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Biomedical subjects

I R Radford

Publications and source records attributed to I R Radford.

At least 37 records · Page 2Linked to original sources
PubMed1994

Radiation response of mouse lymphoid and myeloid cell lines. Part II. Apoptotic death is shown by all lines examined.

The mode of death induced by gamma-irradiation in a panel of 10 mouse lymphoid or myeloid cell lines was examined. Four of these lines were known to lose viability (membrane integrity) rapidly after irradiation, whilst the others were known to lose viability considerably more slowly. However, based on the criteria of morphology and DNA degradation pattern, all 10 lines showed apoptotic death. The occurrence of apoptosis after irradiation in rapid-dying lymphoid cell lines was consistent with published results, whilst the demonstration of apoptosis in slow-dying lines was unexpected. Cells of the slow-dying lymphoid lines underwent one or more mitoses prior to death, a feature also reported for fibroblastoid cell lines. However, the occurrence of radiation-induced necrosis in fibroblasts suggests that the pathways leading to 'mitotic death' differ between fibroblastoid and lymphoid cell lines.

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PubMed1994

Radiation response of mouse lymphoid and myeloid cell lines. Part III. Different signals can lead to apoptosis and may influence sensitivity to killing by DNA double-strand breakage.

We have examined the timing of gamma-irradiation-induced death in relation to cell cycle progression using a panel of mouse lymphoid or myeloid cell lines. Death was found to occur immediately after irradiation ('rapid interphase' death), or after arrest in G2 phase ('delayed interphase' death). In part II of this series of papers we demonstrated the occurrence of radiation-induced apoptosis in all these cell lines. This suggests that different signals can lead to apoptosis in these cell lines. DNA double-strand breakage appeared to directly stimulate the destruction of cell lines susceptible to rapid interphase death, whilst the signal for delayed interphase and mitotic death, whilst the signal for delayed interphase and mitotic death appeared to be chromosomal aberrations. Several of the cell lines showed different timing of death dependent upon the radiation dose used. These differences in the timing of radiation-induced death are shown to be useful indicators of the relative radiosensitivity of haematopoietic cell lines.

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PubMed1994

Phorbol esters can protect mouse pre-T cell lines from radiation-induced rapid interphase apoptosis.

Protein kinase C stimulators were found to increase the radioresistance of the mouse pre-T cell-derived line ST4. Increased resistance to gamma-ray-induced killing could be produced by addition of 10 nM phorbol 12-myristate 13-acetate (PMA) to ST4 cultures either immediately before or up to 2 h after irradiation. Following PMA treatment, ST4 changed from a cell line that underwent rapid interphase apoptosis (i.e. DNA degradation and morphology characteristic of apoptosis were evident 2-3 h after irradiation) to a line that continued to cycle after irradiation and began to die by apoptosis after completing mitosis. Associated with these PMA-induced changes, the D0 of ST4 cells increased from 7.7 +/- 0.7 to 18.8 +/- 2.7 125I decays. Another mouse pre-T cell-derived line, ST1, which is susceptible to radiation-induced rapid interphase apoptosis, also showed radioprotection after PMA treatment. In contrast, PMA increased the radiosensitivity of the pre-T cell-derived W7 line, which undergoes radiation-induced delayed interphase apoptosis (i.e. death following blockage in G2 phase). PMA had no effect on the radiosensitivity of a pre-B cell-derived line, A8, which undergoes rapid interphase apoptosis, and on a pre-T cell-derived line, W22, which undergoes apoptosis after mitosis. These results suggest that the radiomodifying ability of PMA treatment is dependent upon the cell death pathway induced by irradiation and upon the cell lineage.

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PubMed1994

p53 status, DNA double-strand break repair proficiency, and radiation response of mouse lymphoid and myeloid cell lines.

The p53 status of a panel of 10 mouse lymphoid or myeloid cell lines was determined by immunoprecipitation with mutant- and wild-type-specific antibodies and was compared with the radiation response of the lines. The more rapidly dying cell lines all contained p53 displaying the wild-type epitope. By contrast, four of six more slowly dying cell lines contained either no or mutant p53 protein. It was of interest that radiation-induced apoptosis occurred, albeit at a considerable time after irradiation, in cells ostensibly lacking p53 protein. DNA double-strand break (dsb) repair was examined in both a rapidly and more slowly dying cell line. The rapidly dying cell line was capable of DNA dsb rejoining, however this repair was interrupted by postirradiation DNA degradation.

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PubMed1991

Mouse lymphoma cells that undergo interphase death show markedly increased sensitivity to radiation-induced DNA double-strand breakage as compared with cells that undergo mitotic death.

