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1.
As was reported elsewhere (Gasparian, Grigorian, 1989a, 1989b), the stimulation of cell proliferation takes place in established culture of chick embryo cells after adding a suspension of living or inactivated homologous cells. In the present paper the kinetic parameters of this process, termed as the contact stimulation of cell proliferation, were studied. The dose- and time-dependence of cell response to the stimulus is described. It was shown that the addition of cells activates cell growth both in exponential and stationary cultures. DNA synthesis in resting cells is seen initiated only if their continuous interaction with the added cells is provided. The nature of signals involved in the process of contact stimulation is described. 相似文献
2.
The secondary culture of chick embryo cells in 2-3 days after seeding was super-inoculated with homologous cells. Suspended cells adhere and spread on the cell layer whereby the culture density increases quickly. After adhesion of exogenous cells to the layer the stimulation of cell proliferation takes place. This activation is not connected with methodical manipulations or with the influence of conditioned medium factors. The results suggest that the increase in cell number itself does not arrest cell multiplication. It is proposed that the known phenomenon of blocking cell proliferation in dense cultures cannot be attributed only to effects of high cell density. 相似文献
3.
The secondary cultures of chick embryo cells were suspended and transferred to homologous cell cultures. Cell adhesion and proliferation were studied in these superinoculated cultures. It was shown that added cells soon adhered to the underlying cell layer which results in a prompt increase in culture density followed by the activation of DNA synthesis and cell division. Stimulation of cell proliferation involved both cell subpopulations composing the superinoculated culture: cells seeded on the built-up cell layer and cells of the layer. The contact nature of added cell mitogenic action on overlaid cell proliferation was evidenced. The cell system described can be used to investigate the adhesive properties of the cell layer apical surface, the relationship between cell growth rate and culture density, and the contact stimulation of cell proliferation. 相似文献
4.
Altered baby hamster kidney (BHK-R) cells were serially cultured in the continuous presence of hemagglutinating virus of Japan (HVJ). These cells showed a distinct resistance to superinfection with the homologous HVJ. This resistance of BHK-R cells gradually disappeared after serial passages in the presence of ultraviolet-irradiated HVJ particles which lost infectivity but still preserved hemagglutinating and neuraminidase activities. When BHK-R cells were serially cultured in the presence of a temperature-sensitive mutant of HVJ at non-permissive temperature, the cells also lost the resistance. The resistance of BHK-R cells remained unchanged, even after prolonged incubation in virus-free maintenance medium under the conditions of no cell division. It was suggested that killing of virus-sensitive cells, which were generated during cell proliferation, was required for maintenance of the resistance. 相似文献
5.
cDNA cloning and sequence analysis of beta ig-h3, a novel gene induced in a human adenocarcinoma cell line after treatment with transforming growth factor-beta. 总被引:18,自引:0,他引:18
J Skonier M Neubauer L Madisen K Bennett G D Plowman A F Purchio 《DNA and cell biology》1992,11(7):511-522
Transforming growth factor-beta (TGF-beta) is capable of affecting the proliferation of many cell types. To identify novel genes whose protein products may mediate cellular responses to this factor, a cDNA library was made from mRNA isolated from a human lung adenocarcinoma cell line (A549) that had been treated for 3 days with TGF-beta. The library was screened by differential hybridization and a cDNA clone, beta ig-h3, was isolated. This gene was induced up to 20-fold in A549 cells after 2 days of treatment with TGF-beta 1. It was also induced in several other cell lines, including PC-3 and H2981. DNA sequence analysis of beta ig-h3 indicated that it encoded a novel protein, beta IG-H3, of 683 amino acids, which contained an amino-terminal secretory sequence and a carboxy-terminal Arg-Gly-Asp (RGD) sequence that can serve as a ligand recognition site for several integrins. beta IG-H3 also contained short amino acid regions homologous to similar regions in Drosophila fasciclin-I and four homologous internal domains, which can be folded into a potential bivalent structure and could act as a bridge between cells expressing the appropriate ligand. beta ig-h3 RNA was detected in several cell lines and tissues. COS cells transfected with plasmids encoding beta IG-H3 secreted a major 68-kD protein that was detected by immunoblotting using antipeptide antibodies. Since beta ig-h3 is induced in several cell lines whose proliferation is affected by TGF-beta 1, it may be involved in mediating some of the signals of this multifunctional growth modulator. 相似文献
6.
