses on caspase-12, ranked among inflammatory caspases but displaying other, not yet defined functions. A screening analysis pointed to statistically significant (P < 0.001) increase in expression of caspase-12 in a decisive period of mandibular bone formation when the original mesenchymal condensation turns into vascularized bone tissue. Immunofluorescence analysis confirmed the presence of caspase-12 protein in osteoblasts. Therefore, the osteoblastic cell line MC3T3-E1 was challenged to investigate any impact of caspase-12 on the osteogenic pathways. Pharmacological inhibition of caspase-12 in MC3T3-E1 cells caused a statistically significant decrease in expression of some major osteogenic genes, including those for alkaline phosphatase, osteocalcin and Phex. This downregulation was further confirmed by an alkaline phosphatase activity assay and by a siRNA inhibition approach. Altogether, this study demonstrates caspase-12 expression and points to its unknown physiological engagement in bone cells during the course of craniofacial development.

• Publikační typ
• časopisecké články MeSH

• MeSH
• apoptóza MeSH
• arterioskleróza etiologie   patologie   MeSH
• endoplazmatické retikulum MeSH
• kaspasy farmakologie   MeSH
• signální transdukce MeSH
• techniky in vitro MeSH
• vápníkové kanály MeSH

• Publikační typ
• abstrakty MeSH

Resveratrol (RSV) is a polyphenol antioxidant that has been shown to have neuroprotective effects. We sought molecular mechanisms that emphasize the anti-inflammatory activity of RSV in traumatic brain injury (TBI) in mice associated with endoplasmic reticulum stress (ERS). After establishing three experimental groups (sham, TBI, and TBI+RSV), we explored the results of RSV after TBI on ERS and caspase-12 apoptotic pathways. The expression levels of C/EBP homologous protein (CHOP), glucose regulated protein 78kD (GRP78), caspase-3, and caspase-12 in cortical brain tissues were assessed by western blotting. The qPCR analysis was also performed on mRNA expression of tumor necrosis factor (TNF)-α and interleukin (IL)-1β in cortical brain tissue. In addition, the expression of GRP78 in microglia (ionized calcium binding adaptor molecule 1; Iba-1) and neurons (neuronal nuclei; NeuN) was identified by immunofluorescence staining. The neurological function of mice was assessed by modified neurological severity scores (mNSS). After drug treatment, the expression of CHOP, GRP78, caspase-3 and caspase-12 decreased, and qPCR results showed that TNF-α and IL-1β were down-regulated. Immunofluorescence staining showed down-regulation of Iba-1+/GRP78+ and NeuN+/GRP78+ cells after RSV treatment. The mNSS analysis confirmed improvement after RSV treatment. RSV improved apoptosis by downregulating the ERS signaling pathway and improved neurological prognosis in mice with TBI.

• MeSH
• apoptóza účinky léků   MeSH
• buněčná smrt účinky léků   MeSH
• chaperon endoplazmatického retikula BiP * MeSH
• interleukin-1beta metabolismus   genetika   MeSH
• kaspasa 12 metabolismus   genetika   MeSH
• mikroglie účinky léků   metabolismus   patologie   MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• neurony účinky léků   patologie   metabolismus   MeSH
• neuroprotektivní látky farmakologie   terapeutické užití   MeSH
• prognóza MeSH
• proteiny tepelného šoku metabolismus   genetika   MeSH
• resveratrol * farmakologie   terapeutické užití   MeSH
• stres endoplazmatického retikula * účinky léků   MeSH
• TNF-alfa metabolismus   MeSH
• transkripční faktor CHOP metabolismus   genetika   MeSH
• traumatické poranění mozku * farmakoterapie   patologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The block of hematopoietic differentiation program in acute myeloid leukemia cells can be overcome by differentiating agent like retinoic acid, but it has several side effects. A study of other differentiation signaling pathways is therefore useful to predict potential targets of anti-leukemic therapy. We demonstrated previously that the co-treatment of HL-60 cells with Tumor necrosis factor-alpha (TNF-alpha) (1 ng/mL) and inhibitor of 5-lipoxygenase MK886 (5 microm) potentiated both monocytic differentiation and apoptosis. In this study, we detected enhanced activation of three main types of mitogen-activated protein kinases (MAPKs) (p38, c-Jun amino-terminal kinase [JNK], extracellular signal-regulated kinase [ERK]), so we assessed their role in differentiation using appropriate pharmacologic inhibitors. The inhibition of pro-apoptotic MAPKs (p38 and JNK) suppressed the effect of MK886 + TNF-alpha co-treatment. On the other hand, down-regulation of pro-survival ERK pathway led to increased differentiation. Those effects were accompanied by increased activation of caspases in cells treated by MK886 + TNF-alpha. Pan-caspase inhibitor ZVAD-fmk significantly decreased both number of apoptotic and differentiated cells. The same effect was observed after inhibition of caspase 9, but not caspase 3 and 8. To conclude, we evidenced that the activation of apoptotic processes and pathways supporting apoptosis (p38 and JNK MAPKs) is required for the monocytic differentiation of HL-60 cells.