The relationship between radiation-induced DNA double-strand breakage (dsb) and reproductive death (clonogenicity) for two mouse lymphoma cell lines was compared with that for the fibroblast-like hamster cell line V79. One of the lymphoma lines (STRij-4-2.2), which undergoes rapid disintegration following cytotoxic insult, showed extreme sensitivity to gamma-ray or DNA-associated 125I decay-induced DNA dsb (7 +/- 1 125I decays per clonogenic lethal event). Surprisingly, the other lymphoma line (WEHI-22.1), which does not undergo rapid disintegration, was also much more sensitive to DNA dsb than were V79 cells (17 +/- 1 versus 61 +/- 2 125I decays per clonogenic lethal event). Ultrastructure, DNA degradation, and flow cytometric cell cycle data suggested that both lymphoma cell lines may undergo interphase death, but that the induction of this process in WEHI-22.1 may depend upon blockage in the G2 phase. It is concluded that there are marked differences between the radiation responses of lymphoma and fibroblast lines, that there may be different forms of radiation-induced interphase death, and that the low number of DNA dsb required to produce a clonogenic lethal event in cells undergoing interphase death could explain the radiosensitivity of organs such as ovary, testis and thymus.

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PubMed1989

Fliv-1: a surface membrane differentiation antigen of fetal rat hepatocytes.

A monoclonal antibody of IgG1 subclass was raised against 15 day fetal hepatocytes. The antibody appears to recognize a surface membrane-associated fetal hepatocyte antigen. Preliminary experiments suggest that the antibody recognizes a protein epitope. The tissue distribution of the antigen, designated Fliv-1, was studied by immunoperoxidase staining on whole body sections from fetal rats of different gestational ages. In 15 day fetal rats the antigen was detected on Rathke's pouch, brain infundibulum, tongue, liver, pancreatic acini, adrenal gland, vertebral myotome, and some developing muscle tissue. All normal adult tissues that were examined, two rat hepatoma cell lines, and hepatomas induced in rats by a single injection of diethylnitrosamine, were negative for the Fliv-1 antigen. On the basis of its tissue distribution, it seems unlikely that Fliv-1 is a previously described fetal hepatocyte marker.

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PubMed1988

Aphidicolin synchronization of mouse L cells perturbs the relationship between cell killing and DNA double-strand breakage after X-irradiation.

Mouse L cells were synchronized in early S-phase by two 12 h incubations in medium containing aphidicolin (2 micrograms/ml), separated by 8 h in drug-free medium. The relationship between X-ray-induced cell killing and DNA double-strand breakage was then examined for cells that had entered S-phase, G2-phase, mitosis, and G1-phase following release from aphidicolin and was compared to the response of asynchronous cultures. Aphidicolin-synchronized cells showed cycle phase-dependent changes in their dose-responses for both killing and DNA dsb. However, on the basis of the level of DNA dsb per unit length of DNA required to produce a lethal lesion, aphidicolin-synchronized cells were more sensitive to X-rays than were asynchronous cultures. This sensitivity peaked 2 h after release from aphidicolin treatment and then progressively declined towards the asynchronous culture value. It is argued that these results are due to deregulation of the temporal order of DNA replication following aphidicolin treatment, and can be incorporated into the critical DNA target size model (Radford, Hodgson, and Matthews, in preparation) by postulating that the targets for radiation action in mammalian cells are DNA-associated with potentially transcriptionally active proto-oncogenes or constitutive fragile sites.

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PubMed1988

The dose-response for low-LET radiation-induced DNA double-strand breakage: methods of measurement and implications for radiation action models.

There is considerable controversy over the form of the dose-response for DNA double-strand breakage (dsb) induction in mammalian cells by low-LET type radiation. This controversy centres on the techniques used for measuring DNA dsb. The applications and shortcomings of the four major techniques for estimating DNA size--sedimentation, viscoelastometry, electrophoresis, and non-denaturing filter elution--are examined. In particular, the criticisms of the results obtained using the non-denaturing filter elution technique, which have suggested that the DNA dsb dose-response is non-linear, are discussed. It is concluded that these results may require a re-evaluation of the basic assumptions of many radiation action models.

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PubMed1988

Critical DNA target size model of ionizing radiation-induced mammalian cell death.

A new model of mammalian cell killing by ionizing radiation is presented. This model, termed the critical DNA target size model, postulates that DNA double-strand breakage is the critical radiation-induced lesion and that the dose-response for such breakage can be non-linear due to the action of a saturable chemical repair process. DNA double-strand breakage occurring within critical targets (proto-oncogene- or common fragile site-associated sequences) is postulated to initiate recombination events with undamaged sequences, leading to chromosomal aberrations. The subsequent loss of acentric fragments at mitosis is postulated to prevent the continuity of the genome and to produce cell death by the induction of chromatin structural changes. Experimental evidence contrary to other radiation action models is examined, and the hypotheses of the model are justified.

