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1.
Treigyté G Navakauskiené R Kulyté A Gineitis A Magnusson KE 《Cellular and molecular life sciences : CMLS》2000,57(13-14):1997-2008
Two-dimensional electrophoretic analysis was used to assess quantitative and qualitative changes in the expression and tyrosine phosphorylation of cytoplasmic proteins of proliferating, differentiating HL-60 cells and mature human blood neutrophils. The total tyrosine phosphorylation level of cytoplasmic proteins appeared approximately constant during the pre-commitment period, i.e., 6-24 h after induction of differentiation by 700 nM all-trans retinoic acid. At the time of granulocytic phenotype formation (48-120 h), the total level of tyrosine phosphorylation of cytoplasmic proteins increased significantly. Tyrosine phosphorylation of cytoplasmic proteins in matured blood neutrophils was significantly lower than that of cytoplasmic proteins of HL-60 cells differentiated for 96 h with retinoic acid. Immunoblotting with anti-Erk2 and anti-phosphotyrosine monoclonal IgG2bk antibodies showed that Erk2 was expressed and tyrosine-phosphorylated at different levels in HL-60 proliferating cells and in cells at all stages of differentiation. Our data showed that tyrosine phosphorylation of cytoplasmic proteins in differentiating HL-60 cells changes dramatically during the period of phenotype formation and is accompanied by increasing activity of Erk2. An increasing number of apoptotic cells appeared in the differentiating HL-60 cell population during the granulocyte maturation stage (48-120 h of differentiation). The appearance at this time of differentiation of a new set of tyrosine-phosphorylated cytoplasmic proteins (also distinctive for apoptotic HL-60 cells mediated by etoposide) together with an increasing number of apoptotic cells in the differentiating population strongly suggests that these proteins are associated with the apoptotic process. 相似文献
2.
Protein kinases mediate nitric oxide-induced apoptosis in the insect cell line IPLB-LdFB 总被引:2,自引:0,他引:2
The involvement of protein kinases (PKA, PKC and PKB) in nitric oxide (NO)-induced apoptosis with sodium nitroprusside plus
N-acetyl-L-cysteine in the IPLB-LdFB cell line from the insect Lymantria dispar was investigated. The presence of protein kinase-like molecules was demonstrated by western blot analysis. The role of the
kinases in programmed cell death was analysed in cytofluorimetric experiments by incubating the insect cells with H-89 (a
specific inhibitor of PKA), calphostin C (an inhibitor of PKC) or wortmannin (an inhibitor of phosphatidylinositol 3-kinase).
The results show that PKA is correlated with the induction and PKC and PKB with the prevention of NO-induced insect cell death.
Moreover, NO-induced apoptosis involves the release of cytochrome c.
Received 15 March 2002; accepted 25 March 2002 相似文献
3.
Mechanisms of p53-mediated apoptosis 总被引:25,自引:0,他引:25
4.
Anandamide induces cell death independently of cannabinoid receptors or vanilloid receptor 1: possible involvement of lipid rafts 总被引:6,自引:0,他引:6
Anandamide triggers various cellular activities by binding to cannabinoid (CB1/CB2) receptors or vanilloid receptor 1 (VR1). However, the role of these receptors in anandamide-induced apoptosis remains largely unknown. Here, we show that SR141716A, a specific inhibitor of cannabinoid receptor (CB1-R), did not block anandamide-induced cell death in endogenously CB1-R expressing cells. In addition, CB1-R-lacking Chinese hamster ovary (CHO) cells underwent cell death after anandamide treatment. SR144528, a specific inhibitor of CB2-R also failed to block anandamide-induced cell death in HL-60 cells. Capsazepine, a specific antagonist of VR1 could not prevent anandamide-induced cell death in constitutively and endogenously VR1 expressing PC12 cells. Moreover, anandamide noticeably triggered cell death in VR1-lacking human embryonic kidney (HEK) cells. In contrast, methyl-beta cyclodextrin (MCD), a membrane cholesterol depletor, completely blocked anandamide-induced cell death in a variety of cells, including PC12, C6, Neuro-2a, CHO, HEK, SMC, Jurkat and HL-60 cells. MCD also blocked anandamide-induced superoxide generation, phosphatidyl serine exposure and p38 MAPK/JNK activation. Thus, our data imply a novel role for of membrane lipid rafts in anandamide-induced cell death. 相似文献
5.
