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1.
Cell growth and tRNA-lys4 synthesis in mouse 3T3 cells 总被引:2,自引:0,他引:2
B J Ortwerth J Wolters J Nahlik C Conlon-Hollingshead 《Experimental cell research》1982,138(2):241-250
The RPC-5 chromatographic profiles of lys-tRNA were analyzed during the growth of 3T3 cells in culture. An inverse relationship was seen between tRNA2lys and tRNA4lys which was markedly influenced by medium changes. This interchange of tRNA2lys and tRNA4lys could be controlled by altering the levels of serum in the medium, or more precisely by altering the serum to cell ratio. A different change in lys-tRNA distribution was seen when the cells reached confluency. The amounts of tRNA2lys, tRNA3lys and tRNA4lys all decreased with a corresponding increase in either tRNA5lys or tRNA6lys. An identical change in lys-tRNA could be produced by shifting sparse cells into a medium containing 10% calf plasma instead of 10% serum. Both tRNAlys profiles and cell growth were returned to normal when the cells were returned to medium with 10% fetal calf serum (FCS) or 10% calf plasma and fibroblast growth factor (FGF). A third alteration in tRNAlys profiles was seen by the addition of cAMP to the cultures. A decrease in tRNA5lys and a corresponding increase in tRNA6lys was seen upon the addition of 10?3 M db-cAMP and was accentuated by the simultaneous addition of 10?3 M methyl isobutylxanthine.These data are consistent with an ordered sequence of tRNAlys modification involving tRNA2lys, tRNA3lys, tRNA4lys, tRNA5Blys and tRNA6lys. Several of the factors which control proliferation appear to control the activity of different tRNA-modifying enzymes in this tRNAlys pathway thereby controlling the levels of tRNA4lys, a tRNA previously shown to correlate directly with the proliferative rate of cells. 相似文献
2.
B. J. Ortwerth J. Wolters J. Nahlik Christine Conlon-Hollingshead 《Experimental cell research》1982,138(2)
The RPC-5 chromatographic profiles of lys-tRNA were analyzed during the growth of 3T3 cells in culture. An inverse relationship was seen between tRNA2lys and tRNA4lys which was markedly influenced by medium changes. This interchange of tRNA2lys and tRNA4lys could be controlled by altering the levels of serum in the medium, or more precisely by altering the serum to cell ratio. A different change in lys-tRNA distribution was seen when the cells reached confluency. The amounts of tRNA2lys, tRNA3lys and tRNA4lys all decreased with a corresponding increase in either tRNA5lys or tRNA6lys. An identical change in lys-tRNA could be produced by shifting sparse cells into a medium containing 10% calf plasma instead of 10% serum. Both tRNAlys profiles and cell growth were returned to normal when the cells were returned to medium with 10% fetal calf serum (FCS) or 10% calf plasma and fibroblast growth factor (FGF). A third alteration in tRNAlys profiles was seen by the addition of cAMP to the cultures. A decrease in tRNA5lys and a corresponding increase in tRNA6lys was seen upon the addition of 10−3 M db-cAMP and was accentuated by the simultaneous addition of 10−3 M methyl isobutylxanthine.These data are consistent with an ordered sequence of tRNAlys modification involving tRNA2lys, tRNA3lys, tRNA4lys, tRNA5Blys and tRNA6lys. Several of the factors which control proliferation appear to control the activity of different tRNA-modifying enzymes in this tRNAlys pathway thereby controlling the levels of tRNA4lys, a tRNA previously shown to correlate directly with the proliferative rate of cells. 相似文献
3.
