Further studies showed that mutant p53r273h was involved in apoptosis induced by staurosporine sts via transcriptionindependent mitochondrial functions. Thus, staurosporine is a suitable positive control for in vitro apoptosis tests for the pancreatic cancer cell lines patu 8988t and panc1. Staurosporineinduced apoptosis of cultured rat hippocampal. Dec 04, 2020 staurosporine induces apoptosis in the jurkat subclone jm319 resistant to anticancer drug and cd95 induced apoptosis. Staurosporine induces apoptosis of melanoma by both. Mitochondrial outer membrane permeability change and. Staurosporineinduced apoptosis is independent of p16 and.
Staurosporine induces platelet apoptosis through p38. Sep 01, 2009 an agent frequently used for the induction of apoptosis is staurosporine. Staurosporine induces apoptosis of melanoma by both caspase. Although the mitochondrial apoptotic pathway was activated by staurosporine. Staurosporine induces apoptosis in pancreatic carcinoma cells. We induced apoptosis in murine osteoblast mc3t3e1 cells by exposure to the. In this study, staurosporineinduced apoptosis model was used. Conclusions these results suggest that fkbp51 protects 661w cell cultures from apoptosis induced by staurosporine.
The mechanism of sts induced apo ptosis is still unknown but it involves alteration of the ph osphorylation state, alterations related to cell cycle con. These results indicate that, in cerebrocortical cultures, excessive nmda receptor activation precipitates neuronal apoptosis by means of mitochondrial dysfunction, whereas staurosporine utilizes a distinct pathway. Shigella flexneri inhibits staurosporineinduced apoptosis in. Bclxl were resistant to staurosporineinduced apoptosis as assessed by dna. However, the proapoptotic role of gsk3 is still unclear. Shigella flexneri is a facultative intracellular organism that causes bacillary dysentery. Staurosporineinduced apoptosis is independent of p16 and p21. We report that although staurosporine activated bax and the mitochondrial caspasedependent apoptotic pathway, it also induced apoptosis of melanoma by caspaseindependent pathways. Staurosporine can induce death of hela cells via a cytochrome ccaspase. Here, we show the involvement of gsk3 in ceramide induced mitochondrial apoptosis. These results suggest that isoflurane may induce apoptosis through bcl2 family proteins and rosassociated mitochondrial pathway of apoptosis. Laboratory science mechanisms of staurosporine induced. Check cells to determine if cells have begun to apoptose. Therefore, the objective of this study was to determine if shigella has the ability to inhibit apoptosis in.
Trail induces necroptosis in the presence of caspase inhibitor. The effect of an hiv1 viral protease inhibitor on staurosporine. Pdf fkbp51 protects 661w cell culture from staurosporine. Staurosporineinduced cell death in trypanosoma brucei and. Mechanisms of staurosporine induced apoptosis in a human.
Triggering of a novel intrinsic apoptosis pathway by the. Molecular mechanism of staurosporineinduced apoptosis in. Sts is a generally accepted inducer of intrinsic apoptotic pathway and it is a wide spectrum inhibitor of protein kinases 31. Human peripheral t lymphocytes anticd2 or staurosporine in. The caspasedependent apoptotic pathway was activated relatively soon after exposure to. Staurosporine, a microbial alkaloid, is a strong inhibitor of protein kinases. Furthermore, activation of this novel intrinsic apoptosis pathway by staurosporine did not rely on apaf1 and apoptosome. Staurosporine induces platelet apoptosis through p38 mitogen. Staurosporine resistance in inflammatory neutrophils is. Treatment with the antioxidant 6atocopherol 100 mm or the superoxide dismutasemimetic manganese tetrakis 4benzoyl acid porphyrin 1 mm signi. Staurosporine is a therapeutic agent that inhibits tumor cell growth by inducing cell death via intrinsic apoptotic pathways. General protocols for inducing apoptosis in cells apoptosis may be induced in experimental systems through a variety of methods.
