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1.
Inhibition of proliferation of cultured rat liver epithelial cells at specific cell cycle stages by transforming growth factor-beta 总被引:1,自引:0,他引:1
P Lin C Liu M S Tsao J W Grisham 《Biochemical and biophysical research communications》1987,143(1):26-30
Proliferation of early-passage propagable cultured rat liver epithelial cells derived from normal adult rats is markedly inhibited by transforming growth factor-beta (TGF-beta). Inhibition, which is completely reversible, is effected at two distinct points of the cell cycle, the G1/S border and the G0 or early G1 phase. With increasing passages in culture, hepatic epithelial cells progressively become less sensitive to the inhibitory effect of TGF-beta. 相似文献
2.
E J van Zoelen T M van Oostwaard P T van der Saag S W de Laat 《Journal of cellular physiology》1985,123(2):151-160
Polypeptide growth factor activity in serum can be destroyed by treatment with dithiothreitol. When such growth-factor-inactivated serum is used as a supplement of culture media instead of regular serum, normal rat kidney (NRK) cells become quiescent unless defined polypeptide growth factors like insulin and epidermal growth factor (EGF) are added. On this basis a growth-factor-defined medium has been developed for NRK cells, which permits cell proliferation as rapidly as in media supplemented with serum, even at low cell densities. Moreover, cells can be serially passaged in this medium. NRK cells can be induced to grow in semisolid media when incubated with transforming growth factors. The growth-factor-defined medium permits soft agar growth experiments of NRK cells, without interference from polypeptide growth factors in serum. Using this assay system we have shown that EGF alone is unable to induce any degree of anchorage-independent growth in NRK cells. However, a recently identified transforming growth factor from mouse neuroblastoma cells which does not compete with EGF for receptor binding is able to induce progressively growing colonies of NRK cells in soft agar, even without additional EGF. 相似文献
3.
Eric W. Johnson Susan F. Meunier Christopher J. Roy Nancy L. Parenteau 《In vitro cellular & developmental biology. Animal》1992,28(6):429-435
Summary We have developed a defined method for human epidermal keratinocyte culture. The minimally supplemented basal medium supported
establishment of primary cultures from neonatal foreskin in a defined environment. It also supported serial cultivation and
rapid expansion of cell number. Casein replaced serum for defined cryopreservation. Cells were serially cultivated in medium
containing 0.08 mM calcium. The rate of cell division however remained high after addition of 1.8 mM calcium. The particulate transglutaminase activity of the cultures was low at confluence, even in the presence of 1.88 mM calcium, indicating an enrichment of the basal cell population. Culture with small amounts (0.3%) of chelated serum increased
particulate transglutaminase activity approximately 2.2-fold in low calcium cultures and approximately 3.5-fold in high calcium
cultures. A gradual reduction in growth rate of serum-treated cultures upon serial cultivation also indicated a depletion
of cells with basal cell character. Bovine hypothalamic extract and cholera toxin were able to avert, in part, the differentiation-promoting
effects of serum. Keratinocytes serially cultivated in the defined medium maintained the ability to develop normally into
a morphologically differentiated epidermis. 相似文献
4.
Rapid and reversible changes in myosin heavy chain expression in response to increased neuromuscular activity of rat fast-twitch muscle 总被引:1,自引:0,他引:1
Chronic 10 Hz stimulation of rat fast-twitch muscle induced rapid and reversible changes in the tissue levels of fast myosin heavy chain (HC) mRNA isoforms. These changes consisted of a rapid decrease in HCIIb mRNA and a progressive increase in HCIIa mRNA. After 15 days, the HCIIb mRNA normally amounting to approximately 80%, had decreased to less than 5% of the sum of the two HC mRNA isoforms. HCIIb mRNA was again detectable one day after cessation of stimulation and progressively increased at the expense of HCIIa mRNA with ongoing recovery. These results point to a down-regulation of the HCIIb gene by the applied stimulus pattern which, conversely, enhances the expression of the HCIIa gene. 相似文献
5.