Santa Spaggiare Michael J. Wallach Joseph T. Tupper 《Journal of cellular physiology》1976,89(3):403-416
The components of unidirectional K influx and efflux have been investigated in the 3T3 cell and the SV40 transformed 3T3 cell in exponential and stationary growth phase. Over the cell densities used for transport experiments the 3T3 cell goes from exponential growth to density dependent inhibition of growth (4 × 104 to 4 × 105 cell cm?2) whereas the SV40 3T3 maintains exponential or near exponential growth (4 × 104 to 1 × 106 cell cm?2). In agreement with previous observations, volume per cell and mg protein per cell decrease with increasing cell density. Thus, transport measurements have been expressed on a per volume basis. Total unidirectional K influx and efflux in the 3T3 cell is approximately double that of the SV40 3T3 cell at all cell densities investigated. Both cell types have similar volumes initially and show similar decreases with increasing cell density. Thus, in this clone of the 3T3 cell SV40 transformation specifically decreases unidirectional K flux. The magnitude of the total K flux does not change substantially for either cell line during transition from sparse to dense cultures. However, the components of the K transport undergo distinct changes. Both cell lines possess a ouabain sensitive component of K influx, presumably representing the active inward K pump. Both also possess components of K influx and efflux sensitive to furosemide. The data suggest this component represents a one-for-one K exchange mechanism. The fraction of K influx mediated by the ouabain sensitive component is reduced to one half its value when exponential versus density inhibited 3T3 cells are compared (63% versus 31% of total influx). No comparable drop occurs in the SV40 3T3 cell at equivalent cell densities (64% versus 56% of total influx). Thus, the pump mediated component of K influx would appear to be correlated with growth. In contrast, the furosemide sensitive component represents approximately 20% of the total unidirectional K influx and efflux in both cell lines in sparse culture. At high cell densities, where growth inhibition occurs in the 3T3 cell but not the SV40 3T3, the furosemide sensitive component doubles in both cell lines. Thus, the apparent K-K exchange mechanism is density dependent rather than growth dependent. 相似文献
4.
Biochemical characterization of the mitochondrial tRNASer(UCN) T7511C mutation associated with nonsyndromic deafness 总被引:6,自引:0,他引:6
Li X Fischel-Ghodsian N Schwartz F Yan Q Friedman RA Guan MX 《Nucleic acids research》2004,32(3):867-877
We report here the biochemical characterization of the deafness-associated mitochondrial tRNASer(UCN) T7511C mutation, in conjunction with homoplasmic ND1 T3308C and tRNAAla T5655C mutations using cybrids constructed by transferring mitochondria from lymphoblastoid cell lines derived from an African family into human mtDNA-less (ρ°) cells. Three cybrids derived from an affected matrilineal relative carrying the homoplasmic T7511C mutation, exhibited ~75% decrease in the tRNASer(UCN) level, compared with three control cybrids. This amount of reduction in the tRNASer(UCN) level is below a proposed threshold to support a normal rate of mitochondrial protein synthesis in lymphoblastoid cell lines. This defect is likely a primary contributor to ~52% reduction in the rate of mitochondrial protein synthesis and marked defects in respiration and growth properties in galactose-containing medium. Interestingly, the T5655C mutation produces ~50% reduction in the tRNAAla level in mutant cells. Strikingly, the T3308C mutation causes a significant decrease both in the amount of ND1 mRNA and co-transcribed tRNALeu(UUR) in mutant cells. Thus, mitochondrial dysfunctions caused by the T5655C and T3308C mutations may modulate the phenotypic manifestation of the T7511C mutation. These observations imply that a combination of the T7511C mutation with two mtDNA mutations accounts for the high penetrance of deafness in this family. 相似文献
5.
The Deafness-Associated Mitochondrial DNA Mutation at Position 7445, Which Affects tRNASer(UCN) Precursor Processing, Has Long-Range Effects on NADH Dehydrogenase Subunit ND6 Gene Expression 总被引:9,自引:1,他引:8
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Min-Xin Guan Jos Antonio Enriquez Nathan Fischel-Ghodsian Ram S. Puranam Catherine P. Lin Marion A. Maw Giuseppe Attardi 《Molecular and cellular biology》1998,18(10):5868-5879
The pathogenetic mechanism of the deafness-associated mitochondrial DNA (mtDNA) T7445C mutation has been investigated in several lymphoblastoid cell lines from members of a New Zealand pedigree exhibiting the mutation in homoplasmic form and from control individuals. We show here that the mutation flanks the 3′ end of the tRNASer(UCN) gene sequence and affects the rate but not the sites of processing of the tRNA precursor. This causes an average reduction of ~70% in the tRNASer(UCN) level and a decrease of ~45% in protein synthesis rate in the cell lines analyzed. The data show a sharp threshold in the capacity of tRNASer(UCN) to support the wild-type protein synthesis rate, which corresponds to ~40% of the control level of this tRNA. Strikingly, a 7445 mutation-associated marked reduction has been observed in the level of the mRNA for the NADH dehydrogenase (complex I) ND6 subunit gene, which is located ~7 kbp upstream and is cotranscribed with the tRNASer(UCN) gene, with strong evidence pointing to a mechanistic link with the tRNA precursor processing defect. Such reduction significantly affects the rate of synthesis of the ND6 subunit and plays a determinant role in the deafness-associated respiratory phenotype of the mutant cell lines. In particular, it accounts for their specific, very significant decrease in glutamate- or malate-dependent O2 consumption. Furthermore, several homoplasmic mtDNA mutations affecting subunits of NADH dehydrogenase may play a synergistic role in the establishment of the respiratory phenotype of the mutant cells. 相似文献
6.