Gineitis A Treigyte G Savickiene J Shanbhag VP Stigbrand T 《Cellular and molecular life sciences : CMLS》1999,55(2):317-326
The two-dimensional electrophoretic patterns of nuclear proteins and their tyrosine phosphorylation were compared for HL-60
cells before and after differentiation induction to granulocytes by dimethyl sulfoxide, all-trans retinoic acid and N
6,O
2-dibutyryl adenosine 3′5′-cyclic monophosphate. Regardless of the inducer used, some nuclear proteins, which are tyrosine-phosphorylated
in proliferating HL-60 cells, undergo gradual dephosphorylation 12–72 h after induction of differentiation, followed by drastic
dephosphorylation during maturation to granulocytes. At least 13 nuclear proteins with a molecular mass of 35–110 kDa are
dephosphorylated, and 6 nuclear proteins undergo tyrosine phosphorylation. Analysis of the nuclear proteins differentially
extracted by salt and detergents indicates that changes in their tyrosine phosphorylation during the maturation stage of differentiating
granulocytes occur mainly in proteins which are abundant in nucleoplasm, chromatin and residual nuclear structures. The abundance
of these proteins, residing in the nuclear structures, and their long-term modification in phosphorylation during the maturation
stages of differentiation strongly suggest that tyrosine phosphorylation of these proteins is involved in reorganization of
the differentiating cell nucleus.
Received 21 September 1998; received after revision 24 November 1998; accepted 3 December 1998 相似文献
6.
The human leukemia cell line, THP-1: a multifacetted model for the study of monocyte-macrophage differentiation 总被引:8,自引:0,他引:8
J Auwerx 《Experientia》1991,47(1):22-31
THP-1 is a human monocytic leukemia cell line. After treatment with phorbol esters, THP-1 cells differentiate into macrophage-like cells which mimic native monocyte-derived macrophages in several respects. Compared to other human myeloid cell lines, such as HL-60, U937, KG-1, or HEL cell lines, differentiated THP-1 cells behave more like native monocyte-derived macrophages. Because of these characteristics, the THP-1 cell line provides a valuable model for studying the mechanisms involved in macrophage differentiation, and for exploring the regulation of macrophage-specific genes as they relate to physiological functions displayed by these cells. 相似文献
7.
J. Auwerx 《Cellular and molecular life sciences : CMLS》1991,47(1):22-31
Summary THP-1 is a human monocytic leukemia cell line. After treatment with phorbol esters, THP-1 cells differentiate into macrophage-like cells which mimic native monocyte-derived macrophages in several respects. Compared to other human myeloid cell lines, such as HL-60, U937, KG-1, or HEL cell lines, differentiated THP-1 cells behave more like native monocyte-derived macrophages. Because of these characteristics, the THP-1 cell line provides a valuable model for studying the mechanisms involved in macrophage differentiation, and for exploring the regulation of macrophage-specific genes as they relate to physiological functions displayed by these cells. 相似文献
8.
T cell activation is enhanced by the costimulatory interaction of B7 on antigen-presenting cells and CD28 on T cells, resulting
in long-term T cell proliferation, differentiation and production of large amounts of cytokines, such as interleukin (IL)-2.
CTLA-4 is a co-stimulation receptor that shares 31% homology with CD28 and binds B7 family members with higher affinity. CTLA-4
is transiently expressed intracellularly and on the cell surface following activation of T cells. We have studied the kinetics
of CTLA-4 expression and the effects of dexamethasone on CTLA-4 expression during T cell activation in cultures of mouse spleen
cells stimulated by a mixture of immobilized anti-CD3 and anti-CD28 monoclonal antibodies (anti-CD3/CD28 mAb) or concanavalin
A (ConA). CTLA-4 expression peaked on day 2 and returned to background levels after 7 days. Dexamethasone was found to potentiate
CTLA-4 expression in a dose-dependent manner with an EC50 effective concentration 50%) of about 10−8 M. In contrast, other immunosuppressive agents, such as rapamycin or cyclosporin A had no or an inhibitory effect on CTLA-4
expression, respectively. Dexamethasone also stimulated CD28 expression, but inhibited IL-2R expression during anti-CD3/CD28
mAb-induced mouse splenic T cell activation. Western blot analyses of lysates of activated mouse T cells showed that dexamethasone
increased CTLA-4 protein levels twofold during anti-CD3/CD28 mAb-induced activation. Dexamethasone also enhanced CTLA-4 messenger
RNA twofold as quantified by ribonuclease protection assay. The effects of dexamethasone on CTLA-4 expression were glucocorticoid-specific
and completely inhibited by the glucocorticoid receptor antagonist mifepristone (RU486), indicating that the effect of dexamethasone
on CTLA-4 expression is mediated through the glucocorticoid receptor. In conclusion, the immunosuppressive agent dexamethasone
actually stimulates CTLA-4 expression, which is involved in downregulation of T cell activation.
