Apoptosis Dotaz Zobrazit nápovědu
Eryptosis is a regulated cell death (RCD) of mature erythrocytes initially described as a counterpart of apoptosis for enucleated cells. However, over the recent years, a growing number of studies have emphasized certain differences between both cell death modalities. In this review paper, we underline the hallmarks of eryptosis and apoptosis and highlight resemblances and dissimilarities between both RCDs. We summarize and critically discuss differences in the impact of caspase-3, Ca2+ signaling, ROS signaling pathways, opposing roles of casein kinase 1α, protein kinase C, Janus kinase 3, cyclin-dependent kinase 4, and AMP-activated protein kinase to highlight a certain degree of divergence between apoptosis and eryptosis. This review emphasizes the crucial importance of further studies that focus on deepening our knowledge of cell death machinery and identifying novel differences between cell death of nucleated and enucleated cells. This might provide evidence that erythrocytes can be defined as viable entities capable of programmed cell destruction. Additionally, the revealed cell type-specific patterns in cell death can facilitate the development of cell death-modulating therapeutic agents.
- Klíčová slova
- Ca2+ signaling, Casein kinase 1α, Caspase-3, Regulated cell death, p38 MAPK,
- MeSH
- apoptóza * MeSH
- buněčná smrt MeSH
- eryptóza * MeSH
- erytrocyty metabolismus MeSH
- fosfatidylseriny metabolismus MeSH
- reaktivní formy kyslíku metabolismus MeSH
- signální transdukce MeSH
- vápník metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- fosfatidylseriny MeSH
- reaktivní formy kyslíku MeSH
- vápník MeSH
Apoptosis plays an important role in the myocardial loss after acute myocardial infarction and participates in the process of subsequent left ventricular remodeling and development of symptomatic heart failure. Finding a sensitive apoptotic marker that would help in prognostic stratification of patients after acute myocardial infarction and offer new therapeutic strategies is thus of a great importance. Several studies suggest that tumor necrosis factor-related apoptosis inducing ligand (TRAIL) represents a very promising marker of prognosis in patients with acute myocardial infarction. This review article provides an overview of current knowledge on the role of apoptosis in ischemic heart disease and highlights potentially beneficial apoptotic markers in clinical practice.
- Klíčová slova
- Apoptosis, Heart failure, Ischemic heart disease, Outcome,
- MeSH
- apoptóza * MeSH
- biologické markery metabolismus MeSH
- biologické modely MeSH
- ischemická choroba srdeční metabolismus patologie MeSH
- lidé MeSH
- receptory domény smrti metabolismus MeSH
- signální transdukce MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- biologické markery MeSH
- receptory domény smrti MeSH
In malignant melanoma complex reprogramming of cell death and survival pathways leads to increased chemoresistance and poor longer-term survival. Sulforaphane (SF) is a promising isothiocyanate compound occurring in cruciferous plants with reported antiproliferative and proapoptotic activity in several tumor cell lines including melanoma. In this work we investigated the effects of SF in several melanoma cell lines and fresh melanoma cultivates. We found that SF is cytotoxic and induces mitochondrial, caspase-dependent apoptosis in our study model, however with lower efficiency in fresh melanoma cultivates. Moreover, our results indicate that in melanoma cell lines and fresh melanoma cultivates SF induces multiple signaling including oxidative stress-mediated activation of DNA-damage response pathway, changes in p38 kinase activity and enhanced expression of Bax and Puma proapoptotic proteins. In addition, in SF-exposed p53-mutant melanoma cells Puma expression seem to be under p38 control and acts as a compensatory proapoptotic mechanism. Conversely, decreased apoptosis in SF-exposed melanoma cultivates might be attributed to Akt-mediated suppression of p38 as well as p53 activity. Together, our results suggest that SF inhibits growth and proliferation and induces mitochondrial apoptosis both in melanoma cell lines as well as in fresh melanoma cultivates. This proapoptotic effect might be enhanced in combination with Akt inhibitors, in particular in melanoma samples. SF is thus commendable for further preclinical testing, both as a single agent as well as in combination regimens.