Jurkat jm319 subclone, while crossresistant to anticd95 and anticancer drug induced apoptosis, is sensitive to staurosporine discussion by continuous culture in the presence of anticd95 we generated a jurkat cell clone, called jm319, which was staurosporine is a broadspectrum protein kinase completely resistant towards cd95mediated apopto inhibitor that has been used extensively to induce sis. A small molecule that induces intrinsic pathway apoptosis with. Dose response with anticd95 a, etoposide b, daunorubicin c and. The shigella ipab protein activates caspase 1 in macrophages, which eventually leads to apoptosis. Jan 28, 2019 staurosporine is a very potent inducer of apoptosis because it inhibits many different kinases. Pten augments staurosporineinduced apoptosis in ptennull. Mar 29, 2000 furthermore, vraf blocks apoptosis induced by staurosporine. Estvez ag, spear n, manual sm, radi r, henderson ce, barbeito l. Triggering of a novel intrinsic apoptosis pathway by the kinase. Similar results were obtained with pretreatment with devdfmk, a. Staurosporine antibiotic am2282 or sts is a natural product originally isolated in 1977 from.
Cycloheximide induces apoptosis through a pathway involving fak proteolysis without dephosphorylation. In contrast, zvadfmk pretreatment protected the cells from tnf. Beyond that, staurosporine represents one of the most potent apoptotic stimuli and induces apoptosis with similar rapid kinetics as death receptor. Staurosporine is a very potent inducer of apoptosis because it inhibits many different kinases. Unlike conventional anticancer drugs, staurosporine and ucn. Caspase dependent pathway caspasedependent apoptosis is the classic programmed cell death pathway, the capase8. Cytokine regulation of apoptosisinduced apoptosis and. Tdp43 plays a key role in cell cycle and apoptosis. Mechanism of staurosporineinduced apoptosis in murine hepatocytes. Although important regulators of the cell death pathway have been identified.
It has been found that one way in which staurosporine induces apoptosis is by activating. Staurosporine induced neuronal apoptosis was significantly reduced by 24h pretreatments with the protein synthesis inhibitor cycloheximide i pm. Dids attenuates staurosporineinduced cardiomyocyte apoptosis. Further clinical studies should analyze the impact of this finding on cancer treatment. May 01, 2002 the pretreatment with the caspase inhibitor converted sts induced apoptosis to necrosis without significantly changing the viability of the cells. Epidermal growth factor egf suppresses staurosporine. Staurosporine induces endothelial cell apoptosis via focal. Maurelli department of microbiology and immunology, f. We induced apoptosis in cultured rat hippocampal neurons by exposure to the protein kinase inhibitor staurosporine 30 nm, 24 hr. Zhao l, lu g, zhao q, zhang m, chen m, zhang j, dai k. We report that although staurosporine activated bax and the mitochondrial caspasedependent apoptotic pathway, it also induced apoptosis of melanoma by. Gsk3 acts downstream of pp2a and the pi 3kinase akt.
The mitochondria, as the crosstalk organelles, can connect the different apoptosis pathway. Arising studies over the past two decades have defined the core components and regulation mechanisms of the pathway in both drosophila and mammals. Staurosporine induces plate let apoptosis through p38 mitogenactivated proteinkinase signaling pathway. We report in this study that primary cultured mouse hepatocytes are less sensitive to sts. So far, many different kinase pathways of staurosporine induced apoptosis have been discussed for various tumor entities. Staurosporine has long been used in vitro as an initiator of apoptosis in many different cell types, but the mechanism involved remains poorly understood. Id, two agents that have been reported to inhibit neuronal apoptosis caused by withdrawal of trophic factors fig. Pdf staurosporine and conventional anticancer drugs. Yet the mechanisms leading to apoptosis in shsy5y neuroblastoma cells following exposure to staurosporine and h7 are not clearly understood. We report that staurosporine induced relatively high levels of apoptosis in the majority of melanoma cell lines through both caspasedependent and independent pathways. Jun 21, 2019 in this study, staurosporine induced apoptosis model was used in order to compare lowcost and highcost annexin v detection kits from three different producers. This alkaloid originally isolated from streptomyces staurosporeus has been described as an inhibitor of protein kinases tamaoki and nakano, 1990. In the present study, we have examined the apoptosis inducing potential of staurosporine in cultured melanoma cell lines and dissected the staurosporine induced apoptotic signaling pathway. Ic or with mimosine, a cell cycle inhibitor at the g 1 is transition 100 pm.