Regulation of double-strand break-induced mammalian homologous recombination by UBL1, a RAD51-interacting protein 总被引:6,自引:2,他引:4
Li W Hesabi B Babbo A Pacione C Liu J Chen DJ Nickoloff JA Shen Z 《Nucleic acids research》2000,28(5):1145-1153
Mammalian RAD51 protein plays essential roles in DNA homologous recombination, DNA repair and cell proliferation. RAD51 activities are regulated by its associated proteins. It was previously reported that a ubiquitin-like protein, UBL1, associates with RAD51 in the yeast two-hybrid system. One function of UBL1 is to covalently conjugate with target proteins and thus modify their function. In the present study we found that non-conjugated UBL1 forms a complex with RAD51 and RAD52 proteins in human cells. Overexpression of UBL1 down-regulates DNA double-strand break-induced homologous recombination in CHO cells and reduces cellular resistance to ionizing radiation in HT1080 cells. With or without overexpressed UBL1, most homologous recombination products arise by gene conversion. However, overexpression of UBL1 reduces the fraction of bidirectional gene conversion tracts. Overexpression of a mutant UBL1 that is incapable of being conjugated retains the ability to inhibit homologous recombination. These results suggest a regulatory role for UBL1 in homologous recombination. 相似文献
7.
Miyano-Kurosaki N Kira J Barnor JS Maeda N Misawa N Kawano Y Tanaka Y Yamamoto N Koyanagi Y 《Microbiology and immunology》2007,51(2):235-242
That HTLV-I infects CD4(+) T cells and enhances their cell growth has been shown as successful long-term in vitro proliferation in the presence of IL-2. It is known that T cells isolated from HAM patients possess strong ability for cell proliferation in vitro and mRNA of various cytokines are abundantly expressed in CNS tissues of HAM patients. Hence, the cytokine-induced proliferation could have an important role in pathogenesis and immune responses of HAM. In this study, we examined the relationship between cell proliferation and ability of in vitro cytokine production of CD4(+) T cell clones isolated from HAM patients. We started a culture from a single cell to isolate cell clones immediately after drawing blood from the patients using limiting dilution method, which could allow the cell to avoid in vitro HTLV-I infection after initiation of culture. Many cell clones were obtained and the rate of proliferation efficiency from a single cell was as high as 80%, especially in the 4 weeks' culture cells from HAM patients. These cells were classified as mainly Th0 phenotype that produce both IFN-gamma and IL-4 after CD3-stimulation. However, the frequency of proviral DNA in these cloned cells was significantly low. Our results indicate that the ability of cell proliferation in HAM patients is not restricted in HTLV-I-infected T cells. HTLV-Iuninfected CD4(+) T cells, mainly Th0 cells, also have a strong ability to respond to IL-2-stimulation, showing that unusual immune activation on T cells has been observed in HAM patients. 相似文献
8.
H Y Zhou Y Ohnuma H Takita R Fujisawa M Mizuno Y Kuboki 《Biochemical and biophysical research communications》1992,186(3):1288-1293
A lysine-rich 18 kDa protein was isolated from bovine bone and examined for its effects on osteoblast-like MC3T3-E1 cells. This protein is homologous to a heparin-binding protein in brain and uterus. This protein enhanced cell attachment independent of the Arg-Gly-Asp cell-binding sequence and stimulated proliferation during the growth phase. Addition of this protein to cell cultures on days 11, 12, and 13 after confluency resulted in a 1.6-2.0-fold increase in the alkaline phosphatase activity and little increase in the DNA content. These findings suggest that the 18 kDa protein may be functional in promoting the proliferation and differentiation of osteoblasts. 相似文献
9.
