Proportions of Mitotic and Apoptotic Cells In A Range of Untreated Experimental Tumours

Sarraf, Catherine; Bowen, I. D.

doi:10.1111/j.1365-2184.1988.tb00770.x

Cited by 56 publications

(54 citation statements)

References 9 publications

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“…The dysregulation of apoptotic cell death may be involved in the pathogenesis of a variety of human diseases, including cancers, autoimmune diseases, neurodegenerative disorders, and viral infection (3)(4)(5)(6). Growing evidence suggests that although apoptotic stimuli vary from cell to cell, there seems to be a basic biochemical mechanism underlying this process, which involves the activation of a family of Cys-dependent, Asp-specific proteases known as the caspases (7)(8)(9).…”

mentioning

confidence: 99%

Caspase Activation of Mammalian Sterile 20-like Kinase 3 (Mst3)

Huang

Hsu

et al. 2002

Journal of Biological Chemistry

110

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Mammalian Sterile 20-like kinase 3 (Mst3), the physiological functions of which are unknown, is a member of the germinal center kinase-III family. It contains a conserved kinase domain at its NH 2 terminus, whereas there is a regulatory domain at its COOH terminus. In this study we demonstrate that endogenous Mst3 is specifically cleaved when Jurkat cells were treated with anti-Fas antibody or staurosporine and that this cleavage is inhibited by the caspase inhibitor, Ac-DEVD-CHO. Using apoptotic Jurkat cell extracts and recombinant caspases, we mapped the caspase cleavage site, AETD 313 , which is at the junction of the NH 2 -terminal kinase domain and the COOH-terminal regulatory domain. Caspase-mediated cleavage of Mst3 activates its intrinsic kinase activity, suggesting that the COOH-terminal domain of Mst3 negatively regulates the kinase domain. Furthermore, proteolytic removal of the Mst3 COOH-terminal domain by caspases promotes nuclear translocation. Ectopic expression of either wild-type or COOH-terminal truncated Mst3 in cells results in DNA fragmentation and morphological changes characteristic of apoptosis. By contrast, no such changes were exhibited for catalytically inactive Mst3, implicating the involvement of Mst3 kinase activity for mediation of these effects. Collectively, these results support the notion that caspase-mediated proteolytic activation of Mst3 contributes to apoptosis.

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mentioning

confidence: 99%

Caspase Activation of Mammalian Sterile 20-like Kinase 3 (Mst3)

Huang

Hsu

et al. 2002

Journal of Biological Chemistry

110

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“…Though conspicuous histologically, necrosis does not satisfactorily account for most of the losses predicted on the basis of CLF. 13,14 It is therefore likely that apoptosis, which exists in all tumors, 15,16 is responsible for an important part of cell death in neoplasia. Due to the rapidity of the phenomenon (2-5 min for cell condensation and fragmentation, 3 h for apoptotic-body phagocytosis and digestion), strong ACD-induced cell losses are often based on less than 5% of simultaneously visible apoptotic cells within a tissue.…”

Section: Introductionmentioning

confidence: 99%

Cell death and cancer: replacement of apoptotic genes and inactivation of death suppressor genes in therapy

et al. 1998

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“…One hour after injection of BrdUrd the vast majority of S-phase cells would still be in this phase and thus susceptible to the cytotoxic agents, while 4 h after administration of the antimetabolites was sufficient time to monitor their fate. As expected, BrdUrd negative apoptotic cells were sometimes seen in treated tumours due to the low spontaneous level of apoptosis (Sarraf & Bowen, 1988), conversely, particularly at the lower dose levels of the drugs, some cells (probably those that had been at the extreme end of S-phase when BrdUrd was administered) were able to survive in S-phase without being killed (Benton & Alison, 1984), and thus be labelled without being apoptotic.…”

Section: Discussionmentioning

confidence: 80%

Bromodeoxyuridine-labelled apoptosis after treatment with antimetabolites in two murine tumours and in small intestinal crypts

Sarraf

Ansari²,

Conway³

et al. 1993

Br J Cancer

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Summary Antimetabolites are S-phase specific anticancer drugs. Administration of bromodeoxyuridine (BrdUrd) to tumour bearing mice was followed by treatment with cytosine arabinoside or hydroxyurea. Anti-BrdUrd immunocytochemistry visualised susceptible tumour and intestinal crypt cells at electron microscope level, showing unequivocally that cells that were in S-phase at the time of administration of the drugs subsequently died by apoptosis.Antimetabolites are commonly used anticancer drugs that produce their cytotoxicity by targeting tumour cells in the S-phase of the cell cycle. Ijiri and Potten (1987) found that a wide range of chemotherapeutic agents caused cell death in intestinal crypts and Anilkumar et al. (1992) determined by electron microscope (EM) examination that this cell death in the mouse, when treated with cytosine arabinoside (Ara-C) or hydroxyurea (HU), was through apoptosis rather than necrosis. In tumours, the precise mode of cell death after chemotherapy is important, as apoptosis is considered as an active, gene-directed process which in some circumstances it might be possible to selectively enhance (Itoh et al., 1991). By injecting BrdUrd -a specific precursor of DNA, followed by the antimetabolite, and subsequently visualising the former by immunocytochemistry, S-phase cells would be identified then tracked to their death. Here, the nature of this cell death has been examined in two transplantable murine tumours, a sarcoma, SaF, and a carcinoma, CaX, and in murine small intestinal crypts, in response to treatment with Ara-C or HU. Materials and methods TissuesAll experiments were performed in male CBA mice bearing either SaF or CaX, when the tumours were 1 cm in diameter. Duodenal samples were obtained from the same animals. Eight animals bearing SaF and eight bearing CaX were treated with Ara-C and similar groups were treated with HU.Each tumour-bearing group also contained two control animals which received saline in the place of Ara-C or HU.Flash labelling with BrdUrd and subsequent cytotoxic drug treatment The method of Sarraf and Alison (1993) was used that ultimately demonstrated S-phase cells that subsequently died by apoptosis as determined by EM. BrdUrd was administered ip to all animals at a dose of 50 mg kg-', to identify S-phase cells at the time of drug administration. One hour later, Ara-C was given, ip, to four animals bearing SaF and four bearing CaX at 1000 mg kg', 'high dose', and to the other four of each at the 'low dose' of 100 mg kg-'. At each of these dose levels, two animals were killed 2 h later, and two were killed 4 h later. Also 1 h post BrdUrd, HU was given, ip, to four animals bearing SaF and four animals bearing CaX, at 1500 mg kg-' 'high dose' and to four bearing SaF and four bearing CaX at 150 mg kg-' 'low dose'. At each of these dose levels, once more, two animals were killed 2 h later, and two were killed 4 h later. Thus, all cells that had been in the S-phase at the time of administration of BrdUrd would be demonstrable by immunolabelling, and thei...

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Proportions of Mitotic and Apoptotic Cells In A Range of Untreated Experimental Tumours

Cited by 56 publications

References 9 publications

Caspase Activation of Mammalian Sterile 20-like Kinase 3 (Mst3)

Caspase Activation of Mammalian Sterile 20-like Kinase 3 (Mst3)

Cell death and cancer: replacement of apoptotic genes and inactivation of death suppressor genes in therapy

Bromodeoxyuridine-labelled apoptosis after treatment with antimetabolites in two murine tumours and in small intestinal crypts

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