Staurosporine induces apoptosis in pancreatic carcinoma. Staurosporine sts, a microbial alkaloid and potent pkc inhibitor,has become one of the most promising anticancer drugs. Activation of extrinsic or intrinsic apoptosis of vsmcs. Unlike conventional anticancer drugs, staurosporine and ucn01 induced apoptosis in a variety of tumor cells overexpressing the apoptosis inhibitors bcl2 and bclx l. Induction of apoptosis by staurosporine involves the inhibition of. However, prb was dephosphorylated and cdc2 was inhibited at both the low and the high concentrations of. Profilin 1 potentiates apoptosis induced by staurosporine in. In contrast to the cell lines, no apparent release of cytochrome c or loss of mitochondrial transmembrane potential was detected in primary hepatocytes undergoing stsinduced apoptosis. Thus sts causes hepatocytes to commit to death upstream of the activation of caspases. Mechanism of staurosporineinduced apoptosis in murine. Staurosporineinduced apoptosis involves the activation of mitochondrial caspase takahashi et al.
We previously reported that ceramide induced caspase2 and. Our study showed that staurosporine induces apoptosis in pancreatic carcinoma cells via the intrinsic signaling pathway. Ceramide induced gsk3 activation via protein dephosphorylation at serine 9. To identify pathways and cytokines released when vsmcs undergo apoptosis, we treated mouse vsmcs with an agonistic antibody to the death receptor cd95fas and 10 nm cycloheximide.
These findings point to a novel pathway for caspase3 activation by sts in primary hepatocytes. Profilin 1 potentiates apoptosis induced by staurosporine. Differential regulation of specific sphingolipids in colon. Id, two agents that have been reported to inhibit neuronal apoptosis caused by withdrawal of trophic factors.
Jul 22, 2020 the extrinsic pathway of apoptosis is inhibited by a protein called cflip which will bind to fadd and caspase8, rendering them ineffective. Staurosporine cell death is inhibited by bcl2, involves the release of cytochrome c and is associated with caspase activation. Apoptosis definition, pathways, assay, examples vs necrosis. Finally, the anesthetic desflurane does not induce activation of mitochondrial pathway of apoptosis. While caspases play a determining role in early apoptotic responses, aif may be involved in apoptotic execution at late stages after staurosporine treatment. May 23, 2000 additionally, staurosporine induced, but not nmda induced, apoptosis was associated with activation of caspase8. Briefly, two human cell lines kg1 and nkt were treated with staurosporine for 1, 3 and 6 hours and then evaluated for the apoptosis induction using 3 different detection kits by.
Staurosporine is a microbial alkaloid and a strong broad spectrum protein kinase inhibitor which has been shown to induce apoptosis in uni and multicellular organisms 23 27. We also found that st reduced akt phosphorylation as well as its activity and this reduction is partially prevented by dha enrichment. Apoptosis of cultured human endothelial cells was induced by the mycotoxin staurosporine, which has been shown to induce apoptosis in a wide variety of cell types. We observed i the increased cytosolic localization of p53r273h, ii the activation of phosphorylation at ser15, iii its mitochondrial localization. The mitochondrial pathway of anesthetic isofluraneinduced. Staurosporine sts induces apoptosis in various cell lines. Pdf staurosporine and conventional anticancer drugs induce. Pdf staurosporine induces apoptosis of melanoma by both. This evidence implies that the survival signal of egf is mediated via the rafmekerk pathway but not the pi3k pathway. The akt signaling pathways were investigated during p. Sb203580 reduced staurosporine induced phosphorylation of the p38mapk target mk2 while sp600125 reduced staurosporine induced phosphorylation of cjun with also some reduction of total cjun, most likely from autoactivationsupplemental fig.
