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N A Setkov

Publications and source records attributed to N A Setkov.

At least 19 recordsLinked to original sources
PubMed2004

[Different sensitivity of inbred mice to hepatocarcinogen ortho-aminoazotoluene may be due to differences in the negative control mechanisms of hepatocyte proliferation].

A most convenient model to study mechanisms of live organism response to chemical carcinogens is tumor induction in murine liver by aminoazodyes, in particular by ortho-aminoazotoluene (OAT). We studied both early and late stages of hepatocarcinogenesis on several lines of inbred mice differing in sensibility to OAT. By means of autoradiography, we examined proliferative activity of hepatocytes obtained from the liver of sensitive (A/He, DD, SWR) and resistant to OAT AKR, CC57Br, BALB/c lines of mice, which were injected carcinogen. The level of p53, p21Cip1, bax, mdm2, cyclin G, gadd45 genes expression in the liver of mice of different lines given OAT injection was studied by multiplex PCR method. Carcinogen caused a decrease of hepatocyte proliferative activity induced by partial hypatectomy (PHE), and an increase in p53, p21Cip, bax, mdm2, and cyclin G genes within mice of A/He, DD and SWR lines. Cell fusion experiments on hepatocytes obtained from regenerating murine liver sensitive to A/He line carcinogen and given long-time OAT administrations with resting and proliferating fibroblasts of NIH 3T3 mice revealed no obvious suppression of DNA synthesis in heterokaryons. Unlike, in fusion experiments on serum-stimulated fibroblasts with hepatocytes obtained from the liver of BALB/c line mice also given OAT suppression of DNA synthesis in stimulated fibroblasts in heterokaryons was observed 15 days following PHE. These results enable us to conclude that OAT administrations break negative endogenous mechanisms of hepatocyte proliferation control in the liver of mice sensitive to carcinogenes.

3T3 Cells↗
PubMed2002

Dobutamine prevents experimental postintoxication liver cirrhosis in mice.

Various chronic inflammatory and necrotic processes in the liver parenchyma are accompanied by pathological morphofunctional changes, which are associated with hepatocyte death and hyperplasia of the connective tissue. Regeneration of the liver parenchyma should include not only prevention of fibrosis, but also stimulation of hepatocyte proliferation. The adrenoceptor agonist dobutamine stimulated proliferative activity of cultured hepatocytes and prevented the development of postintoxication liver cirrhosis in mice produced by chronic poisoning with CCl(4).

Adrenergic beta-Agonists↗
PubMed2001

[Hepatocytes in heterokaryons can suppress entry into the S-period of fibroblast nuclei from murine NIH 3T3 cells].

Mouse liver regeneration after partial hepatectomy can be considered as a spectacular example of controlled tissue increase. In this study serum-deprived (0.2%) resting and serum-stimulated (10%) proliferating NIH 3T3 mouse fibroblasts were fused with primary hepatocytes isolated from normal (intact) and regenerating adult mouse liver at different times after partial hepatectomy (1-15 days) to elucidate mechanisms of liver cell proliferation cessation at the regeneration end. DNA synthesis was investigated in the nuclei of heterokaryons and non-fused cells using radioautography. Hepatocytes isolated from regenerating liver within 1-12 days following operation did not retard the entry of stimulated fibroblast nuclei into the S-period. In contrast, hepatocytes isolated within 15 days after hepatectomy were found to have inhibitory effect on the entry of stimulated fibroblast nuclei into the S-period in heterokaryons. Preincubation of these hepatocytes with cyclocheximide for 2-4 h abolished their ability to suppress DNA synthesis in stimulated fibroblast nuclei in heterokaryons. Possible reasons of inhibitory effect of differentiated cells in heterokaryos are discussed. The data obtained enable us to conclude that the mechanism of proliferative process control in regenerating hepatocytes seems to be stopped being affected by the intracellular growth inhibitors, whose formation depends on protein synthesis.

3T3 Cells↗
PubMed1997

[The effect of intracellular protease inhibitors on DNA synthesis in fibroblasts from the NIH 3T3 mouse strain].

The enhancement of catabolic reactions of cell macromolecules, primarily proteins, raises the question about the role of active proteolysis and particular kinds of proteases in the process leading to cell proliferation retardation. The application of highly specific inhibitors of intracellular proteases helps to answer this question even in part. It has been shown that a short-term (2-6 h) treatment of resting cells of line NIH 3T3 with inhibitors of lysosomal cysteinous proteases--cystatine or leupeptin, as well as with inhibitors of non-lysosomal Ca(2+)-activated cystein proteases--L-cystamine and L-cystin--enhances the cell proliferative response on the serum and even stimulates tritiated thymidine incorporation in the cells cultured in the medium with a low (0.2%) serum content. Unlike, the treatment of resting cells with the lysosomotropic agent chloroquin, known to shut down all lysosomal hydrolases, reduces the cell proliferative response of the serum. In the experiments of fusing the resting and stimulated cells, it was found that the pretreatment of the former with inhibitors of lysosomal proteases enhanced the index of labeled nuclei in the latter, while observed in dikaryons, compared to the control, although failed to remove totally the effect of DNA synthesis inhibition in heterokaryons (Setkov et al., 1984; Setkov, Kazakov, 1989). A conclusion is made that the short-term metabolic shift in resting cells towards anabolism may result in that resting cells commit a cycle, and that an enhanced activity of cystein proteases is one of specific metabolic processes in resting cells leading to proliferation retardation.

