It allows for the programmed removal of specific cells, without harming nearby cells. Staurosporine induced apoptosis in p388d1 macrophages involves both extrinsic and intrinsic pathways article in cell biology international 339. Staurosporine induces programmed cell death in embryonic. Pdf neuroprotection signaling pathway of nerve growth. Extrinsic apoptosis pathway is one of the signal pathways which may trigger the process of programmed cell death namely cell apoptosis.
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 is a very potent inducer of apoptosis because it inhibits many different kinases. These two pathways normally coexist in l1210s cells. Apoptosis can be induced by various stimuli including dnadamaging anticancer drugs and the protein kinase inhibitor staurosporine. Untreated and sts treated hela and jurkat lysates were prepared in 1x cell extraction buffer ptr and tested the cleaved parp simplesteptm elisa. The intrinsic signaling pathway for programmed cell death involves nonreceptormediated intracellular signals, inducing activities in the mitochondria that initiate apoptosis. 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 apoptosis of melanoma by both. Staurosporine transiently increased the phosphotransferase activity of cjun nterminal kinase1 jnk1, which in turn may activate the transcriptional activity of activating protein1 ap1.
Staurosporine induces apoptosis of melanoma through caspasedependent and independent pathways. Mechanisms of staurosporine induced apoptosis in a human. In contrast with apoptosis, cleanup of cell debris by phagocytes of the immune system is generally more difficult. Since staurosporine induced apoptosis in the absence of death receptor signaling, we focused on intrinsic apoptosis pathways. Introduction to apoptosis introduction apoptosis or programmed cell death is defined as a mechanism of cellular suicide which occurs after sufficient cellular damage. Staurosporine is a potent, nonselective inhibitor of protein kinases, including protein kinase c. To confirm the role of caspase activation in staurosporineinduced apoptosis of melanoma, we treated igr3 and melrm cells with the pancaspase inhibitor, zvadfmk, 1 h before adding staurosporine at 1.
We activated the death pathway in embryonic chick cerebral hemisphere neuron e7ch cultures with staurosporine 0. Apoptosis signaling pathway apoptosis is a physiological process for cell death that is critical during aging and development. Staurosporine is a cell permeable alkaloid isolated from streptomyces staurosporeus exhibiting anticancer activity. There are two major apoptosis pathways distinguished according to whether caspases are involved or. Neuroprotection signaling pathway of nerve growth factor and brainderived neurotrophic factor against staurosporine induced apoptosis in hippocampal h197 cells. Biorbyt has an extensive range of small molecules available as research reagents to target proteins in the apoptosis pathway. Nuclear chromatin condensation, cytoplasmic shrinking, dilated endoplasmic reticulum, and membrane blebbing characterize apoptosis in general. Our study showed that staurosporine induces apoptosis in pancreatic carcinoma cells via the intrinsic signaling pathway.
The initiator caspases2, 8, 9, 10, and 12 are closely. We induced apoptosis in cultured rat hippocampal neurons by exposure to the protein kinase inhibitor staurosporine 30 nm, 24 hr. Thus, staurosporine is a suitable positive control for in vitro apoptosis tests for the pancreatic cancer cell lines patu 8988t and panc1. Apoptosis very probably plays a key part in endothelial cell loss during corneal storage in organ culture as well as hypothermic storage.
Thus, staurosporine induces apoptotic cell death through at least two redundant parallel pathways. So far, many different kinase pathways of staurosporineinduced apoptosis have been discussed for various tumor entities. It is a potent, cell permeable protein kinase c inhibitor with an ic50 of 0. There are many causes of necrosis including injury, infection, cancer. Molecular mechanism of staurosporineinduced apoptosis in. Apoptosis is a distinct form of cell death that is functionally and morphologically different from necrosis. The relationship is highlighted in figure 1, which shows an overview of the acquired images. Defects of this process play an important role in a variety of diseases. The caspasedependent apoptotic pathway was activated relatively soon after exposure to.
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. 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. Take a look at our product pages where you will find competitive pricing, lead times, and easy delivery options. Staurosporine cas 62996741 98% purity ab120056 abcam. Apoptosis is known as a physiological process of cell deletion and is also a process of programmed cell death, resulting in morphological change and dna fragmentation. Apoptosis vs necrosis necrosis is the uncontrolled pathological cell death.
Staurosporine and conventional anticancer drugs induce. Induce apoptosis with staurosporine immunochemistry. We tested the hypothesis that staurosporine would also. Staurosporine induces apoptosis through both caspase. Inhibition of the devdcleaving activity by a synthetictetrapeptide inhibitor devdcho, blocked staurosporineinduced downstream apoptotic phenotypes, suchas morphological characteristics and dna fragmentation. Furthermore, staurosporine increased the transcriptional activity of nuclear factor. Calcium and reactive oxygen species mediate staurosporine. Stimuli for the intrinsic pathway include viral infections or damage to the cell by toxins, free radicals, or radiation. Request pdf triggering of a novel intrinsic apoptosis pathway by the kinase inhibitor staurosporine. The process of apoptosis is controlled by diversity cell signals pathway and involved in regulation of cell fate death or survival. Staurosporine targets the hippo pathway to inhibit cell. Activation of caspase9 in the absence of apaf1 the protein kinase inhibitor staurosporine.