Nucleotide sequence of noncoding regions in Rous-associated virus-2: comparisons delineate conserved regions important in replication and oncogenesis. 总被引:40,自引:29,他引:11
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Fujinami sarcoma virus (FSV) encodes a 140,000-dalton transforming protein, P140, which contains gag- and fps-specific sequences. The cellular localization of this protein was examined by fractionation of [35S]methionine-labeled, FSV-infected chicken embryo fibroblasts. In homogenates of cells infected by wild-type, temperature-resistant FSV prepared in either hypotonic or isotonic buffer, 60 to 80% of the P140 was particulate. Isopycnic separation on discontinuous sucrose gradients indicated that the majority of the particulate P140 was present in a light membrane fraction enriched for plasma membranes. Much of the particulate P140 could be solubilized by the addition of 0.6 M salt to a postnuclear supernatant, suggesting that P140 is not an integral membrane protein. Particulate P140 may be associated with membranes either directly as a peripheral membrane protein or indirectly via cytoskeletal elements. In cells infected by mutants of FSV temperature sensitive for cellular transformation, most of the P140 is particulate at the permissive temperature, whereas most is soluble at the nonpermissive temperature; this change in distribution is not a secondary consequence of the change in cellular phenotype, since it also occurs in nonconditionally transformed cells doubly infected with temperature-sensitive FSV and wild-type Rous sarcoma virus. The movement of P140 from the particulate to the soluble fraction occurs rapidly when cells infected by temperature-sensitive FSV are shifted from the permissive to the nonpermissive temperature. Furthermore, P140 moves from the soluble to the particulate fraction, although somewhat more slowly, when cells are shifted from the nonpermissive to the permissive temperature. These observations suggest that the association of P140 with plasma membranes or the cytoskeleton may play a role in transformation by FSV. 相似文献
6.
Although histone synthesis and DNA synthesis are normally very well coordinated in HeLa cells, their histone synthesis proved relatively resistant to inhibition by ionizing radiation. During the first 24 h after 1 000 R the rate of cellular DNA synthesis progressively fell to small fractions of control values while histone synthesis continued with much less relative reduction. Acrylamide gel electropherograms of the acid soluble nuclear histones synthesized by irradiated HeLa cells were qualitatively normal. 相似文献
7.
The transforming proteins of PRCII virus and Rous sarcoma virus form a complex with the same two cellular phosphoproteins 总被引:5,自引:2,他引:3
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P105 and P110, the presumptive transforming proteins of PRCII avian sarcoma virus, have been found to be present in transformed chicken cells in two forms: as monomers and as part of a complex which contains both a 50,000-dalton and a 90,000-dalton cellular phosphoprotein. The 90,000-dalton cellular protein was found to be identical to one of the proteins in chicken cells whose synthesis is induced by stress. The 50,000-dalton protein was found to contain phosphotyrosine when isolated from the complex and therefore may be a substrate for the tyrosine protein kinase activity which is associated with P105 and P110. These same two cellular phosphoproteins have previously been shown to be present in a complex with pp60src, the tyrosine protein kinase which is the transforming protein of Rous sarcoma virus. However, not all avian sarcoma virus transforming proteins with associated tyrosine protein kinase activities form a complex efficiently with these cellular proteins. Little if any of P90, the putative transforming protein of Yamaguchi 73 virus, was found in a complex with the 50,000-dalton and 90,000-dalton cellular phosphoproteins. 相似文献
8.
Unstable expression and amplification of a transfected oncogene in confluent and subconfluent cells. 总被引:7,自引:6,他引:1
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N Glanville 《Molecular and cellular biology》1985,5(6):1456-1464
NIH 3T3 cells were transfected with a plasmid containing the transforming gene, v-src, from Rous sarcoma virus. One of the transformed cell lines isolated reverted to a flat, nontransformed morphology after cloning through soft agar. This cell line did not express the src gene and could no longer grow in soft agar. When these cells were held at confluence, spontaneous foci appeared which eventually covered the dish. The appearance of foci correlated with an increase in v-src gene expression, ability to grow in soft agar, and tumorigenicity in mice. When these transformed cells were trypsinized and held at subconfluence, both v-src expression and the transformed phenotype were progressively lost. Whereas rearrangement of the transfected gene was not detected, the gene copy number in the transformed cells was markedly increased (greater than 50-fold). Confluence-dependent gene amplification and deamplification have been retained after several cycles of growth alternately at high and low density, in cells recloned through soft agar, and after cells had been maintained continuously at high or low density. The results suggest that, in this cell line, reversible gene amplification plays a central role in expression of the transfected gene. 相似文献
9.