R. P. C. Rijke R. Gart 《Virchows Archiv. B, Cell pathology including molecular pathology》1979,31(1):15-22
Epithelial cell loss was induced in the descending colon of the rat by temporary ischaemia to investigate whether this would lead to an increase in crypt cell proliferation. Shortly after the temporary ischaemia the number of cells per crypt was markedly reduced, and it was shown that the cell loss occurred mainly from the non-proliferating upper half of the crypt. The number of cells per crypt reached control values again after 24-48 h. There was a marked increase in proliferative activity, as reflected by the labelling index after 3HTdR and by the mitotic index, with peak values at 16 and 24 h after ischaemia. After 48 h the proliferative indices were normal again. The increase in crypt cell proliferation was characterized by an increase in the labelling index as well as in the mitotic index per crypt cell position. No enlargement of the proliferative cell compartment in the crypt was observed. It is most likely then that the increase in crypt cell proliferation was brought about by a shortening of the cell cycle, since the growth fraction in the lower half of the crypt approaches 1.0. The possible implications of the present data for the control of colonic cell proliferation and colonic carcinogenesis are discussed. 相似文献
10.
Neurogenesis in the crustacean ventral nerve cord: homology of neuronal stem cells in Malacostraca and Branchiopoda? 总被引:4,自引:0,他引:4
Steffen Harzsch 《Evolution & development》2001,3(3):154-169
SUMMARY In Insecta and malacostracan Crustacea, neurons in the ventral ganglia are generated by the unequal division of neuronal stem cells, the neuroblasts (Nbs), which are arranged in a stereotyped, grid‐like pattern. In malacostracans, however, Nbs originate from ectoteloblasts by an invariant lineage, whereas Nbs in insects differentiate without a defined lineage by cell‐to‐cell interactions within the neuroectoderm. As the ventral ganglia in entomostracan crustaceans were thought to be generated by a general inward proliferation of ectodermal cells, the question arose as to whether neuroblasts in Euarthropoda represent a homologous type of stem cell. In the current project, neurogenesis in metanauplii of the entomostracan crustaceans Triops cancriformis Fabricius, 1780 (Branchiopoda, Phyllopoda) and Artemia salina Linné, 1758 (Branchiopoda, Anostraca) was examined by in vivo incorporation of the mitosis marker bromodeoxyuridine (BrdU) and compared to stem cell proliferation in embryos of the malacostracan Palaemonetes argentinus Nobili, 1901 (Eucarida, Decapoda). The developmental expression of synaptic proteins (synapsins) was studied immunohistochemically. Results indicate that in the ventral neurogenic zone of Branchiopoda, neuronal stem cells with cellular characteristics of malacostracan neuroblasts are present. However, a pattern similar to the lineage‐dependent, grid‐like arrangement of the malacostracan neuroblasts was not found. Therefore, the homology of entomostracan and malacostracan neuronal stem cells remains uncertain. It is now well established that during arthropod development, identical and most likely homologous structures can emerge, although the initiating steps or the mode of generation of these structures are different. Recent evidence suggests that adult Entomostraca and Malacostraca share corresponding sets of neurons so that the present report provides an example that those homologous neurons may be generated via divergent developmental pathways. In this perspective, it remains difficult at this point to discuss the question of common patterns of stem cell proliferation with regard to the phylogeny and evolution of Atelocerata and Crustacea. 相似文献
11.
12.
N. V. Panyukhin K. S. Vishnyakova Y. E. Yegorov 《Biochemistry (Moscow) Supplemental Series A: Membrane and Cell Biology》2008,2(4):326-332
Effects of partial oxygen pressure in a gas mixture surrounding the culture medium on survival, proliferation and differentiation of mesenchymal stem cells (MSC) isolated from mouse bone marrow was studied. It was found that 3% oxygen increased the survival of cells seeded at low density; the rate and duration of the MSC proliferation were also elevated. Effect of oxygen concentration on replicative activity of MSC was manifested as early as the first few days after the onset of cell growth. Studies of colony formation revealed that preincubation of cells with 21% oxygen had a negative effect on cell growth under 3% oxygen, while preincubation with 3% oxygen stimulated MSC proliferation in the presence of 21% oxygen. Notwithstanding, this effect of oxygen cannot be interpreted as unequivocally deleterious. Low partial oxygen pressure inhibited osteogenic differentiation of MSC, but adipogenic differentiation was insensitive to oxygen concentration. It is concluded that proliferation and differentiation of MSC depend critically on oxygen content in the culture medium. 相似文献
13.