- MeSH
- antikarcinogenní látky farmakologie MeSH
- apoptóza účinky léků MeSH
- isothiokyanatany farmakologie MeSH
- kaspasy metabolismus MeSH
- lidé MeSH
- melanom patologie MeSH
- mitochondrie metabolismus MeSH
- mitogenem aktivované proteinkinasy p38 metabolismus MeSH
- nádorové buněčné linie účinky léků MeSH
- nádorový supresorový protein p53 metabolismus MeSH
- nádory kůže patologie MeSH
- reaktivní formy kyslíku metabolismus MeSH
- signální transdukce MeSH
- sulfoxidy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antikarcinogenní látky MeSH
- isothiokyanatany MeSH
- kaspasy MeSH
- mitogenem aktivované proteinkinasy p38 MeSH
- nádorový supresorový protein p53 MeSH
- reaktivní formy kyslíku MeSH
- sulforaphane MeSH Prohlížeč
- sulfoxidy MeSH
The nucleolus represents a highly dynamic nuclear compartment of the interphase nucleus. It plays a key role in ribosome biogenesis. The number of nucleoli, their size, and their activity increase in exponentially growing cells; therefore these parameters reflect the proliferating activity of the cells. A variety of staining techniques have been employed to vizualize nucleolar changes in malignant cells. Staining of so-called nucleolar organizer regions (NORs), based upon a strong avidity of nucleolar proteins to bind silver ions, represents the technique most frequently used by pathologists. Nucleolar changes and pleomorphism associated with overt proliferation of tumor cells have also been documented by immunohistochemical and ultrastructural studies. Contrary to cell proliferation, cytostatics-induced changes of nucleolar phenotype in malignant cells point to a potential role of nucleolar components in the execution of active cell death. Recent studies have provided direct clues that so-called death domains and other apoptosis-related proteins are accumulated in nucleoli upon induction of active cell death. It can be concluded that the plurifunctionality of nucleoli regarding cell proliferation and apoptosis could open new vistas toward understanding dysregulation in malignant cells.
Wnt signaling enhances cell proliferation and the maintenance of hematopoietic cells. In contrast, cytotoxic ligand Apo2L/TRAIL induces the apoptosis of various transformed cells. We observed that co-culture of human pre-B leukemia cells KM3 and REH with Wnt1- or Wnt3a-producing rat embryonic fibroblasts efficiently suppressed Apo2L/TRAIL-induced apoptosis of the lymphoid cells. This suppression occurs at the early stages of the Apo2L/TRAIL apoptotic cascade and, interestingly, the activation of the Wnt pathway alone in human leukemia cells is not sufficient for their full anti-apoptotic protection. We hypothesize that a stimulus emanating specifically from Wnt1- or Wnt3a-expressing rat fibroblasts is responsible for the observed resistance to Apo2L/TRAIL. This anti-apoptotic signaling was significantly hampered by the inhibition of the MEK1/ERK1/2 or NFkappaB pathways in KM3 and REH cells. Our results imply that paracrine Wnt-related signals could be important for the survival of pre-B cell-derived malignancies.
- MeSH
- apoptóza fyziologie MeSH
- beta-katenin fyziologie MeSH
- cykloheximid farmakologie MeSH
- daktinomycin farmakologie MeSH
- kokultivační techniky MeSH
- krysa rodu Rattus MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- pre-B-buněčná leukemie MeSH
- protein TRAIL antagonisté a inhibitory MeSH
- protein Wnt1 biosyntéza MeSH
- protein Wnt3 MeSH
- protein Wnt3A MeSH
- proteiny regulující apoptózu fyziologie MeSH
- proteiny Wnt biosyntéza MeSH
- signální transdukce fyziologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu Rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- beta-katenin MeSH
- cykloheximid MeSH
- daktinomycin MeSH
- protein TRAIL MeSH
- protein Wnt1 MeSH
- protein Wnt3 MeSH
- protein Wnt3A MeSH
- proteiny regulující apoptózu MeSH
- proteiny Wnt MeSH
- Wnt1 protein, rat MeSH Prohlížeč
- WNT3A protein, human MeSH Prohlížeč
Apoptosis has been recognized as a type of programmed cell death connected with characteristic morphological and biochemical changes in cells. This programmed cell death plays an important role in the genesis of a number of physiological and pathological processes. Thus, it can be very important to detect the signs of apoptosis in a study of cellular metabolism. The present paper provides an overview of methods often being used for detecting DNA fragmentation as one of the most specific findings in apoptosis. To date, three routine assays have been developed for detecting DNA fragmentation: DNA ladder assay, TUNEL assay, and comet assay. All these methods differ in their principles for detecting DNA fragmentation. DNA ladder assay detects the characteristic "DNA ladder" pattern formed during internucleosomal cleavage of DNA. Terminal deoxynUcleotidyl transferase Nick-End Labeling (TUNEL) assay detects DNA strand breaks using terminal deoxynucleotidyl transferase catalyzing attachment of modified deoxynucleotides on the DNA strand breaks. Comet assay can be used for detecting nucleus breakdown producing single/double-strand DNA breaks. The aim of this review is to describe the present knowledge on these three methods, including optimized approaches, techniques, and limitations.