H. Polioudaki M.-C. Kastrinaki† H. A. Papadaki† P. A. Theodoropoulos 《Cell proliferation》2009,42(4):434-447
Objectives: This study aimed to investigate molecular and cellular changes induced in human bone marrow mesenchymal stem cells (hMSCs) after treatment with microtubule-interacting agents and to estimate damage to the bone marrow microenvironment caused by chemotherapy.
Materials and methods: Using an in vitro hMSC culture system and biochemical and morphological approaches, we studied the effect of nocodazole and taxol® on microtubule and nuclear envelope organization, tubulin and p53 synthesis, cell cycle progression and proliferation and death of hMSCs isolated from healthy donors.
Results and conclusions: Both nocodazole and taxol reduced hMSC proliferation and induced changes in the microtubular network and nuclear envelope morphology and organization. However, they exhibited only a moderate effect on cell death and partial arrest of hMSCs at G2 but not at M phase of the cell cycle. Both agents induced expression of p53, exclusively localized in abnormally shaped nuclei, while taxol, but not nocodazole, increased synthesis of β-tubulin isoforms. Cell growth rates and microtubule and nuclear envelope organization gradually normalized after transfer, in drug-free medium. Our data indicate that microtubule-interacting drugs reversibly inhibit proliferation of hMSCs; additionally, their cytotoxic action and effect on microtubule and nuclear envelope organization are moderate and reversible. We conclude that alterations in human bone marrow cells of patients under taxol chemotherapy are transient and reversible. 相似文献
Materials and methods: Using an in vitro hMSC culture system and biochemical and morphological approaches, we studied the effect of nocodazole and taxol® on microtubule and nuclear envelope organization, tubulin and p53 synthesis, cell cycle progression and proliferation and death of hMSCs isolated from healthy donors.
Results and conclusions: Both nocodazole and taxol reduced hMSC proliferation and induced changes in the microtubular network and nuclear envelope morphology and organization. However, they exhibited only a moderate effect on cell death and partial arrest of hMSCs at G
10.
Bioavailability of different phosphorus forms in freshwater systems 总被引:26,自引:9,他引:17
The recent literature on the bioavailability of different forms of P in freshwater systems is reviewed. Bioavailable P is defined as the sum of immediately available P and the P that can be transformed into an available form by naturally occurring processes. Methods used to estimate the bioavailable P pool, which vary between studies largely depending on the time perspective applied, are critically evaluated. Most studies on particulate P aim to determine the potentially available P pool. Potential bioavailability of particulate P is normally analysed in bioassays with algal yield determinations and the available P fraction is characterized from interpretations of results of sequential chemical extractions. NaOH-extractable P is in most studies the most algal-available P fraction. For soil samples and tributary water particulate matter, NaOH-P has often been found to be equal to algal extractable P. In other studies depletions of NaOH-P have accounted for the algal P uptake, but only a minor proportion of the fraction has been utilized. Organic P in lake water particulate matter and bed sediments of eutrophic lakes can also be algal-available to a significant extent.Studies on the bioavailability of dissolved P have often been concerned with immediate availability, or the minimum amount of available P. Such studies need other types of experimental design and normally assays with radiotracers are used. Immediately available P is frequently found to be less than P chemically assessed as dissolved reactive P (DRP) at low (< 10 µg DRP·l-1) concentrations. However, immediate availability may also approach or exceed DRP concentrations, especially at higher concentrations. Potential bioavailability, assayed as for particulate P, may generally render higher bioavailability than P assayed as immediately available. Large fractions of dissolved P remain unutilized and are primarily found in the high molecular weight fraction of dissolved P. 相似文献
11.
J A Levy 《Biomedicine / [publiée pour l'A.A.I.C.I.G.]》1981,34(1):4-8
Type C RNA viruses have been considered oncogenic because they are found associated with animal tumors and can induce cancers in several animal species. Those viruses that rapidly cause cancer appear to contain an oncogenic gene which resembles genetic sequences present in normal cells. This gene codes for a transforming protein which may be a normal cellular enzyme or a slightly altered cellular product. Its mechanism for transforming a cell is not yet known. Other oncogenic viruses, such as the chronic leukemia viruses, may not produce an oncogenic protein but may affect, by other means, specific target cells so they become malignant. Recent evidence now suggests that the majority of endogenous type C viruses are not transforming agents but inherited in the host to function in other biologic processes. These viruses do not contain transduced cellular genes which are responsible for cancer. Their role probably depends on their expression of other gene products which aid in normal development. These observations suggest that the ultimate control of human cancer may result from the identification of the oncogenic cellular-like genes transduced by some type C viruses even if a true human oncogenic virus is not isolated. 相似文献
12.