Gilles A. Spoden Ursula Rostek Stefan Lechner Maria Mitterberger Sybille Mazurek Werner Zwerschke 《Experimental cell research》2009,315(16):2765-2774
The glycolytic key regulator pyruvate kinase M2 (M2-PK or PKM2) can switch between a highly active tetrameric and an inactive dimeric form. The transition between the two conformations regulates the glycolytic flux in tumor cells. We developed specific M2-PK-binding peptide aptamers which inhibit M2-PK, but not the 96% homologous M1-PK isoenzyme. In this study we demonstrate that, at normal blood glucose concentrations, peptide aptamer-mediated inhibition of M2-PK induces a significant decrease of the population doubling (PDL rate) and cell proliferation rate as well as an increase in cell size, whereas under glucose restriction an increase in PDL and cell proliferation rates but a decrease in cell size was observed. Moreover, M2-PK inhibition rescues cells from glucose starvation-induced apoptotic cell death by increasing the metabolic activity. These findings suggest that M2-PK is a metabolic sensor which regulates cell proliferation, cell growth and apoptotic cell death in a glucose supply-dependent manner. 相似文献
14.
Calcium response after stimulation by substance P of U373 MG cells: inhibition of store-operated calcium entry by protein kinase C 总被引:2,自引:0,他引:2
In this paper we investigate the Ca2+ response after Substance P (SP) stimulation of U373 MG cells. SP is a tachykinin and physiologically acts as a neurotransmitter and neuromodulator in the nervous system, but pathologically triggers malignant glial cells, such as U373 MG, to release cytokines and increase proliferation rate.In this paper we show that SP increases the proliferation rate of U373 MG cells and the intracellular Ca2+ concentration by mobilizing Ca2+ only from thapsigargin-sensitive stores. In fact, Ca2+ entry through store-operated calcium entry (SOCE) channels, which was observed after thapsigargin treatment, was not detected after stimulation by SP. The inhibition of SOCE after SP stimulation must be mediated by protein kinase C (PKC), because it was not observed in the presence of calphostin C (an inhibitor of PKC). Moreover, stimulation by SP-induced membrane potential hyperpolarization. Our results are consistent with the following sequence of events: (i) SP interacts with NK(1) receptors; (ii) fast homologous receptor desensitization occurs; (iii) reuptake by endoplasmic reticulum Ca(2+)-ATPase quantitatively overwhelms the extrusion by plasma membrane Ca2+-ATPase. These results have two important consequences. In U373 MG cells the SOCE does not contribute to the Ca2+ response after SP, and is not necessarily involved in promoting cell proliferation. 相似文献
15.
Endonuclease G (EndoG) is a mitochondrial apoptosis regulator that also has roles outside of programmed cell death. It has been implicated as a defence DNase involved in the degradation of exogenous DNA after transfection of mammalian cells and in homologous recombination of viral and endogenous DNA. In this study, we looked at the effect of EndoG depletion on plasmid DNA uptake and the levels of homologous recombination in HeLa cells. We show that the proposed defence role of EndoG against uptake of non-viral DNA vectors does not extend to the cervical carcinoma HeLa cells, as targeting of EndoG expression by RNA interference failed to increase intracellular plasmid DNA levels. However, reducing EndoG levels in HeLa cells resulted in a statistically significant reduction of homologous recombination between two plasmid DNA substrates. These findings suggest that non-viral DNA vectors are also substrates for EndoG in its role in homologous recombination. 相似文献
16.
Hypothalamic Neurons in Dissociated Cell Culture: The Mechanism of Increased Survival Times in the Presence of Non-Neuronal Cells 总被引:3,自引:2,他引:1
Rat hypothalamic neurons were cultured in the presence of fluorodeoxyuridine to inhibit nonneuronal cell proliferation. Under these conditions, neuronal cell survival was dependent on contact with homologous nonneuronal cells. This phenomenon did not seem to be due to the release of diffusable trophic factors, since neither growth on polylysine in the close proximity of nonneuronal cells nor the use of preconditioned medium significantly increased neuronal survival. However, metabolically active cell layers were required, since growth on heat-killed or fixed homologous nonneuronal cells did not increase neuronal survival. The increased survival of neurons seen here in the presence of homologous nonneuronal cells therefore appears to be due to metabolic co-operation mediated by specific, direct cell-cell contact. 相似文献
17.