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PubMed1987

Effect of ploidy on the response of V79 cells to ionizing radiation.

The responses of diploid, tetraploid and near-hexaploid V79 cells to X-irradiation or DNA-associated 125I-decay were compared. When cell killing, following X-irradiation, was plotted against the induced level of DNA double-strand breakage (dsb) per unit length of DNA, there was no significant difference between the relationships for each cell line. This suggested that the number of X-ray-induced DNA dsb per cell required to produce a lethal lesion was proportional to ploidy. Consistent with the X-ray results, tetraploid cells required 121 +/- 4 and diploid cells 60 +/- 1 125I-decays to produce a lethal lesion. However, the hexaploid cells deviated from this relationship and required 137 +/- 5 decays. The relationship between relative elution and 125I decays/cell reflected cellular DNA content. It is concluded that current models of radiation action are unable to explain these findings satisfactorily.

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PubMed1987

Effect of cell-cycle position and dose on the kinetics of DNA double-strand breakage repair in X-irradiated Chinese hamster cells.

The kinetics of DNA double-strand breakage (d.s.b.) repair in X-irradiated Chinese hamster V79 cells were found to be affected by cell-cycle position. In mitotic cells, the repair kinetics were monophasic with a half-time value of about 32 min, whilst in G1, S, or asynchronous cultures, the kinetics were biphasic with half-time values of around 2.7 and 27 min. The repair of DNA single-strand breakage (s.s.b) was also shown to be slower in mitotic than in interphase cells. The DNA d.s.b. repair system, in both mitotic and interphase cells, showed no evidence of saturation within the X-ray dose range covered. The implications of these findings for the mechanism of DNA d.s.b. repair and for models of ionizing radiation action are discussed.

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PubMed1987

Use of 'nuclear monolayers' to identify factors influencing DNA double-strand breakage by X-rays.

Nuclear monolayers, prepared by treatment of mammalian cells with non-ionic detergents, showed increased sensitivity to X-ray-induced DNA double-strand breakage (dsb), as compared with intact cells, due to a decrease in the low-dose 'shoulder'. The DNA dsb dose-response shoulder could be restored by irradiating nuclei in the presence of sulphydryl compounds. However, the ineffectiveness of glutathione, when used at near cellular levels, in restoring the shoulder, suggested a possible role for protein sulphydryls in the radiation response of intact cells.

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PubMed1986

Evidence for a general relationship between the induced level of DNA double-strand breakage and cell-killing after X-irradiation of mammalian cells.

X-ray-induced DNA double-strand breakage (dsb) and lethal lesion induction have been examined using normal and transformed fibroblasts of rodent and human origin. DNA dsb induction varied markedly among the different cell types and was found to reflect radiosensitivity. Linear relationships were found between DNA dsb and lethal lesion induction which were not significantly different for each cell type, suggesting a comparable probability of conversion of DNA dsb into a lethal lesion. The implications of these findings for models of cell survival are discussed.

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PubMed1986

Effect of radiomodifying agents on the ratios of X-ray-induced lesions in cellular DNA: use in lethal lesion determination.

The effect of three radiomodifying agents, cysteamine, hyperthermia, and hypoxia, on the induction of the major classes of X-ray-induced DNA lesions, was studied using mouse L cells and Chinese hamster V79 cells. The use of filter elution techniques allowed most of these studies to be conducted at X-ray doses within the survival-curve range. Cysteamine was found to protect against DNA single-strand breakage (ssb), DNA base damage, and DNA-protein crosslinkage. Hyperthermia had no effect on the level of DNA ssb or DNA base damage, but in L cells (but not in V79 cells) it increased the level of DNA-protein crosslinkage relative to DNA ssb. Hypoxia protected against DNA ssb, had no significant effect on the level of DNA base damage, and enhanced the level of DNA-protein crosslinkage relative to DNA ssb. These results support the previous suggestion that the X-ray-induced lethal lesion is DNA double-strand breakage. Implications of these findings for the mechanisms of formation of X-ray-induced DNA lesions are also discussed.

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PubMed1986

Enhanced induction by X-irradiation of DNA double-strand breakage in mitotic as compared with S-phase V79 cells.

A marked enhancement of X-ray-induced DNA double-strand breakage was demonstrated for mitotic Chinese hamster (V79) cells compared with S-phase cells or asynchronous cultures, which can explain the high radiosensitivity of mitotic cells. However, mitotic cells required twice the level of DNA double-strand breakage to produce a lethal lesion of that required for S-phase or asynchronous cells, suggesting that repair in mitotic cells is less error prone.

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