Received 19 May 1999; received after revision 13 July 1999; accepted 13 July 1999 相似文献
9.
Goh YC Yap CT Huang BH Cronshaw AD Leung BP Lai PB Hart SP Dransfield I Ross JA 《Cellular and molecular life sciences : CMLS》2011,68(9):1581-1592
Heat-shock protein 60 (Hsp60) is a highly conserved stress protein which has chaperone functions in prokaryotes and mammalian
cells. Hsp60 is associated with the mitochondria and the plasma membrane through phosphorylation by protein kinase A, and
is incorporated into lipid membranes as a protein-folding chaperone. Its diverse intracellular chaperone functions include
the secretion of proteins where it maintains the conformation of precursors and facilitates their translocation through the
plasma membrane. We report here that Hsp60 is concentrated in apoptotic membrane blebs and translocates to the surface of
cells undergoing apoptosis. Hsp60 is also enriched in platelets derived from terminally differentiated megakaryocytes and
expressed at the surface of senescent platelets. Furthermore, the exposure of monocytic U937 cells to Hsp60 enhanced their
phagocytic activity. Our results suggests that externalized Hsp60 in apoptotic cells and senescent platelets influences events
subsequent to apoptosis, such as the clearance of apoptotic cells by phagocytes. 相似文献
10.
Bártová E Jirsová P Fojtová M Soucek K Kozubek S 《Cellular and molecular life sciences : CMLS》2003,60(5):979-990
The nuclear architecture of selected chromosomes in apoptotic nuclei of human leukemic cells K-562 and HL-60 was investigated. Etoposide and prolonged confluence were used for the induction of apoptosis. DAPI as well as TUNEL labeling of apoptotic nuclear bodies was combined with visualization of chromosomal territories by the FISH technique. Simultaneous vital staining by annexin V, propidium iodide, and Hoechst 33342 was applied to distinguish apoptotic, necrotic, and intact cell fraction of tested populations. Our FISH analyses revealed that the three-dimensional (3D) structure of apoptotic nuclei as well as the 3D structure of apoptotic bodies is preserved in formaldehyde-fixed cells. High-molecular-weight DNA fragmentation was determined in apoptotic K-562 cells in contrast to oligonucleosomal cleavage observed in apoptotic HL-60 cells. In K-562 populations, chromosomal territories were located separately either in one apoptotic body or underwent disassembly into chromosomal segments dispersed into single and/or several apoptotic bodies. The apoptotic disorganization of chromosomal territories was irregular, leading mainly to chromosomal segments of different sizes and, consequently, chromosomal disassembly was not observed at specific sites. In comparison with the control, an increased number of centromeric FISH signals were observed in prolonged confluence-treated K-562 cells induced to apoptosis. This finding can be explained either as a consequence of apoptosis or by polyploidization. Sequential staining of the same apoptotic nuclei by the FISH and TUNEL techniques revealed that chromosomal territory segmentation precedes the formation of nuclear apoptotic bodies. 相似文献
11.