- Klíčová slova
- Apoptosis, Apoptosis assays, Comet assay, DNA fragmentation, DNA ladder, TUNEL assay,
- MeSH
- apoptóza genetika fyziologie MeSH
- biotest metody MeSH
- DNA analýza genetika metabolismus MeSH
- fragmentace DNA * MeSH
- kometový test metody MeSH
- koncové značení zlomů DNA in situ metody MeSH
- lidé MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- DNA MeSH
We explored the mechanism of human osteosarcoma MG-63 cell apoptosis induced by asta-xanthin. The MTT assay was used to detect the effect of astaxanthin on cell viability. Morphological changes associated with apoptosis were observed after DAPI staining. Early and late stages of apoptosis were detected by flow cytometry with annexin V-FITC/PI staining. Activation of caspases-8, -9 and -3 was detected by enzyme activity in vitro. Changes in the mitochondrial membrane potential were detected by MitoCapture staining. Western blot was used to detect the cleavage of PARP, which is a caspase-3 substrate, the release of cytochrome c and Smac into the cytosol, the translocation of pro-apoptotic proteins Bax and Bak, and the expression of mitochondrial pathway-related proteins. The translocation of Bax was also detected by immunofluorescence assay. Astaxanthin significantly inhibited the viability of human osteosarcoma MG-63 cells with an IC50 value of 12.36 μg/ml. The DAPI-stained cells showed characteristic apoptotic morphological changes - cell shrinkage, cell membrane blebbing, nuclear condensation, and apoptotic body formation. Cytochrome c and Smac were released from mitochondria to the cytosol. Pro-apoptotic proteins Bax and Bak were rapidly translocated to mitochondria after six hours of astaxanthin action. Caspases-9 and -3 were activated and PARP was cleaved. The expression of anti-apoptotic proteins Bcl-2, Bcl-xL and XIAP was significantly decreased. Astaxanthin induced human osteosarcoma MG-63 cell apoptosis through the mitochondria-mediated endogenous apoptosis pathway.
- Klíčová slova
- MG-63 cells, apoptosis, astaxanthin, cytochrome c, mitochondria,
- MeSH
- apoptóza MeSH
- cytochromy c * metabolismus MeSH
- lidé MeSH
- osteosarkom * farmakoterapie metabolismus MeSH
- PARP inhibitory farmakologie terapeutické užití MeSH
- protein X asociovaný s bcl-2 metabolismus MeSH
- proteiny regulující apoptózu farmakologie terapeutické užití MeSH
- protoonkogenní proteiny c-bcl-2 metabolismus MeSH
- xanthofyly MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- astaxanthine MeSH Prohlížeč
- cytochromy c * MeSH
- PARP inhibitory MeSH
- protein X asociovaný s bcl-2 MeSH
- proteiny regulující apoptózu MeSH
- protoonkogenní proteiny c-bcl-2 MeSH
- xanthofyly MeSH
TNF-related apoptosis-inducing ligand (TRAIL) is a pro-apoptotic ligand from the TNF-alpha family that is under consideration, along with agonistic anti-TRAIL receptor antibodies, as a potential anti-tumor agent. However, most primary human tumors are resistant to monotherapy with TRAIL apoptogens, and thus the potential applicability of TRAIL in anti-tumor therapy ultimately depends on its rational combination with drugs targeting these resistances. In our high-throughput screening for novel agents/drugs that could sensitize TRAIL-resistant colorectal cancer cells to TRAIL-induced apoptosis, we found homoharringtonine (HHT), a cephalotaxus alkaloid and tested anti-leukemia drug, to be a very effective, low nanomolar enhancer of TRAIL-mediated apoptosis/growth suppression of these resistant cells. Co-treatment of TRAIL-resistant RKO or HT-29 cells with HHT and TRAIL led to the effective induction of apoptosis and the complete elimination of the treated cells. HHT suppressed the expression of the anti-apoptotic proteins Mcl-1 and cFLIP and enhanced the TRAIL-triggered activation of JNK and p38 kinases. The shRNA-mediated down-regulation of cFLIP or Mcl-1 in HT-29 or RKO cells variably enhanced their TRAIL-induced apoptosis but it did not markedly sensitize them to TRAIL-mediated growth suppression. However, with the notable exception of RKO/sh cFLIP cells, the downregulation of cFLIP or Mcl-1 significantly lowered the effective concentration of HHT in HHT + TRAIL co-treatment. Combined HHT + TRAIL therapy also led to the strong suppression of HT-29 tumors implanted into immunodeficient mice. Thus, HHT represents a very efficient enhancer of TRAIL-induced apoptosis with potential application in TRAIL-based, anti-cancer combination therapy.
