GROWTH DEPENDENT 2-DEOXY-D-GLUCOSE UPTAKE IN TRANSFORMED AND NORMAL MOUSE CELLS

1974 ◽  
Vol 16 (4) ◽  
pp. 823-830 ◽  
Author(s):  
Patricia A. Martin-De Leon ◽  
Barid B. Mukherjee
Keyword(s):  
Fold Increase ◽  
Lung Fibroblasts ◽  
Transformed Cells ◽  
Embryonic Cells ◽  
Transformed Cell ◽  
Serum Stimulation ◽  
Dependent Inhibition ◽  
Sarcoma Virus ◽  
Cell Densities ◽  
Transformed Cell Lines

The rates of uptake of 2-deoxyglucose were studied in a) 15-day BALB/c and AKR embryonic cells, b) adult BALB/c lung fibroblasts and c) virally-transformed cell lines derived from a clone of BALB/c cells. At high cell densities in confluent cultures of non-transformed cells the rates of uptake were three to seven times lower than those in transformed cells at the equivalent densities. Both non-transformed and virally-transformed cell cultures showed density-dependent inhibition of 2-deoxyglucose uptake. However, inhibition was only slight in SV40 (SVT2) and Kirsten sarcoma virus-transformed (K-A31) cells in which uptake rates decreased 2.3– and 2.5– fold respectively during a 12-fold increase in cell densities. For an almost similar increase in cell density, in unestablished adult embryonic BALB/c fibroblasts, the rate of uptake decreased 8-fold. Incorporation of H3-thymidine (an indication of growth rate) occurred at a higher rate in transformed than in contact-inhibited cells. Two hours after serum stimulation of contact-inhibited AKR cells there was approximately 50% increase in the rate of 2-deoxyglucose uptake. The results confirm previous findings that the increased uptake of 2-deoxyglucose by virally-transformed cells may be due to their growth and division and not necessarily to the direct expression of the viral genome.

scholarly journals Regulation of proline 3-hydroxylation and prolyl 3-hydroxylase and 4-hydroxylase activities in transformed cells

1982 ◽  
Vol 206 (3) ◽  
pp. 499-503 ◽  
Author(s):  
K Majamaa ◽  
R Myllylä ◽  
K Alitalo ◽  
A Vaheri
Keyword(s):  
Enzyme Activity ◽  
Cell Lines ◽  
Enzyme Activities ◽  
Cell Transformation ◽  
Transformed Cells ◽  
Temperature Sensitive ◽  
Permissive Temperature ◽  
Hydroxylase Activity ◽  
Transformed Cell ◽  
Transformed Cell Lines

Prolyl 3-hydroxylase activity and the extent of collagen proline 3-hydroxylation were studied in six transformed and three control human cell lines. In the transformed cell lines, the enzyme activity was markedly high in two, similar to that in control cells in two and significantly low in two. The extent of proline 3-hydroxylation was markedly high in cell lines with high enzyme activity, but it was also significantly high in some transformed cell lines with enzyme activities similar to those in the controls. The results thus suggest that, in addition to the amount of enzyme activity present, the rate of collagen synthesis also affects the extent of proline 3-hydroxylation in the newly synthesized collagen. The effect of acute cell transformation on prolyl 3-hydroxylase and 4-hydroxylase activities was studied by infecting chick-embryo fibroblasts with Rous sarcoma virus mutant NY68, temperature-sensitive for transformation. At the permissive temperature prolyl 3-hydroxylase activity showed a more rapid increase and decrease than did prolyl 4-hydroxylase activity, the maximal activity for both enzymes being about 2.5 times that in the control chick fibroblasts. When the transformed cells were shifted to the non-permissive temperature the decays in the elevated enzyme activities were similar, suggesting identical half-lives.

scholarly journals Intracellular ionic changes in normal and transformed human fibroblasts after extracellular Ca2+ deprivation

1981 ◽  
Vol 194 (3) ◽  
pp. 707-711 ◽  
Author(s):  
B J Hazelton ◽  
J T Tupper
Keyword(s):  
Growth Response ◽  
Human Fibroblasts ◽  
Growth Arrest ◽  
Fold Increase ◽  
Transformed Cells ◽  
Transformed Cell ◽  
Permeability Changes ◽  
Na Efflux ◽  
Ionic Changes ◽  
G0 Phase

The lowering of extracellular Ca2+ concentration in the growth medium reversibly blocks normal, but not SV40-transformed WI38 diploid fibroblasts in the early G1/G0 phase of the cell cycle. This growth response is characterized by specific changes in ionic content and transport. Ca2+ deprivation (0.03 mM) has little effect on the K+ content of either normal or transformed cells. Na+ content, however, is increased nearly 2-fold in the normal cells. This increase is presumably due to a 3-fold increase in unidirectional Na+ influx in Ca2+-deprived cells. The increased intracellular Na+ also gives rise to a nearly 3-fold enhancement of the active (ouabain-sensitive) Na+ efflux. Ca2+ deprivation causes only slight increases in Na+ influx, ouabain-sensitive Na+ efflux and intracellular Na+ in the transformed cell. In contrast, the transformed cells lose nearly 60% of their intracellular Ca2+ on deprivation, whereas normal WI38 cells lose only 10%. The data suggest that the growth arrest exhibited by the normal cell but not the transformed cell may be related to different membrane-transport and permeability changes in response to Ca2+ deprivation.