Corti S Salani S Del Bo R Torrente Y Strazzer S Belicchi M Paganoni S Li Z Comi GP Bresolin N Paulin D Scarlato G 《Cellular and molecular life sciences : CMLS》2001,58(1):135-140
The generation of human myogenic cell lines could potentially provide a valuable source for cell transplantation in myopathies. The dysregulation of proliferative-differentiative signals by viral oncogenes can result in the induction of apoptosis. Whether apoptosis occurred in myogenic cells expressing large T antigen (Tag) from SV40 upon differentiation was unknown. Human muscle satellite cells were transfected with two different constructs, containing either an origin-defective SV40 genome or Tag under vimentin promoter control. When differentiation was triggered, Tag expression reduced the formation of myotubes and dead cells showing apoptotic features were present. However, the cells expressing SV40 Tag under vimentin promoter control retained their capacity to form myotubes and expressed the myofibrillar proteins as myosin heavy chain and dystrophin when Tag expression was silent. Their apoptotic rate was similar to that of untransfected cells. The observation that apoptosis can be prevented by the down-regulation of Tag suggests that the programmed cell death induced in transformed cells can be reversed, and confirms the regulatory efficiency of the human vimentin promoter. 相似文献
12.
The novel polyamine derivatives sulphonamido oxa-spermine (oxa-Spm) and sulphonamido oxa-spermidine (oxa-Spd) exhibited rapid
cytotoxic action towards MCF-7 human breast cancer cells with IC50 values of 4.35 and 6.47 μM, respectively, after 24-h drug exposure. Neither compound is a substrate of serum amine oxidase.
Both oxa-Spm and oxa-Spd caused cell shrinkage, as determined by phase-contrast microscopy. After incubation with 10 μM of
either compound for 8 h, the cells underwent chromatin condensation and nuclear fragmentation. However, no clear DNA ladder
was obtained by electrophoresis. The sulphonamido oxa-polyamine derivatives and especially oxa-Spd enhanced the activity of
polyamine oxidase (PAO), an enzyme capable of oxidising N1-acetylated spermine and spermidine to spermidine and putrescine, respectively, generating cytotoxic H2O2 and 3-acetamidopropanal as by-products. The intracellular polyamine content was only marginally reduced in response to drug
treatment. In conclusion, our data show that these novel sulphonamido oxa-polyamine derivatives possess high cytotoxic activity
against MCF-7 cells and indicate that induction of PAO may mediate their cytotoxicity via apoptosis.
Received 17 January 2002; received after revision 22 February 2002; accepted 22 February 2002 相似文献
13.
Hydrogen peroxide and hydroxyl radical involvement in the activation of caspase-3 in chemically induced apoptosis of HL-60 cells 总被引:3,自引:0,他引:3
Kajiwara K Ikeda K Kuroi R Hashimoto R Tokumaru S Kojo S 《Cellular and molecular life sciences : CMLS》2001,58(3):485-491
Apoptosis of HL-60 cells induced by actinomycin D, H7, or daunorubicin was shown to involve the activation of caspase-3-like
protease, 2 h after the addition of these drugs, based on microassay of enzyme activity by high-performance liquid chromatography.
Catalase and a spin trap, N-t-butyl--phenylnitrone, which effectively inhibited the apoptosis induced by these drugs, also inhibited the activation of caspase-3-like
protease. These results suggest that hydrogen peroxide and the hydroxyl radical are common mediators of caspase-3 activation
caused by these chemicals, with apparently different functional mechanisms. Based on mitochondrial activity determined by
oxygen consumption, complexes I, II, and IV were inhibited by actinomycin D. H7 inhibited complexes I and IV, 1 and 1.5 h
respectively, after the addition of the drug to HL-60 cells. Daunorubicin inhibited complex IV, 1.5 h after the addition of
the drug to HL-60 cells. Inhibition of complex IV by actinomycin D, H7, and daunorubicin were almost fully restored by the
addition of cytochrome c. The release to the cytosol of cytochrome c by these drugs was also demonstrated by Western blot
analysis. Addition of catalase inhibited the depression of complex IV activity induced by actinomycin D and H7. These observations
indicate a direct relationship between hydrogen peroxide and the release of cytochrome c during apoptosis caused by actinomycin
D, H7, and daunorubicin.
Received 24 November 2000; received after revision 2 January 2001; accepted 30 January 2001 相似文献
14.