- MeSH
- apoptóza účinky léků MeSH
- buňky HT-29 MeSH
- chemorezistence účinky léků MeSH
- harringtoniny farmakologie MeSH
- homoharringtonin MeSH
- lidé MeSH
- myši inbrední NOD MeSH
- myši SCID MeSH
- myši MeSH
- protein TRAIL farmakologie MeSH
- proteiny regulující apoptózu metabolismus MeSH
- transplantace heterologní MeSH
- transplantace nádorů MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- harringtoniny MeSH
- homoharringtonin MeSH
- protein TRAIL MeSH
- proteiny regulující apoptózu MeSH
Apoptosis plays crucial role in the pathogenesis of toxoplasmosis, as it limits further development of the disease. The current study aimed to investigate the effects of different concentrations of soluble total antigen (STAg) of Toxoplasma gondii (Nicolle et Manceaux, 1908) on the apoptotic and anti-apoptotic pathways. PMA-activated THP-1 cell line was sensed by T. gondii STAg and the expression patterns of caspase-3, -7, -8, -9, Bax, Bcl-2, and Mcl-1 genes were evaluated. The results showed statistically significant concentration-dependent overexpression of both Bcl-2 (P-value < 0.0001) and Mcl-1 (P-value = 0.0147). The cas-7 showed overexpression in all concentrations (P-value < 0.0001). The cas-3 was suppressed in concentrations 100, 80, and 40 µg, but statistically significant downregulated in concentrations 10 and 20 µg. The Bax was suppressed in concentrations 100 to 20 µg, while it slightly downregulated 1.42 fold (P-value = 0.0029) in concentration 10 µg. The expression of cas-8 and -9 was suppressed in all concentrations. Our results indicated that T. gondii STAg downregulated and suppressed apoptotic and upregulated anti-apoptotic pathways. The upregulation of cas-7 in this study may indicate the role of T. gondii STAg in activation of inflammatory responses.
- Klíčová slova
- Apoptosis, Bcl-2., Mcl-1, Soluble total antigen, Toxoplasma gondii,
- MeSH
- apoptóza MeSH
- buněčné linie MeSH
- lidé MeSH
- monocyty MeSH
- Toxoplasma * MeSH
- toxoplazmóza * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
We studied the cellular localization of the apoptotic proteins endonuclease G, AIF, and AMID in silico using three prediction tools and in living cells using both single-cell colocalization image analysis and nuclear translocation analysis. We confirmed the mitochondrial localization of endonuclease G and AIF by prediction analysis and by single-cell colocalization image analysis. We found the AMID protein to be cytoplasmic, most probably incorporated into the cytoplasmic side of the membranes of various organelles. The highest concentration of AMID was observed associated with the Golgi. Colocalization of AMID with lysosomes was also indirectly confirmed by analysis of AMID-rich vesicle velocity using manual tracking analysis. Bioinformatic analysis also detected nuclear localization signals in endonuclease G and AIF, but not in AMID. A novel analysis of time-lapse fluorescence image data during staurosporine-induced apoptosis revealed nuclear translocation only for endonuclease G and AIF.
- MeSH
- apoptóza fyziologie MeSH
- buněčné jádro metabolismus MeSH
- buněčné linie MeSH
- endodeoxyribonukleasy izolace a purifikace metabolismus MeSH
- faktor vyvolávající apoptózu izolace a purifikace metabolismus MeSH
- fluorescenční mikroskopie MeSH
- Golgiho aparát metabolismus MeSH
- intracelulární membrány metabolismus MeSH
- lidé MeSH
- lyzozomy metabolismus MeSH
- mitochondriální proteiny izolace a purifikace metabolismus MeSH
- mitochondrie metabolismus MeSH
- proteiny regulující apoptózu izolace a purifikace metabolismus MeSH
- rekombinantní fúzní proteiny metabolismus MeSH
- výpočetní biologie * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- AIFM1 protein, human MeSH Prohlížeč
- endodeoxyribonukleasy MeSH
- endonuclease G MeSH Prohlížeč
- faktor vyvolávající apoptózu MeSH
- ferroptosis suppressor protein 1, human MeSH Prohlížeč
- mitochondriální proteiny MeSH
- proteiny regulující apoptózu MeSH
- rekombinantní fúzní proteiny MeSH