3T3 Cells↗
PubMed1997

Brief exposures of resting fibroblasts to okadaic acid stimulate DNA synthesis.

To study further the factors providing for cellular quiescence, we used okadaic acid (OA) at concentrations (0.1, 1, 10 or 100 nM) inhibiting type 1 and/or type 2A protein phosphatases in mammalian cell cultures. Brief (2 h) exposure of resting (0.2% serum for 72 h) NIH 3T3 mouse fibroblasts to OA with subsequent incubation of cells in a medium with 0.2% serum, stimulated DNA synthesis at all concentrations studied. Maximal stimulation was observed following pre-incubation of resting cells with 10 nM OA. Treatment of cycling cells (10% serum) with OA (2 h pulses at 12 h intervals for 72 h) prevented their exit to the resting state on transfer to a medium with 0.2% serum. Brief exposures of resting cells to OA did not affect the rate of protein synthesis. OA pulses in the late pre-replicative period had no effect on the entry of serum-stimulated cells into the S phase. Cell fusion experiments with resting (serum-deprived) and proliferating (serum-stimulated) NIH 3T3 cells, using radioautography with a double-labelling technique, revealed that pre-incubation of resting cells with OA for 2 h before and after fusion abrogates their ability to suppress the onset of DNA synthesis in the nuclei of proliferating cells in heterodikaryons. The results indicate that protein phosphatases of type 1 and/or 2A may be involved in the growth-arrest machinery that provides for cellular quiescence.

3T3 Cells↗
PubMed1996

[DNA synthesis in heterodikaryons obtained by the fusion of long-term stimulated cells of the NIH 3T3 line with short-term stimulated cells].

Serum-deprived (0.2%) resting NIH 3T3 mouse fibroblasts, stimulated for 0.5-4.0 h by serum (10%) or PDGF (beta-beta homodimer) (early cells), were fused with stimulated cells taken 10 h after changing the medium to one containing 10% serum DNA synthesis was investigated in the nuclei of monokaryons, homodikaryons and heterodikaryons, using radioautography with the double-labeling technique. Preincubation of resting cells with serum for 3 h or with the competence factor PDGF for 1.0-1.5 h abolished their ability to suppress DNA synthesis in late stimulated nuclei in heterodikaryons. Actomycin D restores the inhibitory capacity of resting cells stimulated by PDGF but not by serum. The results give evidence that the acquirement by resting cells of competence for DNA replication includes as a necessary step the down-regulation of intracellular growth inhibitors, and thus serum can overcome the influence of endogenous growth inhibitors, at any rate, by two ways.

3T3 Cells↗
PubMed1995

Transient inhibition of protein synthesis induces expression of proto-oncogenes and stimulates resting cells to enter the cell cycle.

There is evidence that resting cells are able to produce molecules with antiproliferative activity, some of which behave as short-lived repressor proteins. We suggest that transient inhibition of protein synthesis in resting cells would lead to a decrease in the levels of these negative growth regulators and might, therefore, promote mitogenic responses. We report that treatment of resting (serum-deprived) NIH 3T3 cells with cyclocheximide (CH) or puromycin induces expression of c-fos, c-jun and c-myc proto-oncogenes in a manner similar to that of platelet-derived growth factor (PDGF). Actinomycin D (Act D) abrogates the induction of proto-oncogene expression. Transient inhibition of protein synthesis by CH or puromycin also induces the resting NIH 3T3 and C3H 1OT1/2 cells to enter the cell cycle. Inhibition of new RNA or protein synthesis abolishes the proliferative response. These findings show that control mechanisms at both transcriptional and translational levels are operative in the resting cells treated with protein synthesis inhibitors. Cell fusion experiments with resting and serum-stimulated NIH 3T3 cells revealed that brief pre-incubation of resting cells with either PDGF, CH or puromycin abrogates their ability to suppress the onset of DNA synthesis in the nuclei of stimulated cells in heterodikaryons. However, the abrogative effect of PDGF disappeared in the presence of Act D, whereas the effects of protein synthesis inhibitors did not, indicating their independence of the induction of transcription. The data suggest that the observed effects of protein synthesis inhibitors are connected with elimination of some short-lived negative growth regulators, since a brief translational arrest is sufficient for the resumption of DNA synthesis in the nuclei of stimulated cells blocked by resting cells in heterodikaryons.