The effects of cytochalasin B, vinblastine and temperature on particle movement in the plasma membrane of several 3T3 mouse fibroblast lines were investigated. Preincubation of normal 3T3 cells for 24 h in 5–10 μg/ml cytochalasin B had no effect on the mean square relative displacement of marker particles in the membrane (motion constant), but preincubation for 4 h in 40 μg/ml vinblastine reduced the value of this constant by 70%. A 10 °C decrease in temperature decreased the motion constant in normal cells (Q10 = 4) more than in virus-transformed 3T3 cells (Q10 = 1.8). Interpreting the motion constant of the particles as an expression of the viscosity of the membrane material, values of 3 poise for normal 3T3 cells and 6 poise for the transformed cells are obtained for 37 °C and pH = 7.4.A method is suggested to quantitate aggregation of particles on the surface of cells. When this method is applied to gold particles on 3T3 cells, disaggregation of particles is seen to behave as an unordered process, whereas aggregation appears to express cellular control. This consideration and the effect of vinblastine indicate that the interpretation of particle movement as Brownian movement in a viscous membrane material does not cover all phenomena observed.The membrane movement of the flat revertant SVF1 101 [1] was investigated. This cell line occupies an intermediary position between normal 3T3 mouse fibroblasts and the polyoma and SV40 transformed 3T3 cell lines as judged by growth properties. Its membrane movement was found to occupy an intermediary position between the membrane movements of these cell lines, too. 相似文献
7.
Melanie A. Preston Sonia D’Silva Yoshiko Kon Eric M. Phizicky 《RNA (New York, N.Y.)》2013,19(2):243-256
tRNAs are highly modified, each with a unique set of modifications. Several reports suggest that tRNAs are hypomodified or, in some cases, hypermodified under different growth conditions and in certain cancers. We previously demonstrated that yeast strains depleted of tRNAHis guanylyltransferase accumulate uncharged tRNAHis lacking the G−1 residue and subsequently accumulate additional 5-methylcytidine (m5C) at residues C48 and C50 of tRNAHis, due to the activity of the m5C-methyltransferase Trm4. We show here that the increase in tRNAHis m5C levels does not require loss of Thg1, loss of G−1 of tRNAHis, or cell death but is associated with growth arrest following different stress conditions. We find substantially increased tRNAHis m5C levels after temperature-sensitive strains are grown at nonpermissive temperature, and after wild-type strains are grown to stationary phase, starved for required amino acids, or treated with rapamycin. We observe more modest accumulations of m5C in tRNAHis after starvation for glucose and after starvation for uracil. In virtually all cases examined, the additional m5C on tRNAHis occurs while cells are fully viable, and the increase is neither due to the GCN4 pathway, nor to increased Trm4 levels. Moreover, the increased m5C appears specific to tRNAHis, as tRNAVal(AAC) and tRNAGly(GCC) have much reduced additional m5C during these growth arrest conditions, although they also have C48 and C50 and are capable of having increased m5C levels. Thus, tRNAHis m5C levels are unusually responsive to yeast growth conditions, although the significance of this additional m5C remains unclear. 相似文献
8.
