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Biomedical subjects

J Coppey

Publications and source records attributed to J Coppey.

At least 37 records · Page 2Linked to original sources
PubMed1988

Studies of mutagenesis and neoplastic transformation by bivalent metal ions and ionizing radiation.

We examined the influence of nontoxic concentrations of each of two essential (Zn++ and Mn++) and one nonessential (Ni++) bivalent metal ions on spontaneous and radiation-induced neoplastic transformation and specific gene mutations in mammalian cells. All three metals induced low levels of transformation in mouse BALB/3T3 cells but exerted no mutagenic effect in CHO cells (hprt locus) over a broad range of concentrations. Continuous incubation for 8 or 15 days with each of the metal ions did not enhance the frequency of cell killing, transformation, or mutations induced by acute exposure to x-rays. Zn++, however, had a small but consistent protective effect on the induction of all three endpoints by x-irradiation.

Animals↗
PubMed1987

Promotion of double-strand break repair by human nuclear extracts preferentially involves recombination with intact homologous DNA.

Parameters of DNA double strand break (dsb) repair catalysed by human nuclear extract were analysed using, as substrate, the replicative form (RF) of M13 mp8 in which a single double strand break (dsb) was introduced by restriction. After incubation with the extract, the dsb repair was estimated by the ability of the incubated RF to produce plaques following transfection into JM 109 (Rec A-) bacteria. The possibility of recombination with a purified fragment from M13 mp8 RF enhances up to 20 times the plaquing ability of the RF. The repair by recombination occurs under several conditions: i) the break in the RF must be located in the region of homology with the fragment. ii) the fragment has to be intact in the region corresponding to the break in the RF. iii) a minimal length of homology between the region surrounding the dsb in the RF, and the fragment is required. The in vitro reaction is ATP dependent and dNTP's partially dependent. Dephosphorylation of the free ends in the RF decreases the repair by ligation but is without effect on the recombination.

Cell Nucleus↗
PubMed1987

Homologous recombination intermediates between two duplex DNA catalysed by human cell extracts.

Using as substrates, 1: the replicative form (RF) of phage M13 mp8 in which the reading frame of the lac Z' gene was disrupted by insertion of an octonucleotide, and 2: a restriction fragment one kb long, containing the functional lac Z' gene (isolated from wild type M13 mp8), we show that nuclear extracts from human cells (3 lines tested) promote the targeted replacement of the altered sequence by the functional one. Following incubation with the extracts, the DNA's were introduced in JM 109 bacteria (rec A- and lac Z'-) which were grown in presence of a colorimetric indicator of beta-galactosidase activity. Homologous recombination gives rise to the genotypical modification: lac Z'+ instead of lac Z'- in the bacteriophage DNA. This is revealed by phenotypical expression of the lac Z' gene product in replicating bacteriophage, i.e. the formation of blue instead of white plaques. The frequency of recombination (blue/total plaques) is increased by a factor of 50-80 as a function of protein concentration and of incubation time. The maximal frequency observed is 5 X 10(-5). There is no increase over the background when extracts are boiled. Electrophoresis and electron microscopy of DNA's incubated with the extracts show the formation of recombination intermediates with single strand exchange. Restriction analysis of recombined DNA confirms that the process corresponds to targeted sequence exchange. These data allow to propose three steps for homologous recombination between two duplex DNA's: i) unpairing of the two duplexes; ii) single-strand exchange and synaptic pairing; iii) resolution of the cross-junctions. The three steps correspond to those predicted by the gene conversion model of Holliday.

Cell Line↗
PubMed1986

Toxicity and mutual interactions of cadmium and zinc ions in normal and carcinogen-transformed mouse cells.

The toxicity of chloride salts of physiological (zinc, manganese, nickel) and non-physiological (cadmium) bivalent metal ions was studied in normal or carcinogen-transformed mouse embryo fibroblast cells. The dose response curves for toxicity to both types of cells exhibited similar shapes. The transformed cells, however, were about twice as sensitive to zinc toxicity as normal cells. When normal and transformed cells were grown together and incubated for several hours with an appropriate concentration of zinc, the malignant cells were selectively killed. Cadmium was much more toxic than the three other metal ions in both types of cells. Its toxic effect was reversed by simultaneous addition of zinc at nontoxic concentrations.

Animals↗
PubMed1984

[Lethal effect of the decay of 64Cu and 67Cu incorporated in mammalian cells].

A high lethal efficiency was observed when the decay of two radioactive isotopes of copper, 64Cu or 67Cu, occurred in mammalian cells. The lethal efficiency is of the same order for both isotopes in spite of their different decay processes. As the lethal event could be attributed to an injury inside the cellular DNA only, these results suggest that, even if present in the DNA in trace amounts solely, copper atoms are essential chromatin components. Their behaviour differs depending on whether the cell is a tumor or a non tumor cell because their lethal efficiencies are different (0.5 and 0.1 respectively).

Animals↗
PubMed1983

Lack of reverse transformation after treatment of A 549 human alveolar carcinoma cells with thioproline.