3T3 Cells↗
PubMed1992

Protein synthesis inhibitors, like growth factors, may render resting 3T3 cells competent for DNA synthesis: a radioautographic and cell fusion study.

Serum-deprived (0.1-0.2%) resting NIH 3T3 mouse fibroblasts pre-incubated with cycloheximide (7.5 micrograms/ml), or puromycin (10 micrograms/ml), were fused with stimulated cells taken 10 h after changing the medium to one containing 10% serum, and DNA synthesis was investigated in the nuclei of monokaryons, homodikaryons and heterodikaryons using radioautography with the double-labelling technique. Pre-incubation of resting cells with inhibitors of protein synthesis for 1-4 h abolished their ability to suppress DNA synthesis in stimulated nuclei in heterokaryons. Three hours after the removal of cycloheximide from the medium, the resting cells acquired once again the inhibitory capacity for entry of stimulated nuclei into the S period. This inhibitory influence disappeared also in the case of post-fusion cycloheximide application as well as following an 8-12 h pre-treatment of resting cells with actinomycin D (1 microgram/ml) prior to fusion. Pre-incubation of resting cells for 12 h with PDGF (1 u/ml-1) followed by an 8-48 h incubation in serum-free medium stimulated the onset of DNA synthesis. A brief exposure (45 min) of resting cells to cycloheximide (7.5 micrograms/ml), or puromycin (7.5 micrograms/ml), exerted a similar effect, inducing by itself the entry of cells into the S period. The results support the assumption that acquirement, by resting cells, of competence for DNA replication includes as a necessary step the down-regulation of intracellular growth inhibitors whose formation depends on protein synthesis.

3T3 Cells↗
PubMed1991

[Hepatocytes from the liver of mice with experimental post-toxic cirrhosis stimulate in heterokaryons DNA synthesis in the nuclei of resting fibroblasts of the NIH 373 line].

Hepatocytes from mouse liver with experimental post-toxic cirrhosis (received by means of 10-12 inhalations with CCl4) were fused with serum-deprived (0.2%) resting NIH 3T3 mouse fibroblasts to elucidate mechanisms of liver stroma cells proliferation at cirrhosis. After fusion, nuclei of fibroblasts in such heterokaryons were found to enter into S-period without any exogenous stimulation of cell proliferation (in the medium with low content of serum). The obtained data allow us to suggest that hepatocytes from mouse liver with experimental post-toxic cirrhosis can produce and secrete into the medium (blood) factor (s) capable of stimulating the mesenchymal origin cell proliferation.

Animals↗
PubMed1991

[Hepatocytes in heterokaryons do not retard the nuclei of stimulated fibroblasts entering the S period].

Serum-deprived (0.2%) resting and serum-stimulated (10%) proliferating NIH 3T3 mouse fibroblasts were fused with hepatocytes from intact, regenerating and embryonic mouse livers to elucidate mechanisms of liver cell proliferation, DNA synthesis being investigated in nuclei of heterokaryons and non-fused cells using radioautography. Hepatocytes in heterokaryons were found to have no inhibitory effect on the entry of stimulated fibroblast nuclei into the S-period, but on the contrary they were involved in DNA synthesis. In addition, the nuclei in heterokaryons mutually stimulated each other to enter the S-period. In their turn, the resting fibroblasts did not prevent the proliferating hepatocytes from the regenerating and embryonic livers to enter the S-period. Possible reasons of the absence of inhibitory effect of differentiated cells in heterokaryons are discussed. The data obtained enable us to conclude that the mechanism of proliferative process control in resting immortalized cells differs from that in differentiated cells where proliferation seems to be stopped without affecting the endogenous inhibitor postulated for the resting and ageing fibroblasts.

3T3 Cells↗
PubMed1991

[The effect of cycloheximide on the entry into the S period of the nuclei in heterodikaryons].

Serum-deprived (0.2%) resting NIH 3T3 mouse fibroblasts were fused with serum-stimulated (10%) proliferating cells to elucidate mechanisms of entering into S-period operating in the nuclei of the heterokaryons under the effect of cycloheximide--an inhibitor of protein synthesis. Using radioautography DNA synthesis was investigated in mono-, homo- and heterodikaryons. After short (0.5-3.0 h) depressing of protein synthesis, the nuclei of stimulated cells in heterokaryons were found to enter into S-period. Under these conditions no induction of DNA synthesis was found in the nuclei of resting cells in heterodikaryons. In other experiments, resting cells were under the effect of cycloheximide during 2-4 h before the fusion, that led to a great induction of DNA synthesis in the nuclei of these cells in heterodikaryons. The data obtained are consistent with the idea of fibroblast transition to the rest under the action of labile proteins-repressors.

3T3 Cells↗