The role of mitogen-activated calcium influx from the extracellular medium in the control of cell proliferation was studied in Balb-c 3T3 fibroblasts. Stimulation of serum-deprived, quiescent cells with 10% foetal calf serum (FCS) induced a long-lasting (up to 70 min elevation of intracellular free calcium concentration ([Ca2+]i). Both the sustained [Ca 2+]i increase and the related inward current, described in a previous paper [Lovisolo D. Munaron L. Baccino FM. Bonelli G. (1992) Potassium and calcium currents activated by foetal calf serum in Balb-c 3T3 fibroblasts. Biochim. Biophys. Acta, 1104, 73–82], could be abolished either by chelation of extracellular calcium with EGTA or by SKF 96365, an imidazole derivative that can block receptor-activated calcium channels. The effect of the abolition of these ionic signals on FCS-induced proliferation was investigated by adding either EGTA or SK&F 96365 to the culture medium during the first hours of stimulation of quiescent cells with 10% FCS. As measured after 24 h, a 22% inhibition of growth was observed when SK&F 96365 was added for the first hour, and stronger inhibitions, up to 56%, were obtained by adding the blocker for the first 2 or 4 h. Similar effects were observed with addition of 3 mM EGTA, though the inhibition was less marked for the 4 h treatment. By contrast, incubation with either substance in the next 4 h of serum stimulation did not influence cell growth, except for a slight inhibition observed when SKF 96365 was applied from the 4th to the 8th hour. The reduction in growth resulting from the abolition of the early calcium influx was paralleled by an accumulation of cells in the G2/M phase. Both growth inhibition and G2/M accumulation were reversible, since after further 24 h in 10% FCS cells had fully recovered the exponential growth. These data indicate that the early calcium influx seen in response to mitogen stimulation develops on a timescale long enough to play a significant role in cell cycle progression, and that its block in the early G1 phase can lead to a reduction of proliferation by arresting cells in later stages of the cycle. 相似文献
9.
The cloned C3H/10T1/2 mouse embryo cells contained a complex pattern of gangliosides. Two cloned chemical transformants obtained from the C3H/10T1/2 cell line by treatment with 7,12-dimethylbenz(a) anthracene (DMBA-TCL1) and 3-methylcholanthrene (MCA-TCL15) also had complex ganglioside patterns; but the transformants had increased levels of the simplest ganglioside, N-acetylneuraminylgalactosylglucosylceramide (GM3), and reduced levels of more complex gangliosides. Incorporation of [14C]glucosamine into gangliosides, as cell-to-cell contact increased in C3H/10T1/2 cells, showed that GM3 synthesis was decreased and that the synthesis of the more complex ganglioside N-acetylneuraminylgalactosyl-N-acetylgalactosaminyl-(N-acetylneuraminyl)-galactosylglucosylceramide (GD1a) was increased. In the two transformants the percentage each individual ganglioside was of total labeled gangliosides was only slightly altered with changing cell density. Turnover of [14C]glucosamine-labeled gangliosides, as cell density increased, was approximately equal in C3H/10T1/2 cells and MCA-TCL15 cells, but more rapid in the DMBA-TCL1 cells. Most individual gangliosides turned over at about the same rate in the respective cell lines. However, GD1a increased slightly as a percentage of total labeled gangliosides with increasing cell density in both C3H/10T1/2 cells and transformed cells. The labeling data indicated that the majority of GD1a synthesis was de novo and only a small part occurred by transfer of sialyl or glycosyl residues to simpler gangliosides or catabolism of more complex gangliosides already present in the outer membrane. Exogenous complex gangliosides added to the medium were more effective inhibitors of DMBA-TCL1 cell growth than of C3H/10T1/2 cell growth. Furthermore, gangliosides added to exponentially growing C3H/10T1/2 and DMBA-TCL1 cells caused both cell lines to incorporate a greater percentage of [14C]glucosamine into gangliosides more complex than GM3. 相似文献
10.
W H McClain 《Biochemical and biophysical research communications》1979,86(3):718-724
Synthesis of T4 tRNAGln depends on normal levels of ribonuclease III. Infection of cell strains carrying a mutation in the gene for this enzyme resulted in severe depression in tRNAGln production, as revealed by chemical and suppressor tRNA analyses. The remaining seven T4 tRNAs were synthesized in the mutant cells. The requirement of ribonuclease III for synthesis of tRNAGln points to an essential cleavage by the enzyme of a precursor RNA containing tRNAGln. 相似文献
11.