Survival of docetaxel-resistant prostate cancer cells in vitro depends on phenotype alterations and continuity of drug exposure 总被引:4,自引:0,他引:4
Makarovskiy AN Siryaporn E Hixson DC Akerley W 《Cellular and molecular life sciences : CMLS》2002,59(7):1198-1211
We evaluated in vitro the effect of paclitaxel and docetaxel on PC-3 and DU-145 prostate cancer cell lines to understand
better the downstream events in drug-induced tumor cell death. Taxane treatments of DU-145 cells induced rapid cell death
by apoptosis, but in PC-3 cells, treatments achieved growth arrest, followed by extensive karyokinesis resulting in multinucleation,
giant-cell formation and delayed cell death. To determine if the giant multinucleated cells were able to produce proliferating
and drug-resistant survivors, we first delineated the kinetics of drug activity and cytotoxic dose range. Analysis of both
lines by colorimetric and cell viability assays demonstrated improved cytotoxicity of taxanes applied continuously. Selected
doses and schedules of docetaxel were used to induce giant multinucleated cells that gave rise to docetaxel-resistant survivors,
which remained sensitive to paclitaxel and other chemotherapeutics. Growth and morphology of the recovered clones was similar
to parental cells. The resistant phenotype of these clones determined by immunofluorescence and immunoblot was associated
with transient expression of the β-tubulin IV isoform and was independent of P-glycoprotein, bcl-2 and bcl-xL. Resistant clones
will be useful to model progression of resistance to taxanes and to identify unknown and clinically important molecular mechanisms
of cell death and resistance.
Received 15 March 2002; received after revision 25 April 2002; accepted 27 May 2002 相似文献
15.
Until recently, the expression and primary function of the cell surface receptor CD40 and its ligand CD154 were considered
restricted to B and T lymphocytes, and their interactions required for the thymus-dependent humoral response. However, current
work from several groups challenges this view of the CD40/CD154 dyad as a mere mediator of lymphocyte communication. A variety
of non-lymphocytic cell types express both receptor and ligand, including hematopoetic and non-hematopoetic cells, such as
monocytes, basophils, eosinophils, dendritic cells, fibroblasts, smooth muscle, and endothelial cells. Accordingly, ligation
of CD40 mediates a broad variety of immune and inflammatory responses, such as the expression of adhesion molecules, cytokines,
matrix-degrading enzymes, prothrombotic activities, and apoptotic mediators. Consequently, CD40 signaling has been associated
with pathogenic processes of chronic inflammatory diseases, such as autoimmune diseases, neurodegenerative disorders, graft-versus-host
disease, cancer, and atherosclerosis. This review focuses on the synthesis and structure of CD40 and outlines CD154/CD40 signaling
pathways, and emphasizes the previously unexpected importance of the CD40/CD154 receptor/ligand dyad in a spectrum of immunoregulatory
processes and prevalent human diseases.
Received 10 January 2000; revised 16 June 2000; accepted 5 July 2000
RID="†"
ID="†" Review
RID="*"
ID="*" Corresponding author. 相似文献
16.
Higgins GC Devenish RJ Beart PM Nagley P 《Cellular and molecular life sciences : CMLS》2011,68(22):3725-3740
Primary neurons undergo insult-dependent programmed cell death. We examined autophagy as a process contributing to cell death
in cortical neurons after treatment with either hydrogen peroxide (H2O2) or staurosporine. Although caspase-9 activation and cleavage of procaspase-3 were significant following staurosporine treatment,
neither was observed following H2O2 treatment, indicating a non-apoptotic death. Autophagic activity increased rapidly with H2O2, but slowly with staurosporine, as quantified by processing of endogenous LC3. Autophagic induction by both stressors increased
the abundance of fluorescent puncta formed by GFP-LC3, which could be blocked by 3-methyladenine. Significantly, such inhibition
of autophagy blocked cell death induced by H2O2 but not staurosporine. Suppression of Atg7 inhibited cell death by H2O2, but not staurosporine, whereas suppression of Beclin 1 prevented cell death by both treatments, suggesting it has a complex
role regulating both apoptosis and autophagy. We conclude that autophagic mechanisms are activated in an insult-dependent
manner and that H2O2 induces autophagic cell death. 相似文献
17.
