scholarly journals Tumourigenicity, cell-surface glycoprotein changes and ornithine decarboxylase gene pattern in Ehrlich ascites-carcinoma cells

1985 ◽  
Vol 229 (3) ◽  
pp. 711-715 ◽  
Author(s):  
L Alhonen-Hongisto ◽  
A Kallio ◽  
R Sinervirta ◽  
O A Jänne ◽  
C G Gahmberg ◽  
...  
Keyword(s):  
Cell Surface ◽  
Cell Line ◽  
Ornithine Decarboxylase ◽  
Ehrlich Ascites Carcinoma ◽  
Tumour Cells ◽  
Carcinoma Cells ◽  
Surface Glycoprotein ◽  
Cell Surface Glycoprotein ◽  
Phenotypic Changes ◽  
Ehrlich Ascites

We selected a 2-difluoromethylornithine-resistant Ehrlich ascites-carcinoma cell line that grows in the presence of 20 mM-difluoromethylornithine. These cells contain 10-20 times the normal amount of hybridizable sequences for ornithine decarboxylase (EC 4.1.1.17) in their genomic DNA. We used these gene-amplified cells, their revertant counterparts (grown in the absence of the drug after an established gene amplification) and tumour cells grown in the presence of putrescine to investigate the changes of ornithine decarboxylase gene pattern and simultaneously occurring phenotypic changes, such as tumourigenicity and the expression of cell-surface glycoproteins. In the tumour cells reverted back to the normal gene frequency, not only did the amplified sequences disappear, but there were also signs of gene re-arrangements seen as a ‘gene jump’, when a signal evidently moved to a heavier restriction fragment. Similar gene re-arrangement likewise occurred in cells exposed to putrescine. Although the wild-type tumour cells and the gene-amplified cells readily grew in the peritoneal cavity of mice, the revertant cells and the putrescine-treated cells had lost their tumourigenicity in mice. Gene-amplified tumour cells and the revertant cells showed distinct changes in their surface glycoprotein pattern in comparison with the parental cell line. These findings indicate that alterations of ornithine decarboxylase gene pattern/dosage may be associated with phenotypic changes possibly related to the tumourigenicity of these carcinoma cells.

scholarly journals Mouse and human ornithine decarboxylase genes. Methylation polymorphism and amplification

1987 ◽  
Vol 242 (1) ◽  
pp. 205-210 ◽  
Author(s):  
L Alhonen-Hongisto ◽  
P Leinonen ◽  
R Sinervirta ◽  
R Laine ◽  
R Winqvist ◽  
...  
Keyword(s):  
Cell Line ◽  
Ornithine Decarboxylase ◽  
Ehrlich Ascites Carcinoma ◽  
Myeloma Cell ◽  
Restriction Pattern ◽  
Restriction Endonucleases ◽  
Cell Leukaemia ◽  
Chromosome 2 ◽  
Ehrlich Ascites ◽  
Mouse Leukaemia

With the use of the isoschizomeric restriction endonucleases HpaII and MspI, we found that mouse tumour ornithine decarboxylase (ODC; EC 4.1.1.17) genes are extensively methylated. ODC genes in L1210 mouse leukaemia cells were apparently more methylated than in Ehrlich ascites carcinoma, as revealed by the use of HpaII endonuclease, yet the digestion of genomic DNA isolated from these two murine tumour cell lines with MspI, which cleaves at a CCGG sequence, also with internally methylated cytosine, resulted in an apparently identical restriction pattern. It is possible that the amplification of ODC genes in Ehrlich ascites-carcinoma cells in response to 2-difluoromethylornithine (DFMO) was associated with hypomethylation, or that less-methylated genes were amplified. A human myeloma (Sultan) cell line only revealed three separate hybridization signals when cleaved with HpaII. One of these signals was amplified under the pressure of DFMO. When cleaved with MspI, these three HpaII fragments disappeared and were replaced by a double signal of 2.3-2.4 kilobase-pairs (kbp) in size. The amplified ODC sequences in the Sultan myeloma cell line apparently originated from chromosome 2, as indicated by a unique hybridization signal in a 5.8 kbp HindIII fragment specific for the human ODC locus on chromosome 2. A comparison of different human cells, the Sultan myeloma, a lymphocytic B-cell leukaemia (Ball), normal mononuclear leucocytes and leucocytes obtained from leukaemia patients, revealed interesting differences in the methylation of ODC genes. The use of two restriction endonucleases (HpaII and CfoI), the cleavage site for both of which contains a CG sequence and which only cleave when cytosine is unmethylated, indicated that ODC genes in the lymphocytic leukaemia cells were much less methylated than those in the normal leucocytes or in the Sultan cells.