The uptake of ouabain-sensitive 86Rb+ uptake measured at 5 min and the uptake measured at 60 min was 4.5- and 2.7-fold greater respectively for SV40 transformed 3T3 cells compared to 3T3 cells during the late log phase of growth. This uptake, however, varied markedly with cell growth. Ouabain-sensitive 86Rb+ uptake was found to be a sensitive indicator of protein synthesis as measured by total protein content. Cessation of cell growth as measured by total protein content was associated with a decline in ouabain-sensitive 86Rb+ uptake in both cell types. This increased ouabain-sensitive cation transport was reflected in increased levels of (Na+ + K+)-ATPase activity for SV40 3T3 cells, which showed a 2.5-fold increase V but the same Krmm as 3T3 cells.These results are compared with the results of related work. Possible mechanisms for these effects are discussed and how changes in cation transport might be related to alterations in cell growth. 相似文献
12.
Diego Rojas-Benitez Patrick C. Thiaville Valérie de Crécy-Lagard Alvaro Glavic 《The Journal of biological chemistry》2015,290(30):18699-18707
N6-Threonylcarbamoyl-adenosine (t6A) is a universal modification occurring at position 37 in nearly all tRNAs that decode A-starting codons, including the eukaryotic initiator tRNA (tRNAiMet). Yeast lacking central components of the t6A synthesis machinery, such as Tcs3p (Kae1p) or Tcs5p (Bud32p), show slow-growth phenotypes. In the present work, we show that loss of the Drosophila tcs3 homolog also leads to a severe reduction in size and demonstrate, for the first time in a non-microbe, that Tcs3 is required for t6A synthesis. In Drosophila and in mammals, tRNAiMet is a limiting factor for cell and animal growth. We report that the t6A-modified form of tRNAiMet is the actual limiting factor. We show that changing the proportion of t6A-modified tRNAiMet, by expression of an un-modifiable tRNAiMet or changing the levels of Tcs3, regulate target of rapamycin (TOR) kinase activity and influences cell and animal growth in vivo. These findings reveal an unprecedented relationship between the translation machinery and TOR, where translation efficiency, limited by the availability of t6A-modified tRNA, determines growth potential in eukaryotic cells. 相似文献
13.
Strains of Escherichia coli have been produced which express very high levels of the tRNAleu1 isoacceptor. This was accomplished by transforming cells with plasmids containing the leuV operon which encodes three copies of the tRNALeu1 gene. Most transformants grew very slowly and exhibited a 15-fold increase in cellular concentrations of tRNALeu1 As a result, total cellular tRNA concentration was approximately doubled and 56% of the total was tRNALeu1. We examined a number of parameters which might be expected to be affected by imbalances in tRNA concentration: in vivo tRNA charging levels, misreading, ribosome step time, and tRNA modification. Surprisingly, no increase in intracellular ppGpp levels was detected even though only about 40% of total leucyl tRNA was found to be charged in vivo. Gross ribosomal misreading was not detected, and it was shown that ribosomal step times were reduced between two- and threefold. Analyses of leucyl tRNA isolated from these slow-growing strains showed that at least 90% of the detectable tRNALeu1 was hypomodified as judged by altered mobility on RPC-5 reverse-phase columns, and by specific modification assays using tRNA(m1G)-methyltransferase and pseudo-uridylate synthetase. Analysis of fast-growing revertants demonstrated that tRNA concentration per se may not explain growth inhibition because selected revertants which grew at wild-type growth rates displayed levels of tRNA comparable to that of control strains bearing the leuV operon. A synthetic tRNALeu1 operon under the control of the T7 promoter was prepared which, when induced, produced six- to sevenfold increases in tRNALeu1 levels. This level of tRNALeu1 titrated the modification system as judged by RPC-5 column chromatography. Overall, our results suggest that hypomodified tRNA may explain, in part, the observed effects on growth, and that the protein-synthesizing system can tolerate an enormous increase in the concentration of a single tRNA. 相似文献
14.
