Induction of apoptosis was completely inhibited by the broad caspase inhibitor zvad figure. Staurosporine induces apoptosis through both caspasedependent. Staurosporine sts has been viewed as a broad spectrum inhibitor of protein kinases 25. It has been well established and very widely used to promote intracellular stress induced apoptosis in cell culture models employing malignant cell lines, in which the. Apr 10, 2001 apoptosis can be induced by various stimuli including dnadamaging anticancer drugs and the protein kinase inhibitor staurosporine. Staurosporine has long been used in vitro as an initiator of apoptosis in many different cell types, but the mechanism involved remains poorly understood. Exposure of murine cortical cell cultures to 30100 nm staurosporine induced concentrationdependent selective neuronal degeneration over the following day. Molecular mechanism of staurosporineinduced apoptosis in. Mechanisms of staurosporine induced apoptosis in a human. Calcium and reactive oxygen species mediate staurosporine. Mechanism of staurosporineinduced apoptosis in murine.
Thus sts causes hepatocytes to commit to death upstream of the activation of caspases. These two pathways normally coexist in l1210s cells. Jan 28, 2019 our study showed that staurosporine induces apoptosis in pancreatic carcinoma cells via the intrinsic signaling pathway. Pretreatment with caspase inhibitor converted sts induced apoptosis of hepatocytes to necrotic cell death without significantly changing total cell death. Staurosporine, a microbial alkaloid, is a strong inhibitor of protein kinases. M staurosporine triggers apoptosis in nearly 40% of the cells after 12 hours, rapidly activates caspase3, and induces parp cleavage. Previously we studied the nature of the switch mechanism between apoptosis and necrosis and investigated the intrinsic apoptotic pathway in.
In contrast, zvadfmk pretreatment protected the cells from tnf. Therefore, the objective of this study was to determine if shigella has the ability to inhibit apoptosis in. Staurosporine induces different cell death forms in cultured. This protein kinase inhibitor belongs to a family of kinase inhibitors containing an indole carbozol chromophore and is a potent inhibitor of a number of kinases. Staurosporineinduced apoptosis in l12100 cells develops only by the caspaseindependent mechanism due to a general defect in caspase activation. We report that although staurosporine activated bax and the mitochondrial caspasedependent apoptotic pathway, it also induced apoptosis of melanoma by caspaseindependent pathways. Staurosporine and conventional anticancer drugs induce overlapping, yet distinct pathways of apoptosis and caspase activation.
The relationship is highlighted in figure 1, which shows an overview of the acquired images. Request pdf protective effects of docosahexaenoic acid in staurosporineinduced apoptosis. Further clinical studies should analyze the impact of this finding on cancer treatment. The involvement of caspases in staurosporine induced cell death was also suggested by the observation that the addition of zvadfmk, a general inhibitor of caspases, suppressed apoptosis. These data suggest that staurosporineinduced apoptosis in osteoblasts may occur via activation of jnk1, caspase3like proteases, and transcriptional factors including ap1 and nf. Staurosporine induced apoptotic death in ng10815 cells. The pretreatment with the caspase inhibitor converted stsinduced apoptosis to necrosis without significantly changing the viability of the cells. Mechanisms of staurosporine induced apoptosis in a human corneal. These data suggest strongly that hcec cell death induced by staurosporine is apoptosis. Neuroprotection signaling pathway of nerve growth factor and brainderived neurotrophic factor against staurosporine induced apoptosis in hippocampal h197 cells. Staurosporine can be used to induce positive controls. Abstract background local anesthetics, especially lidocaine, can lead to persistent cauda equina syndrome after spinal anesthesia. So far, many different kinase pathways of staurosporineinduced apoptosis have been discussed for various tumor entities. Staurosporine induces necroptotic cell death under caspase.
Staurosporineinduced apoptotic water loss is cell and. Involvement of phosphatidylinositol3 kinase pathway docosahexaenoic acid 22. Recently, lidocaine has been reported to trigger apoptosis, although the underlying mechanisms remain unknown. We tested the hypothesis that staurosporine would also induce apoptosis in central neurons. Induce apoptosis with immunochemistry technologies staurosporine, a protein kinase inhibitor.
Similar results were obtained with pretreatment with devdfmk, a. Neuroprotection signaling pathway of nerve growth factor and brainderived neurotrophic factor against staurosporine induced apoptosis in hippocampal h197igfir corrected. Typically, the broadspectrum kinase inhibitor staurosporine has been the small molecule of choice, as it requires a short time period for induction of apoptosis relative to other agents. Staurosporineinduced apoptosis in p388d1 macrophages. Exposure to dead spores induced an opposite effect. We report that although staurosporine activated bax and the mitochondrial caspasedependent apoptotic pathway, it also induced apoptosis of melanoma by. Profilin 1 potentiates apoptosis induced by staurosporine in. Besides midostaurin, staurosporine is also used as a starting material in the commercial synthesis of k252c also called staurosporine aglycone. Staurosporineinduced cell death in trypanosoma brucei and. Pdf staurosporine and conventional anticancer drugs induce. It is generally believed that the molecular events during execution of apoptosis are shared, as both anticancer drugs and staurosporine derivatives induce mitochondrial damage.
Shigella flexneri inhibits staurosporineinduced apoptosis in. Distinction between different forms of cell death is crucial in the studies of therapy induced necroptosis. Staurosporine is often used as a general method for inducing apoptosis 1. 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. Maurelli department of microbiology and immunology, f. Staurosporine induces apoptosis through both caspase. The caspasedependent apoptotic pathway was activated relatively soon after exposure to. To elucidate the pathway of lidocaine induced apoptosis, the authors used genetically modified cells with overexpression or. Agents capable of inducing intrinsic pathway mediated apoptosis are widely employed in a range of biochemical experiments. Protective effects of docosahexaenoic acid in staurosporine.
Add 1 m final concentration staurosporine eg, sigma s6942 to cell suspension e. 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. We induced apoptosis in cultured rat hippocampal neurons by exposure to the protein kinase inhibitor staurosporine 30 n m, 24 hr. Staurosporine induces apoptosis in pancreatic carcinoma cells. Staurosporineinduced apoptosis of cultured rat hippocampal. We conclude that staurosporine induced dehydration is not a universal cell response but depends on the cell type and substrate attachment and can only be judged by direct water measurements. Lidocaine induces apoptosis via the mitochondrial pathway. Doxorubicin is a dna intercalator, and is commonly used to figure 2. Staurosporineinduced apoptosis in chang liver cells is. Furthermore, staurosporine increased the transcriptional activity of nuclear factor. A small molecule that induces intrinsic pathway apoptosis. Thus, staurosporine is a suitable positive control for in vitro apoptosis tests for the pancreatic cancer cell lines patu 8988t and panc1.
In the natural biosynthetic pathway, k252c is a precursor of staurosporine. Nci staurosporine is an indolocarbazole alkaloid and an organic. These findings point to a novel pathway for caspase3 activation by sts in primary hepatocytes. Pdf staurosporine, a nonselective protein kinase inhibitor, has been shown to induce apoptosis in several different nonneuronal cell types. The effects of potassium or chloride channel blockers do not always correlate with the avd. We showed that staurosporine, depending on concentration, induces both, apoptosis as well as necroptosis. Unlike conventional anticancer drugs, staurosporine and ucn. This agent induces apoptosis by an undetermined mechanism. The intracellular signalling pathways of staurosporine triggered apoptosis are however not fully known, and depend on cell type.