Apoptosis is a tightly controlled pattern of cell death characterized by nuclear condensation, cell shrinkage, membrane blebbing, and dna fragmentation. Triggering of a novel intrinsic apoptosis pathway by the. This highly regulated modality of cell death occurs in such a way that the dying cells contents is neatly packaged into small packets which are rapidly taken up and degraded by local macrophages without the induction of inflammation. Staurosporine is an indolocarbazole alkaloid and an organic. Staurosporine is an alkaloid isolated from the culture broth of streptomyces staurosporesa. However, the mechanisms underlying endothelial apoptosis are poorly understood. Tumor protein p53 is a nuclear transcription factor that regulates the expression of a wide variety of genes involved in apoptosis, growth arrest, or senescence in response to genotoxic or cellular stress. Apoptosis is triggered by multisignal pathways and regulated by multicomplicated extrinsic and intrinsic ligands. Apoptosis is a physiological process in which cell death is brought about through a regulated sequence of events. The hippo signaling pathway was discovered to control organ size in drosophila through regulating cell proliferation and apoptosis yu et al. 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 apoptosisinducing staurosporine figure 1.
It is most commonly used as an inducer of apoptosis in several mammalian cell types. Furthermore, activation of this novel intrinsic apoptosis pathway by staurosporine did not rely on apaf1 and apoptosome. These data suggest that staurosporineinduced apoptosis in osteoblasts may occur via activation of jnk1, caspase3like proteases, and transcriptional factors including ap1 and nf. The protein encoded by this gene is a member of the family of inhibitor of apoptosis proteins iap and contains a single copy of a baculovirus iap repeat bir as well as a ringtype zinc finger domain. Neuroprotection signaling pathway of nerve growth factor and brainderived neurotrophic factor against staurosporine induced apoptosis in.
We detected the inhibitory effect in the early stage apoptosis, but later caspase activation becomes obvious. At higher concentration 120 nm, staurosporine also inhibits other kinases such as pka, pkg. Staurosporine can induce apoptosis independently of the er stress or the mitochondrial apoptosis pathway. Protection by z vadfmk observed in l1210s cells was not lasting and unmasked a secondary process of cell death that also exhibited characteristics of apoptosis. Staurosporine induced apoptosis of endothelial cells involves activation of caspase3, and could be a useful model to study strategies of cell death inhibition. The significance of apoptosis the development and maintenance of multicellular biological systems depends on a sophisticated interplay between the cells forming the organism, it sometimes even seems to involve an altruistic behaviour of individual cells in favour of the organism as a whole. Staurosporine induces apoptosis in pancreatic carcinoma. M staurosporine triggers apoptosis in nearly 40% of the cells after 12 hours, rapidly activates caspase3, and induces parp cleavage. Apoptosis can be triggered by events both inside and outside of the cell.
Further clinical studies should analyze the impact of this finding on cancer treatment. Apoptosis inducer and protein kinase inhibitorstaurosporine. Using hydroethidinebased digital videomicroscopy, we. In the present study, we have examined the apoptosisinducing potential of staurosporine in cultured melanoma cell lines and dissected the staurosporineinduced apoptotic signaling pathway. Staurosporine is often used as a general method for inducing apoptosis 1. Staurosporine is the precursor of the novel protein kinase inhibitor midostaurin pkc412. Every normal living cell of animals, plants and even bacteria are mortal. The central regulators of apoptosis are the caspases, a family of cysteine proteases that fall into two broad categories. Staurosporineinduced apoptosis in p388d1 macrophages. Staurosporine, a nonselective protein kinase inhibitor, has been shown to induce apoptosis in several different nonneuronal cell types. Based on morphological and biochemical criteria, staurosporine induced apoptosis in a human corneal endothelial cell line has been clearly demonstrated in this study.
We induced apoptosis in murine osteoblast mc3t3e1 cells by exposure to the staurosporine. Furthermore, the fact that staurosporineinduced apoptosis in l12100 cells proceeds through a caspaseindependent pathway is not specific to. The response of a human corneal endothelial cell hcec line to staurosporine, a known inducer of apoptosis, was investigated to gain insights into the. Unlike conventional anticancer drugs, staurosporine and ucn01 induced apoptosis in a variety of tumor cells overexpressing the apoptosis inhibitors bcl2 and bclx l. Besides midostaurin, staurosporine is also used as a starting material in the commercial synthesis of k252c also called staurosporine aglycone. We report that although staurosporine activated bax and the mitochondrial caspasedependent apoptotic pathway, it also induced apoptosis of melanoma by caspaseindependent pathways. Neuroprotection signaling pathway of nerve growth factor. Staurosporine has long been used in vitro as an initiator of apoptosis in many different cell types, but the mechanism involved remains poorly understood. Apoptosis is first characterized by a change in the refractive index of the cell followed by cytoplasmic shrinkage and nuclear condensation. Antibodies, kits, and reagents for apoptosis research. Apoptosis is a unique morphological pattern of cell death which is coordinated by tightly regulated signaling pathways within dying cell.