Joanne D. Elligsen J. E. Thompson H. E. Frey J. Kruuv 《Experimental cell research》1974,87(2):233-240
The (Na+---K+)-stimulated Mg2+-dependent ATPase activities of 3T3 and SV40 transformed 3T3 cells were compared as a function of cell population density. For normal cells the enzyme activity remained relatively constant during exponential growth, but decreased sharply coincident with contact inhibition of growth at confluence. This decrease in activity could be reversed by stimulating contact-inhibited cultures to undertake renewed short-term growth either by adding fetal calf serum or changing the medium completely. Transformed cells did not experience a decrease in (Na+---K+)-ATPase activity upon reaching confluence, but this is consistent with the fact that they were still growing exponentially at this stage. However, non-confluent cultures of both normal and transformed cells incurred a marked decrease in levels of the enzyme when growth was inhibited by serum depletion. The results have been interpreted as indicating that levels of (Na+---K+)-ATPase in both normal and transformed cells are correlated with growth. Hence the different patterns of ATPase activity displayed by malignant cells and their normal counterparts with increase in cell number appear to be a reflection of their dissimilar growth behaviours rather than of any innate difference between them. 相似文献
15.
Fumiyoshi Okano Hirofumi Tachibana Koichi Akiyama Sanetaka Shirahata Hiroki Murakami 《Cytotechnology》1993,11(3):205-211
Immortalized human T cell lines were established by cotransfecting c-Ha-ras and c-myc oncogenes to lymph node lymphocytes. The cell lines kept growing for 3 months after establishment without a decrease in growth rate. The cells did not require interleukin-2(IL-2) for their growth, but addition of IL-2 stimulated the growth of these cells. Flow cytometric analysis revealed that these cells were T cells expressing CD4 or CD8 antigens. A CD4 positive (CD4+) cell line produced IL-6, indicating that the cell line belongs to helper T cells. The CD8 positive (CD8+) cell line possessed cytotoxicity to tumor cells, indicating that the cell line were killer T cells. Both cell lines were able to proliferate in serum-free medium indefinitely. 相似文献
16.
Bau-Shan HsiehLi-Wen Huang Shu-Jem SuHsiao-Ling Cheng Yu-Chen HuThu-Ching Hung Kee-Lung Chang 《The Journal of nutritional biochemistry》2011,22(3):234-241
Arginine is a physiological substrate for nitric oxide synthase to generate nitric oxide (NO), which can influence tumor cell survival, while ascorbic acid is selectively toxic for cancer cells. This study explored the effect of an arginine/ascorbic acid combination on human cancer cell lines. The hepatoma cell line HA22T/VGH was the most sensitive of the tested cells to combination treatment. A combination of 5.74 mM of arginine and 0.57 mM of ascorbic acid induced HA22T/VGH cell death through apoptosis and an increase in levels of reactive oxygen species and NO, as well as its stable products NO2− and NO3−. The combination also reduced the activity of glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and transaldolase in the pentose phosphate pathway, a major mechanism for producing NADPH, resulting in a marked decrease in intracellular NADPH levels. A dramatic decrease in intracellular glutathione (GSH) levels, a decrease in the mitochondrial membrane potential, ATP depletion and release of cytochrome c were also seen. Caspase-9 and caspase-3 were activated, apoptotic protein Bax expression increased and the expression of the anti-apoptotic proteins Bcl-2 and Bcl-xL decreased. These results suggest that this combination induced HA22T/VGH cell death by interfering with redox state regulation by a reduction in pentose phosphate pathway activity and increasing oxidative and nitrosative stress. 相似文献
17.
Background
Bispecific T cell engager (BiTE®) are single-chain bispecific antibody constructs with dual specificity for CD3 on T cells and a surface antigen on target cells. They can elicit a polyclonal cytotoxic T cell response that is not restricted by T cell receptor (TCR) specificity, and surface expression of MHC class I/peptide antigen complexes. Using human EpCAM/CD3-bispecific BiTE® antibody construct AMG 110, we here assessed to what extent surface expression of PD-L1, cytoplasmic expression of indoleamine-2,3-deoxygenase type 1, Bcl-2 and serpin PI-9, and the presence of transforming growth factor beta (TGF-β), interleukin-10 (IL-10) and adenosine in culture medium can impact redirected lysis by AMG 110-engaged T cells.Methods
The seven factors, which are all involved in inhibiting T cell functions by cancer cells, were tested with human EpCAM-expressing Chinese hamster ovary (CHO) target cells at levels that in most cases exceeded those observed in a number of human cancer cell lines. Co-culture experiments were used to determine the impact of the evasion mechanisms on EC50 values and amplitude of redirected lysis by AMG 110, and on BiTE®-induced proliferation of previously resting human peripheral T cells.Findings
An inhibitory effect on redirected lysis by AMG 110-engaged T cells was seen upon overexpression of serpin PI-9, Bcl-2, TGF-βand PD-L1. An inhibitory effect on induction of T cell proliferation was only seen with CHO cells overexpressing IDO. In no case, a single evasion mechanism rendered target cells completely resistant to BiTE®-induced lysis, and even various combinations could not.Conclusions
Our data suggest that diverse mechanisms employed by cancer cells to fend off T cells cannot inactivate AMG 110-engaged T cells, and that inhibitory effects observed in vitro may be overcome by increased concentrations of the BiTE® antibody construct. 相似文献18.