Another mechanism of apoptosis regulation in the extrinsic pathway involves a protein called toso, which blocks fas induced apoptosis in t cells by inhibition of caspase8 activation. Staurosporine induces necroptotic cell death under caspase. Mitochondrial and extramitochondrial apoptotic signaling. We found that apoptosis induced by staurosporine st, a general kinase inhibitor which has been shown to induce apoptosis in various systems fukuoka et al. We report in this study that primary cultured mouse hepatocytes are less sensitive to sts compared with jurkat cells and huh7 cells. Dec 28, 2000 staurosporine and 15isoquinolinesulfonyl2methylpiperazine h7 are nonspecific protein kinase inhibitors and wellestablished proapoptotic agents. Jan 10, 2003 143b osteosarcoma cells induced to undergo apoptosis by exposure to 1.
Whereas extrinsic apoptosis induction by the activation of fas death receptor on inflammatory neutrophils was accompanied by caspase and proteasomemediated myeloid cell leukemia 1 degradation, intrinsic apoptosis induction by staurosporine led to a significant stabiliza. This rate was measured in intact cells suspended in buffered saline solution. Therefore, in immortalized mesangial cells, staurosporineinduced apoptosis was not. Fk506 treatment significantly protected 661w neuronal cultures from staurosporine induced apoptosis. Jan 01, 2009 a second objective was to investigate the participation of a novel caspase3 pathway in stp induced human colonic cell apoptosis given that stp induced hepatic apoptosis is mediated by a caspase3 pathway which is independent of mitochondrial cytochrome c release and caspase9 or caspase8 activation. Staurosporine induces apoptosis in the jurkat subclone jm319 resistant to anticancer drug and cd95 induced apoptosis. Protective effects of docosahexaenoic acid in staurosporine. To identify the effect of staurosporine in pancreatic and colorectal carcinoma cells and its apoptosis inducing signaling pathway. However, the mechanisms that activate initiator caspases leading to. It has been found that one way in which staurosporine induces apoptosis is by activating caspase3. At lower concentration, depending on the cell type, staurosporine induces specific cell cycle effects arresting cells either in g 1 or in g 2 phase of the cell cycle. Staurosporine targets the hippo pathway to inhibit cell. Staurosporine and conventional anticancer drugs induce.
Pdf staurosporine, a nonselective protein kinase inhibitor, has been shown to induce apoptosis in several different nonneuronal cell types. The hippo signaling pathway was discovered to control organ size in drosophila through regulating cell proliferation and apoptosis yu et al. Sts effectively inducesapoptosis in many nucleated cells. Porphyromonas gingivalis inhibits mitochondrioninduced. Molecular ordering of the cell death pathway journal of biological. Among numerous other experiments, staurosporine has been.
Mar 28, 2002 staurosporine is a potent apoptosis inducer, but its mechanism remains to be clarified. Doxorubicin is a dna intercalator, and is commonly used to figure 2. Mechanisms of staurosporine induced apoptosis in a human corneal. Shigella flexneri inhibits staurosporineinduced apoptosis. Pdf staurosporine induced apoptosis rapidly downregulates. The specific apoptotic nature of staurosporine induced hcec death was confirmed. The principal mediators for the apoptotic pathway are a class of protease.
We investigated the involvement of pten in staurosporine induced apoptosis. Staurosporineinduced apoptosis in p388d1 macrophages. Pretreatment with caspase inhibitor converted sts induced apoptosis of hepatocytes to necrotic cell death without significantly changing total cell death. These studies show that fak dephosphorylation and focal adhesion disassembly are very early events mediating the onset of staurosporine induced endothelial cell apoptosis and are dissociated from fak proteolysis. The isoflurane induced ros accumulation can be attenuated by the intracellular calcium chelator bapta. In contrast to the cell lines, no apparent release of cytochrome c or loss of mitochondrial transmembrane.
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