K. von Schwarzenberg S. A. E. Held A. Schaub K. M. Brauer A. Bringmann P. Brossart 《Cellular and molecular life sciences : CMLS》2009,66(7):1295-1308
In order to analyze the effects of peroxisome proliferator-activated receptor-γ (PPARγ) activation on renal cell carcinomas
we utilized several cell lines that were treated with the high affinity PPARγ agonist, troglitazone. Incubation of RCC cells
with troglitazone resulted in reduced secretion of growth factors that was due to the inhibition of MAP kinase signaling and
reduced nuclear localized expression of relB and HIF1alpha. Interestingly, the cell lines used showed a different sensitivity
towards apoptosis induction that did not correlate with the inhibition of growth factors or expression of pro- and antiapoptotic
molecules. To overcome this resistance the cells were treated with a combination of troglitazone and the proteasome inhibitor,
bortezomib. The combination of both compounds induced apoptosis even in cells resistant to both agents alone, due to increased
induction of ER-stress and caspase-3 mediated cell death.
Received 03 September 2009; received after revision 02 February 2009; accepted 10 February 2009 相似文献
18.
R. M. H. Ravindranath M. H. Ravindranath M. C. Graves 《Cellular and molecular life sciences : CMLS》1997,53(9):750-758
IgM antibodies directed against neuronal gangliosides GM1 , GM2 , GD1a , GD1b and GT1b occur in normal individuals and their level significantly decreases with age. Patients with lower motor neuron disease (LMND)
produce high levels of these autoantibodies. AntiGM1 IgM is selectively augmented. In these patients, the CD5+ (B1) and CD5− (B2) subsets of B cells are not distinct entities
but range from those without detectable CD5 marker to those with high CD5+ expression. B1 B cells were sorted to homogeneity,
but B2 B cell cannot be isolated to homogeneity because of the presence of B1 cells with low CD5 expression. In short term
cultures both the subsets produced IgM antibodies, but the antibodies reacted better with desialylated GM1 than with GM1 . Cycloheximide (Cx) (0.35 mM) largely blocked IgM synthesis of the B1 B cells but inhibition of the B2 B cells was incomplete,
possibly due to shedding of cytophilic antibodies as well as to the presence of B1 phenotype with loss of CD5 expression.
CD5+ B cells may be involved in the production of antiglycolipid IgM.
Received 9 June 1997; received after revision 21 July 1997; accepted 28 July 1997 相似文献
19.
The role of thrombospondin-1 in apoptosis 总被引:3,自引:0,他引:3
The thrombospondins are a family of extracellular proteins that participate in cell-to-cell and cell-to-matrix communication.
They regulate cellular phenotype during tissue genesis and repair. Five family members, each representing a separate gene
product, probably exist in most vertebrate species. Like most extracellular proteins, the thrombospondins are composed of
several structural domains that are responsible for the numerous biological functions that have been described for this protein
family. Considerable progress has been made towards understanding the function of thrombospondins. The role of thrombospondin
in the process of apoptosis or programmed cell death has recently come into focus. In this review we will concentrate on the
role of thrombospondin-1 in the broad field of apoptotis research.
Received 5 December 2001; received after revision 28 March 2002; accepted 28 March 2002 相似文献
20.
Programmed cell clearance 总被引:10,自引:0,他引:10
Fadeel B 《Cellular and molecular life sciences : CMLS》2003,60(12):2575-2585
Apoptosis, a physiological process of self-annihilation, is essential during development and for the maintenance of tissue homeostasis. Considerable efforts have been made in recent years to elucidate the molecular mechanisms that govern this mode of cellular demise; however, the subsequent recognition and removal of apoptotic corpses by neighboring phagocytes has received less attention. Nevertheless, macrophage engulfment of apoptotic cells is known to be important in the remodeling of tissues, and contributes to the resolution of inflammation through the removal of effete cells prior to the release of noxious cellular constituents. Moreover, apoptotic cells are a potential source of self-antigens, and clearance of cell corpses is thought to preclude the induction of autoimmune responses. The view is thus emerging that tissue homeostasis is dependent not only on the balance between mitosis and apoptosis, but also on the rate of apoptosis versus that of cell clearance. This review aims to discuss the mechanisms and consequences of macrophage recognition and disposal of apoptotic cells, a process which will be referred to as programmed cell clearance.Received 16 April 2003; received after revision 22 May 2003; accepted 26 May 2003 相似文献