20 KDa homologous restriction factor of complement resembles T cell activating protein 总被引:17,自引:0,他引:17
H Okada Y Nagami K Takahashi N Okada T Hideshima H Takizawa J Kondo 《Biochemical and biophysical research communications》1989,162(3):1553-1559
We previously identified a 20KDa membrane glycoprotein 1F5 antigen which inhibits the assembly of homologous complement membrane attack complexes and we designate it as HRF20 standing for 20KDa homologous restriction factor. The amino acid sequence deduced from its coding base sequence resembles that of T cell activating protein, most conspicuously in cysteine residues, 10 out of 11 of which occupy identical positions in an overall sequence homology of 24.8%. Furthermore, proliferation of human T cells was stimulated by monoclonal antibody to HRF20. 相似文献
18.
DNA repair and cell survival in haploid and its diploid derivative strains ofSaccharomyces cerevisiae were studied after 100 krad X-ray irradiation. The cells were in theG
1 stage of the cell cycle, where haploid cells had only one copy of genetic material per genome and diploid had two copies. It was found that diploid could repair double-strand breaks in its DNA after 48 hr of liquid holding which was accompanied by a four-fold rise in survival. In contrast a haploid strain failed to repair its DNA and showed no increase in survival after liquid holding. It is concluded that (1) repair of DNA double-strand breaks requires the availability of two homologous DNA duplexes, (2) restoration of cell viability during liquid holding is connected with repair of DNA double-strand breaks and (3) this repair is a slow process possibly associated with slow finding and conjugation of homologous chromosomes. 相似文献
19.
The percentage of labeled cells in the uterine luminal epithelium of cycling mice showed the different zonal distributions at each stage of estrous cycle after cumulative labeling with 3H-thymidine for 36 hr. It was estimated that the proliferating fraction in the epithelium at proestrus, estrus, metestrus, and diestrus was 100%, 100%, 40% and 5%, respectively. The percentage of labeled cells in the uterine luminal epithelium of cycling mice treated with progesterone remained below 10% level for at least 20 hr after injections of progesterone. Total labeling was attained in the uterine epithelium of castrated mice by the administration of estradiol-17beta. On the other hand, the cell proliferation in the uterine epithelium of castrated mice treated with estradiol and progesterone was markedly suppressed and the percentage of labeled cells remained approximately at 35%. The remaining cell population, however, still showed the mitotic potency when mice received estradiol. It is suggested from this study that the effect of progesterone is to suppress the epithelial cell proliferation and transfer cells into resting cell fraction which is still evoked to proliferate as the effect of estradiol and that a key factor controlling epithelial proliferation in mouse uterus during the estrous cycle is proliferating fraction rather than cell cycle time. 相似文献
20.
H R MacDonald 《Cellular immunology》1978,35(2):414-426
Some physical and proliferative characteristics of cytolytic thymus-derived lymphocytes (CTL) have been investigated in long-term mixed leukocyte cultures (MLC). Velocity sedimentation analysis of MLC cells restimulated by homologous alloantigens at low responding cell density indicated that a shift from large cycling CTL to much smaller (probably non-cycling) CTL occurred between the third and sixth day in secondary cultures. This change in physical characteristics as a function of growth phase was accompanied by a parallel change in the responsiveness of secondary MLC cells to a further alloantigenic stimulus; restimulated Day 3 secondary cells gave rise to a transient CTL response (peaking after 2–3 days) whereas the response of restimulated Day 6 secondary cells increased for 4 days and reached much higher peak levels. Repeated stimulation of MLC cells under the latter conditions led to dramatic increases in both CTL activity and viable cell number. In particular, four sequential restimulations at 7-day intervals resulted in a calculated absolute increase of approximately 500,000-fold in both parameters. Cells derived from such extensive proliferation retained their original lytic specificity and were uniquely T cells as determined by surface markers. These results raise interesting questions regarding the extent and regulation of CTL proliferation in MLC. 相似文献