• MeSH
• aktivace enzymů účinky léků   MeSH
• akutní promyelocytární leukemie farmakoterapie   metabolismus   patologie   MeSH
• apoptóza fyziologie   účinky léků   MeSH
• buněčná diferenciace účinky léků   MeSH
• chloromethylketony aminokyselin farmakologie   MeSH
• HL-60 buňky MeSH
• indoly farmakologie   MeSH
• inhibitory cysteinových proteinas farmakologie   MeSH
• inhibitory kaspas MeSH
• inhibitory lipoxygenas farmakologie   MeSH
• kaspasy metabolismus   MeSH
• lidé MeSH
• MAP kinasový signální systém účinky léků   MeSH
• monocyty patologie   účinky léků   MeSH
• NF-kappa B antagonisté a inhibitory   MeSH
• TNF-alfa farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Recent studies have identified molecular events characteristic of immunogenic cell death (ICD), including surface exposure of calreticulin (CRT), the heat shock proteins HSP70 and HSP90, the release of high-mobility group box protein 1 (HMGB1) and the release of ATP from dying cells. We investigated the potential of high hydrostatic pressure (HHP) to induce ICD in human tumor cells. HHP induced the rapid expression of HSP70, HSP90 and CRT on the cell surface. HHP also induced the release of HMGB1 and ATP. The interaction of dendritic cells (DCs) with HHP-treated tumor cells led to a more rapid rate of DC phagocytosis, upregulation of CD83, CD86 and HLA-DR and the release of interleukin IL-6, IL-12p70 and TNF-α. DCs pulsed with tumor cells killed by HHP induced high numbers of tumor-specific T cells. DCs pulsed with HHP-treated tumor cells also induced the lowest number of regulatory T cells. In addition, we found that the key features of the endoplasmic reticulum stress-mediated apoptotic pathway, such as reactive oxygen species production, phosphorylation of the translation initiation factor eIF2α and activation of caspase-8, were activated by HHP treatment. Therefore, HHP acts as a reliable and potent inducer of ICD in human tumor cells.

• MeSH
• adenosintrifosfát sekrece   MeSH
• aktivace enzymů imunologie   MeSH
• antigeny CD86 biosyntéza   MeSH
• apoptóza imunologie   MeSH
• CD antigeny biosyntéza   MeSH
• dendritické buňky imunologie   MeSH
• eukaryotický iniciační faktor 2 metabolismus   MeSH
• fagocytóza imunologie   MeSH
• fosforylace MeSH
• HLA-DR antigeny biosyntéza   MeSH
• hydrostatický tlak MeSH
• imunoglobuliny biosyntéza   MeSH
• interleukin-12 sekrece   MeSH
• interleukin-6 sekrece   MeSH
• kalretikulin biosyntéza   imunologie   MeSH
• kaspasa 8 metabolismus   MeSH
• lidé MeSH
• membránové glykoproteiny biosyntéza   MeSH
• membránové proteiny biosyntéza   MeSH
• nádorové buněčné linie MeSH
• nádory imunologie   MeSH
• protein HMGB1 imunologie   sekrece   MeSH
• proteiny tepelného šoku HSP70 biosyntéza   imunologie   MeSH
• proteiny tepelného šoku HSP90 biosyntéza   imunologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulační T-lymfocyty imunologie   MeSH
• stres endoplazmatického retikula imunologie   MeSH
• TNF-alfa sekrece   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Caspases are proteases traditionally associated with inflammation and cell death. Recently, they have also been shown to modulate cell proliferation and differentiation. The aim of the current research was to search for osteogenic molecules affected by caspase inhibition and to specify the individual caspases critical for these effects with a focus on proapoptotic caspases: caspase-2, -3, -6, -7, -8 and -9. Along with osteocalcin (Ocn), general caspase inhibition significantly decreased the expression of the Phex gene in differentiated MC3T3-E1 cells. The inhibition of individual caspases indicated that caspase-8 is a major contributor to the modification of Ocn and Phex expression. Caspase-2 and-6 had effects on Ocn and caspase-6 had an effect on Phex. These data confirm and expand the current knowledge about the nonapoptotic roles of caspases and the effect of their pharmacological inhibition on the osteogenic potential of osteoblastic cells.

• MeSH
• buněčné linie MeSH
• inhibitory kaspas farmakologie   MeSH
• kaspasy metabolismus   MeSH
• myši MeSH
• neutrální endopeptidasa regulující fosfáty metabolismus   MeSH
• osteoblasty cytologie   metabolismus   MeSH
• osteogeneze účinky léků   MeSH
• osteokalcin metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH