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