Clonal growth in anchorage dependent (on plastic) and independent (in soft agar) conditions of A 549 cells from a human alveolar carcinoma by thiazolidine-4-carboxylic acid (thioproline) was determined in order to check whether this drug was able to induce reversion towards a normal phenotype of in vivo growth, as described by Gosalvez in HeLa cells. The colony formation on plastic, the ability to form three-dimensional clones in soft agar and cell division in mass culture, were inhibited at similar rates when the cells were grown in the presence of increasing concentrations of thioproline. The morphology of cell clones arising on plastic in the presence of thioproline did not differ from that of untreated cell clones. This indicated that thioproline was unable to provoke 'reverse' transformation of A 549 cells in vitro.

Adenocarcinoma, Bronchiolo-Alveolar↗
PubMed1982

[Application of the lethal effect of electron capture decay: attempt to control the growth of an ascitic krebs tumor in mice using 64cu].

We have recently demonstrated the existence of a lethal effect due to the transmutation of 64Cu atoms associated with the DNA of in vitro cultured cells: normal monkey cells (kidney) and malignant human cells (alveolar lung carcinoma). The lethal efficiency per decay is high and even higher for the malignant cells. From these results, we have tried to bring about a regression of an ascitic tumor developing in the mouse. The experiments we described show that it is possible to clearly delay (5 X 10(5) ascitic cells injected at t = 0), even to stop for a given number of animals (1 to 2 X 10(4) ascitic cells injected at t = 0), the growth of the tumor owing to some injections of 64Cu totalling about 3 mCi and carried out from the 6th day following inoculation of the malignant cells.

Animals↗
PubMed1981

Inhibition of DNA synthesis in relation to enhanced survival of UV-damaged herpes virus in monkey cells treated by a variety of 2-nitronaphthofurans.

Various 2-nitronaphthofuran derivatives (related to each other by simple structural modifications) were tested for 2 different effects in CV-1 monkey kidney cell cultures: the immediate inhibition of normal DNA synthesis and the capacity of pretreated cultures (40 h of contact) to support the replication of UV-damaged Herpes simplex virus (HSV). For all compounds tested, a fair correlation was found between their efficiencies to inhibit cellular DNA synthesis and to provoke an increase in UV-HSV production (virus reactivation). Virus reactivation was due to an increase in both the number of virus-producing cells and the amount of infectious particles produced per cell. The most efficient 2-nitronaphthofurans (particularly 2-nitro-7-methoxy-naphtho[2,1-b]furan-R 7000) were at least as potent as aflatoxin B1 in inducing virus reactivation.

Animals↗
PubMed1981

Enhanced reactivation of ultraviolet-damaged herpes virus in ultraviolet pretreated skin fibroblasts of cancer prone donors.

An enhanced reactivation of ultraviolet-damaged (u.v. at 254 nm) unclear replicating double-stranded DNA viruses occurs when corresponding host cells are treated with radiation or carcinogens prior to infection. This phenomenon seems to be due to an induced DNA repair activity the nature of which is yet unknown. The u.v.-induced enhanced reactivation (ER) of u.v.-damaged herpes simplex virus (u.v. - HSV) was compared in dividing skin fibroblasts of 30 donors either normal or afflicted by genetic disorders, some of which confer a high risk for sunlight induced skin cancers. Cultures were exposed to a single dose of 1.0-25 J.m-2 from 0-60 h before infection with u.v.-HSV (at about 10-3 survival) and the rate of viral production was determined. ER was maximal for a 36 h time interval in all lines. The u.v. dose eliciting maximal ER was 15 J.m-2 in fibroblasts from normal donors, xeroderma pigmentosum (XP) heterozygotes, Mibelli's porokeratosis, diffused naevomatosis, Down's syndrome, xerodermoids, XP variants and epidermodysplasia verruciformis. However, in the latter 3 cases, ER was almost 10 times more pronounced than in the normal cases. The u.v. dose eliciting maximal ER was 0.1, 0.3 and 2 J.m-2 in excision deficient XP fibroblasts from groups A, D and C, respectively, 2.5 J.m-2 in 11961 fibroblasts and 5 J.m-2 in fibroblast lines from cockayne s syndrome.

Adult↗
PubMed1981

Distribution of copper-64 in control mice and in mice bearing ascitic Krebs tumor cells.

Three to 20 hr after an i.p. injection of 64Cu (half-life, 12.8 hr) into mice bearing Krebs ascites cells, a high amount of the radioisotope was recovered in the ascites cells themselves. In the control group, the radioisotope was mainly present in the liver. Similar amounts of 64Cu were recovered in regenerating as well as in normal liver, whereas in the liver of mice bearing ascites cells, this amount was lower by 40 to 50% regardless of the ascitic volume. Thus, the copper metabolism seems to be disturbed at the hepatic level in mice bearing ascites cells. The distribution of 64Cu was 'analyzed in DNA, RNA, and proteins from cellular lysates fractionated by CsCl gradient. There was a uniform pattern of distribution in the macromolecules from ascites cells, while 64Cu' was preferentially associated with the protein fraction from liver. Further experiments indicated that, in vivo, 64Cu was bound to the DNA of ascites cells.