Molecular events leading to enhanced glucose transport in Rous sarcoma virus-transformed cells 总被引:2,自引:0,他引:2
Transformation by Rous sarcoma virus results in a dramatic increase in the rate at which the transformed cells transport glucose across the cell membrane. The increased transport rate is a consequence of an increased number of transporters in the transformed cells. Utilizing antibody raised against the purified human erythrocyte glucose transporter, we have identified the glucose transporter as a membrane glycoprotein with a monomer Mr of approximately 41,000. The increased rate of glucose transport is dependent on the activity of pp60src, the transforming protein of Rous sarcoma virus. This protein has been shown to be a protein kinase that phosphorylates on tyrosine residues. We have examined the tyrosine phosphorylation of a major cellular protein of Mr 36,000 in cells infected with a panel of partially transforming mutants of Rous sarcoma virus. One of these mutants (CU2) increases the rate of glucose transport only slightly and does not render the infected cells fully anchorage independent or tumorigenic (although other transformation parameters are fully induced). Cells infected with this mutant display a 36,000-dalton protein that is phosphorylated to a considerably lesser extent than cells infected with wild-type virus. Analyses of this sort may help to identify the cellular targets of pp60src whose phosphorylation is necessary for the increased glucose transport rate. 相似文献
13.
14.
A strain-specific modifier on mouse chromosome 4 controls the methylation of independent transgene loci 总被引:22,自引:0,他引:22
A transgene, pHRD, is highly methylated in 12 independent mouse lines when in a C57BL/6 strain background, but becomes progressively less methylated when bred into a DBA/2 background. Transgenes inherited from the mother are generally more methylated; however, this parental effect disappears following continued breeding into the nonmethylating strain. Mapping experiments using BXD recombinant inbred mice as well as other inbred strains indicate that a single strain-specific modifier (Ssm-1) linked to, but distinct from, Fv-1 is responsible for the strain effect. In addition to the methylated and unmethylated transgenic phenotypes, certain mice exhibit a partial methylation pattern that is a consequence of an unusual cellular mosaicism. The pHRD transgene, containing target sequences for the V(D)J recombinase, undergoes site-specific recombination only in lymphoid tissues. This V-J joining is restricted primarily to unmethylated transgene copies. 相似文献
15.
We have previously shown that following psoralen photoactivation (PUVA treatment) human dermal fibroblasts undergo long-term growth arrest as well as morphological and functional changes reminiscent of cellular senescence [ 1 ]. In the absence of molecular data on what constitutes normal senescence, it has been difficult to decide whether these PUVA-induced changes reflect cellular senescence or rather a mimic thereof. We herein report that PUVA-induced growth arrest, the senescent phenotype with long-term induction of senescence-associated beta-galactosidase, as well as increased expression of matrix metalloprotease-1 are fully reversible at days 100 to 130 post PUVA treatment in four independently tested fibroblast strains. The late returning growth capacity in PUVA-treated fibroblasts is not due to immortalization, as shown by continued lack of telomerase activity, accelerated telomere shortening, and a decrease in overall growth rates in fibroblasts in their regrowing phase post PUVA treatment. Lack of anchorage-independent growth additionally suggests that the cells are also not tumorigenically transformed. Collectively, our data suggest that PUVA-induced changes do not fully reflect replicative senescence but rather represent a long-term transient phenocopy of senescence. The model reported here is particularly suited to elucidating mechanisms underlying long-term transient growth arrest, the related functional changes, and the release of cells thereof. 相似文献
16.
1. The inherited differential agglutinability of cattle erythrocytes is shown to be similarly expressed on ghosts and intact cells.
2. Removal of virtually all sialic acid by prolonged neuraminidase treatment does not alter the agglutinability status of ghosts prepared from either high or low agglutinable cells. Hence the differing sialic acid content of the two cell types is not responsible for the differential agglutinability.
3. The significance of these findings with respect to other well defined agglutination systems and current theories of membrane structure is discussed. 相似文献
2. Removal of virtually all sialic acid by prolonged neuraminidase treatment does not alter the agglutinability status of ghosts prepared from either high or low agglutinable cells. Hence the differing sialic acid content of the two cell types is not responsible for the differential agglutinability.