Interaction Between Normal and Transformed Bovine Fibroblasts in Culture

1967 ◽  
Vol 2 (3) ◽  
pp. 309-322
Author(s):  
ELIZABETH MACINTYRE ◽  
J. PONTÉN
Keyword(s):  
Rous Sarcoma Virus ◽  
Lung Fibroblasts ◽  
Transformed Cells ◽  
Embryonic Lung ◽  
Normal Cells ◽  
Stages Of Growth ◽  
Rous Sarcoma ◽  
No Inhibition ◽  
Sarcoma Virus

Base layers of untransformed normal embryonic lung fibroblasts in different stages of growth were challenged with RSV (Rous sarcoma virus)-transformed bovine fibroblasts. The effect of this admixture on the growth of the RSV-transformed cells was studied. In all cases, the transformed cells proliferated freely, and it is concluded that the normal cells exerted no inhibition on the transformed cells.

scholarly journals Integration and expression of several molecular forms of Rous sarcoma virus DNA used for transfection of mouse cells.

1984 ◽  
Vol 4 (7) ◽  
pp. 1260-1269 ◽  
Author(s):  
P A Luciw ◽  
H Oppermann ◽  
J M Bishop ◽  
H E Varmus
Keyword(s):  
Cell Lines ◽  
Rous Sarcoma Virus ◽  
Linear Molecule ◽  
Viral Dna ◽  
Transformed Cell ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Sarcoma Virus ◽  
Mouse Cells ◽  
Transformed Cell Lines

To assess the factors required for integration and expression of retroviral DNA, we have examined viral DNA, RNA, and protein in NIH/3T3 mouse cells transformed by transfection with various forms of cloned Rous sarcoma virus (RSV) DNA. Linear RSV DNA molecules, derived from circular DNA containing two long terminal repeats (LTRs) and permuted by cleavage at the SacI restriction endonuclease site in the leader sequence, were integrated near the ends of the linear molecule, with the LTRs on the 3' side of the src gene. Integration of a subgenomic RSV DNA fragment containing the viral src gene without intact LTRs also occurred near the ends of the linear molecule. Head-to-tail tandem arrays of RSV DNA species were observed in some transformed cell lines that received fully digested DNA and in all cell lines that received DNA ligated to produce oligomers before transfection. Closed circular RSV DNA, with one or two LTRs, integrated without apparent specificity within several regions of the viral genome. After transfection with SacI-permuted RSV DNA still linked to arms of the lambda bacteriophage vector DNA, bacteriophage sequences were joined to host DNA. Transformed cell lines produced by transfection with the various forms of RSV DNA produced similar levels of viral src protein, although the efficiency of successful transformation varied by at least two orders of magnitude. Analyses of viral polyadenylated RNA, together with the patterns of viral DNA in transformed cells, indicated that viral DNA can be integrated and expressed without regard to LTR sequences, with adjacent host DNA presumably supplying signals required for the promotion and processing of functional src mRNA.

Integration and expression of several molecular forms of Rous sarcoma virus DNA used for transfection of mouse cells

1984 ◽  
Vol 4 (7) ◽  
pp. 1260-1269
Author(s):  
P A Luciw ◽  
H Oppermann ◽  
J M Bishop ◽  
H E Varmus
Keyword(s):  
Cell Lines ◽  
Rous Sarcoma Virus ◽  
Linear Molecule ◽  
Viral Dna ◽  
Transformed Cell ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Sarcoma Virus ◽  
Mouse Cells ◽  
Transformed Cell Lines

To assess the factors required for integration and expression of retroviral DNA, we have examined viral DNA, RNA, and protein in NIH/3T3 mouse cells transformed by transfection with various forms of cloned Rous sarcoma virus (RSV) DNA. Linear RSV DNA molecules, derived from circular DNA containing two long terminal repeats (LTRs) and permuted by cleavage at the SacI restriction endonuclease site in the leader sequence, were integrated near the ends of the linear molecule, with the LTRs on the 3' side of the src gene. Integration of a subgenomic RSV DNA fragment containing the viral src gene without intact LTRs also occurred near the ends of the linear molecule. Head-to-tail tandem arrays of RSV DNA species were observed in some transformed cell lines that received fully digested DNA and in all cell lines that received DNA ligated to produce oligomers before transfection. Closed circular RSV DNA, with one or two LTRs, integrated without apparent specificity within several regions of the viral genome. After transfection with SacI-permuted RSV DNA still linked to arms of the lambda bacteriophage vector DNA, bacteriophage sequences were joined to host DNA. Transformed cell lines produced by transfection with the various forms of RSV DNA produced similar levels of viral src protein, although the efficiency of successful transformation varied by at least two orders of magnitude. Analyses of viral polyadenylated RNA, together with the patterns of viral DNA in transformed cells, indicated that viral DNA can be integrated and expressed without regard to LTR sequences, with adjacent host DNA presumably supplying signals required for the promotion and processing of functional src mRNA.

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