Animals↗
PubMed1980

Survival, DNA synthesis and ribosomal RNA transcription in monkey kidney cells treated by formaldehyde.

Separate cultures of CV-1 cells were exposed for 15 min to 1-16 mM formaldehyde (FA) at various time intervals before labeling with [3H]uridine. The labeled RNA extracted from whole cells was analyzed by polyacrylamide gel electrophoresis. Results indicated that FA produced transcription-terminating lesions in DNA which depressed RNA synthesis, gave rise to shortened RNA chains and modified the expression of transcription-linked genes. These lesions were efficiently repaired since a recovery of RNA transcription, with disappearance of the initially observed alterations, rapidly occurred during post-treatment incubation of cells. Cycloheximide (5 micrograms/ml) strongly inhibited this recovery, whereas fluorodeoxyuridine (10(-5) M) was without effect. The rate of semi-conservative DNA synthesis, measured autoradiographically by thymidine incorporation, as well as the number of cells performing DNA replication, fell to zero after 15-min exposure to FA concentrations greater than 2 mM. Both parameters recovered subnormal levels during a 24-h incubation after treatment with FA concentration up to 8 mM, in agreement with the high survival observed. Unscheduled DNA synthesis was not detectable during the restoration of DNA and RNA synthesis.

Animals↗
PubMed1979

Survival and herpes virus production of normal and xeroderma pigmentosum fibroblasts after treatment with formaldehyde.

The survival of excision-deficient and of excision-proficient (variant) skin fibroblasts from xeroderma pigmentosum (XP) donors was about 5 times and twice, respectively, more sensitive to formaldehyde (FA) treatment than that of skin fibroblasts from healthy and XP heterozygote donors. The capacity of FA-treated host cells to further support Herpes virus (HSV) replication was also more sensitive to FA in XP12BE (group A) than in normal (KD) cells. An important recovery of this capacity occurred in both cell types when they were infected at increasing times (up to 36 h) after FA treatment. This contrasts with the decreasing capacity observed in XP12BE when similarly infected at increasing times after exposure to ultraviolet. In addition, the survival of FA-treated HSV was comparable in KD and XP12BE cells, whereas that of UV-irradiated HSV was much lower in XP12BE than in KD cells.

Cell Line↗
PubMed1979

Herpes virus production as a marker of repair in ultra-violet irradiated human skin cells of different origin.

Confluent cultures of human skin fibroblasts were irradiated with ultra-violet light 0 to 48 hours before infection with herpes simplex virus type 1 (HSV). The one-cycle viral yield was measured. Different responses were obtained according to the origin of the host cells. (1) Cells from three normal donors showed a dose-dependent recovery of HSV production during the 36--40 hours following U.V. exposure. The recovery was maximal for a dose at which a plateau level of unscheduled DNA synthesis (UDS) was reached (24 Jm-2). (2) In a xeroderma pigmentosum (XP) heterozygote line from a mother of XP children, the level of UDS after irradiation up to 48Jm-2 was normal whereas the extent of recovery of HSV production capacity was lower than that of the normal lines. (3) In strains from two cases of XP children, with a normal UDS (XP variants), the recovery process was slowed down and its extent was lower than in normal or XP heterozygote cells. (4) Excision-deficient XP strains from eight cases of XP children presented either no recovery (two strains having the lowest UDS, less than 2 per cent) or a small recovery, the extent of which was in good agreement with the corresponding level of UDS (between 5 and 30 per cent). Measurement of this recovery seems to be a very sensitive assay for detecting differences in the repair abilities of U.V.-irradiated human skin cells of various origins.

Cells, Cultured↗
PubMed1978

Herpes virus production as a marker of repair in ultraviolet irradiated human skin cells of different origin.

When confluent human skin cultures are ultraviolet (UV)-irradiated before infection with Herpes Simplex type 1 virus (HSV), their capacity to support virus growth is impaired. When the time interval between UV-exposure and infection is increased up to 36 hours, different recoveries of HSV production capacity are observed according to the origin of the host cells. 1) Two normal donors: the cells present a dose dependent recovery which is maximal for a dose ( : formula: (see text) at which a plateau level of unscheduled DNA synthesis (UDS) is reached. 2) A mother of two Xeroderma Pigmentosum (XP) children: in this line which exhibits a normal level of UDS, the extent of recovery is significantly decreased after exposures : formula: (see text) 3) An XP child: these cells have a normal level of UDS (XP variant) whereas they present a low extent of recovery as compared with that of the normal subjects. 4) Five XP children: in these excision deficient lines (UDS less than 15%), HSV production capacity decreases with increasing time intervals after UV exposure for doses greater than or equal to 3 : formula: (see text). For doses less than 3 : formula: (see text), a small recovery with an overshoot of viral production is observed 24 h after UV exposure in the lines (three) which present the highest UDS (10--15%) and not in the two lines which present a very low UDS (1--2%).

Adult↗