3. The significance of these findings with respect to other well defined agglutination systems and current theories of membrane structure is discussed. 相似文献
17.
Association of pp60src and src protein kinase activity with the plasma membrane of nonpermissive and permissive avian sarcoma virus-infected cells. 总被引:19,自引:6,他引:13
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The intracellular localization of pp60src and src protein kinase activity in avian sarcoma virus (ASV)-infected chicken embryo fibroblasts and transformed and morphologically reverted field vole cells was examined by subcellular fractionation procedures. Fractionation by differential centrifugation of Dounce-homogenized cellular extracts prepared from vole cells showed that 83 to 91% of pp60src sedimented with particulate subcellular components from both transformed and revertant vole cells. A slightly lesser amount (60 to 70%) of pp60src was found associated with the particulate fraction from ASV-infected chicken embryo fibroblasts. The distribution of src protein kinase activity in the cytosol and particulate cell fractions was identical to that of pp60src, indicating no detectable differences in the activity of cytosol- and particulate-associated pp60src. When subcellular components of the cell were fractionated by discontinuous sucrose gradient centrifugation, similar amounts of both pp60src and src protein kinase activity cosedimented with the plasma membrane fractions from both transformed and revertant vole cells, as well as from ASV-infected chicken embryo fibroblasts. src protein kinase activity associated with plasma membrane fractions prepared from vole cells and ASV-infected chicken embryo fibroblasts was resistant to extraction with high salt concentrations, but partial elution was achieved with nonionic detergent. Thus, in both transformed and morphologically reverted vole cells, pp60src is intimately associated with the plasma membrane. Since transforming virus can be rescued from revertant vole cells by fusion to chicken embryo fibroblasts, revertant vole cell pp60src is capable of inducing morphological transformation. Thus, although the data presented herein suggest that transformation requires the association of pp60src with the plasma membrane, the binding of pp60src to the plasma membrane per se is insufficient to induce morphological transformation and requires the additional interaction with a specific target membrane protein which appears to be defective in revertant vole cells. 相似文献
18.
The STP-C488 oncogene of herpesvirus saimiri has transforming activity independent of the rest of the viral genome. We now demonstrate that STP-C488 associates with cellular ras in transformed cells. Mutations that disrupted this association with ras disrupted the transforming ability of the STP-C488 oncogene. Binding assays showed that STP-C488 was capable of competing with raf-1 for binding to ras. Expression of STP-C488 activated the ras signaling pathway as evidenced by a two- to fourfold increase in the ratio of ras-GTP to ras-GDP and by the constitutive activation of mitogen-activated protein kinase. Consistent with an activation of signaling through ras, STP-C488 expression induced ras-dependent neurite outgrowth in PC12 cells. STP-C488 is the first virus-encoded protein shown to achieve oncogenic transformation via association with cellular ras. 相似文献
19.
Control of myogenic differentiation by cellular oncogenes 总被引:5,自引:0,他引:5
20.
M. E. N. MAJERUS 《Ecological Entomology》1981,6(4):417-422
Abstract. 1. A New Forest population of P.hippocastanaria with both typical and nigrescens forms was studied over 6 years. The nigrescens form constituted between 10% and 17% of the population.
2. Breeding experiments showed that the nigrescens form is inherited as a unifactorial recessive; however, rearing at low temperatures induces a phenotype similar to nigrescens to result from insects with a genotype which normally produces typical imagines.
3. Old collections of P.hippocastanaria suggest that the frequency of the nigrescens form in the New Forest area has increased over the last 100 years.
4. Samples collected from other areas, within and outside the New Forest, indicated that the distribution of nigrescens is limited to heaths within the New Forest and on the extreme eastern border of Dorset.
5. A number of speculative reasons for the increase in the frequency of the nigrescens form in this area are suggested. 相似文献
2. Breeding experiments showed that the nigrescens form is inherited as a unifactorial recessive; however, rearing at low temperatures induces a phenotype similar to nigrescens to result from insects with a genotype which normally produces typical imagines.
3. Old collections of P.hippocastanaria suggest that the frequency of the nigrescens form in the New Forest area has increased over the last 100 years.
4. Samples collected from other areas, within and outside the New Forest, indicated that the distribution of nigrescens is limited to heaths within the New Forest and on the extreme eastern border of Dorset.
5. A number of speculative reasons for the increase in the frequency of the nigrescens form in this area are suggested. 相似文献