R Dunn S Hayashi I C Gillam A D Delaney G M Tener T A Grigliatti T C Kaufman D T Suzuki 《Journal of molecular biology》1979,128(3):277-287
The genes for tRNA3bval were localized to 84D and 92B on the polytene chromosomes of Drosophila melanogaster with a possible minor site at 90B-C by hybridization in situ and autoradiography with 125I-labeled tRNA3bval. Flies carrying a duplication of the 84D region had increased amounts (30%) of tRNA3bval in proportion to the increased number of genes. While a proportional decrease in the amount of tRNAval3b in flies bearing a deletion of the same region was found, the total acceptance of valine remained at the level found in the wild type. 相似文献
19.
Expressions of polypeptide: N-acetylgalactosaminyltransferase in leukemia cell lines during 1,25-dihydroxyvitamin D3 induced differentiation 总被引:1,自引:0,他引:1
The effect of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] on two leukemia cell lines, K562 and SHI-1, and its relation to the expression of different subtypes of polypeptide: N-acetylgalactosaminyltransferase (pp-GalNAc-T) was studied. With morphological and cell flow-cytometric method, it was found
that 1,25(OH)2D3 induced the differentiation of both leukemia cell lines toward monocytic lineage, but not affected the cell growth and
apoptosis. The expressions of different subtypes of pp-GalNAc-T, the initial glycosyltransferase in O-glycan synthesis, were studied with RT-PCR before and after the treatment of different concentrations of 1,25(OH)2D3. Among fourteen subtypes of pp-GalNAc-T (T1 ∼ T14), K562 cells obviously expressed pp-GalNAc-T2, T4, T5, T7 (T2 was the
highest) and SHI-1 cells apparently expressed pp-GalNAcT1, T2, T3 and T4 (T4 was the highest) only. After K562 cells were
treated 1, 25(OH)2D3 for 72 h, pp-GalNAc-T2, T4, T5, T7 were increased in a dose dependent manner. In contrast, pp-GalNAc-T1 and T2, especially
T1, were up-regulated in SHI-1 cells by 1,25(OH)2D3, but T3 was unchanged and T4 was down-regulated. The different alterations of pp-GalNAc-Ts in these two cell lines were
probably related to the different structural changes of O-glycans during 1,25(OH)2D3 induced differentiation. 相似文献
20.
3T3 and SV-40 transformed 3T3 mouse fibroblasts were cultured in media with serum and antibiotics plus ammonia (NH3 z NH4+) added as NH4C1. Both cell lines cultured without added ammonia showed normal morphology and multiplication even though ammonia in the medium at the end of the culture period ranged from 35 to 48 μg/ml. Ammonia concentrations being significantly higher in media removed from cells at the end of the culture period than in media incubated identically without cells, verified that cells released substantial quantities of ammonia in addition to components of the medium which underwent spontaneous breakdown. Both cell lines showed changes in morphology and highly significant reductions in cell multiplication which increased progressively as the concentration of added ammonia on the initial day of culture was increased to 35μg/ml. Control 3T3 cultures released significantly greater quantities of ammonia per cell than control cultures of transformed cells but their multiplication was more adversely affected by added ammonia. There were downward shifts in pH of the culturing medium for both cell lines as culture age increased at all concentrations of added ammonia, However, significant reductions in cell multiplication resulted from additions of ammonia that did not produce significant changes in extracellular pH. The data show that studies upon the effects of pH of the medium on cultured cells require control of ammonia concentrations. 相似文献