Staurosporine is the precursor of the novel protein kinase inhibitor midostaurin pkc412. Introduction there are two basic pathways or mechanisms of cell death. The shigella ipab protein activates caspase 1 in macrophages, which eventually leads to apoptosis. Staurosporine induced apoptosis of endothelial cells involves activation of caspase3, and could be a useful model to study strategies of cell death inhibition. We induced apoptosis in murine osteoblast mc3t3e1 cells by exposure to the staurosporine. It is generally believed that the molecular events during. Staurosporine and conventional anticancer drugs induce. We induced apoptosis in murine osteoblast mc3t3e1 cells by exposure to the. Staurosporine induces apoptosis of melanoma by both caspase. Differential regulation of specific sphingolipids in colon. Pdf neuroprotection signaling pathway of nerve growth. Unexpected effects of staurosporine on the induction of apoptosis were the activation of caspase8, and an increase of the levels of tnf the increased tnf. Based on morphological and biochemical criteria, staurosporine induced apoptosis in a human corneal endothelial cell line has been clearly demonstrated in this study. Staurosporine is a potent, nonselective inhibitor of protein kinases, including protein kinase c.
We observed i the increased cytosolic localization of p53r273h, ii the activation of phosphorylation at ser15, iii its mitochondrial localization. Furthermore, activation of this novel intrinsic apoptosis pathway by staurosporine did not rely on apaf. An indicator of apoptosis in imagebased assays bd biosciences september 2012 page 3 results images acquired showed a distinct labeling pattern of the activated caspase3 in response to the apoptosis inducing staurosporine figure 1. Further studies showed that mutant p53r273h was involved in apoptosis induced by staurosporine sts via transcriptionindependent mitochondrial functions. However, the early cell death mechanism depending on caspase activation disguises the. These results suggest that staurosporine inducedapoptosis in chok1 cells is mediated through the cpp32caspase3like cysteine proteases. Encephalitozoon cuniculi and vittaforma corneae phylum. Cell death was induced by different concentrations of staurosporine, and modified by inhibitors of apoptosis zvadfmk and necroptosis nec 1.
Staurosporineinduced apoptosis in p388d1 macrophages involves both extrinsic and intrinsic pathways article in cell biology international 339. Staurosporine can induce apoptosis andor necroptosis in cultured astrocytes via different signalling pathways. Staurosporine induces different cell death forms in. In huh6 and hepg2 cells treatment with staurosporine induced the arrest of cells in g2m phase of cell cycle.
Inhibition of the devdcleaving activity by a synthetictetrapeptide inhibitor devdcho, blocked staurosporine induced downstream apoptotic phenotypes, suchas morphological characteristics and dna fragmentation. Apoptosis can be induced by various stimuli including dnadamaging anticancer drugs and the protein kinase inhibitor staurosporine. The main consequence of hcec apoptosis is shedding. Induction of apoptosis by staurosporine involves the inhibition of. Staurosporine is a therapeutic agent that inhibits tumor cell growth by inducing cell death via intrinsic apoptotic pathways. Jun 22, 2001 furthermore, the fact that staurosporine induced apoptosis in l12100 cells proceeds through a caspaseindependent pathway is not specific to the drug used, as caspaseindependent apoptosis can. Staurosporineinduced neuronal apoptosis sciencedirect. Staurosporine transiently increased the phosphotransferase activity of cjun nterminal kinase1 jnk1, which in turn may activate the transcriptional activity.
Triggering of a novel intrinsic apoptosis pathway by the. Induce apoptosis with staurosporine immunochemistry. Human peripheral t lymphocytes anticd2 or staurosporine in. 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. Staurosporine is a cell permeable alkaloid isolated from streptomyces staurosporeus exhibiting anticancer activity. Both species of microsporidia modulated the intrinsic apoptosis pathway. Caspase inhibition prevents staurosporineinduced apoptosis. Staurosporine also inhibits topoisomerase ii directly by interaction with the atp binding site, augments pma induced ornithine decarboxylase, and activates a bcl2regulated apoptosis pathway. Different forms of a cell death were detected using flow cytometry. Staurosporine, a nonselective protein kinase inhibitor, has been shown to induce apoptosis in several different nonneuronal cell types. Shigella flexneri is a facultative intracellular organism that causes bacillary dysentery.
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