scholarly journals Polyamines in mycoplasmas and in mycoplasma-infected tumour cells

1982 ◽  
Vol 202 (1) ◽  
pp. 267-270 ◽  
Author(s):  
L Alhonen-Hongisto ◽  
P Veijalainen ◽  
C Ek-Kommonen ◽  
J Jänne
Keyword(s):  
Ornithine Decarboxylase ◽  
Ehrlich Ascites Carcinoma ◽  
Carcinoma Cells ◽  
Leukemia Cells ◽  
Animal Cells ◽  
Irreversible Inhibitor ◽  
Ehrlich Ascites ◽  
Infected Cells ◽  
L1210 Mouse Leukemia ◽  
High Concentrations

Three out of four different mycoplasma strains analysed for the polyamine contents contained relatively high concentrations of putrescine, cadaverine, spermidine and spermine. In addition to ornithine decarboxylase (EC 4.1.1.17) activity, the mycoplasmas also exhibited comparable or higher lysine decarboxylase (EC 4.1.1.18) activity fully resistant to the action of 2-difluoromethylornithine, an irreversible inhibitor of eukaryotic ornithine decarboxylase. 2-Difluoromethylornithine did not modify the polyamine pattern of actively growing mycoplasmas. Ehrlich ascites carcinoma cells and L1210 mouse leukemia cells infected with any of the four mycoplasma strains contained, in addition to putrescine, spermidine and spermine, and also easily measurable concentrations of cadaverine; the latter diamine was absent in uninfected cultures. When the infected cells were exposed to difluoromethylornithine, the accumulation of cadaverine was markedly enhanced. The modification of cellular polyamine pattern by mycoplasmas, especially in the presence of inhibitors of eukaryotic ornithine decarboxylase, could conceivably be used as an indicator of mycoplasma infection in cultured animal cells.

scholarly journals Molecular characterization and expression of a novel human leukocyte cell-surface marker homologous to mouse Ly-9

Blood ◽  
2001 ◽  
Vol 97 (11) ◽  
pp. 3513-3520 ◽  
Author(s):  
Miguel Angel de la Fuente ◽  
Victoria Tovar ◽  
Neus Villamor ◽  
Nuria Zapater ◽  
Pilar Pizcueta ◽  
...  
Keyword(s):  
Cell Surface ◽  
B Cell ◽  
Cell Line ◽  
B Lymphocytes ◽  
Full Length ◽  
Surface Glycoprotein ◽  
T And B Lymphocytes ◽  
Cdna Clones ◽  
Cell Surface Glycoprotein ◽  
Full Length Cdna

Ly-9 is a mouse cell-surface glycoprotein that is selectively expressed on thymocytes and on mature T and B lymphocytes. Ly-9 belongs to the CD2 subset of the immunoglobulin superfamily, an emerging family of cell signaling receptors. Recently, a partial human Ly-9 complementary DNA (cDNA) sequence has been described. Full-length cDNA clones were isolated that included the initiation codon, the sequence encoding the full signal peptide, and 14 amino acids more in the cytoplasmic domain than in the previously reported clone. The predicted extracellular domain of human Ly-9 contains 4 immunoglobulinlike domains, similar to those in mouse Ly-9. Northern blot analysis revealed that the human Ly-9 messenger RNA (2.6 kb) is expressed predominantly in lymph node, spleen, thymus, and peripheral blood leukocytes. Four monoclonal antibodies (mAbs) were raised against human Ly-9 by immunizing mice with the pre-B-cell line 300.19 stably transfected with human Ly-9 full-length cDNA. These mAbs strongly stained the surfaces of cells transfected with human Ly-9 cDNA but not of untransfected cells. Human Ly-9 expression was restricted to T and B lymphocytes and thymocytes, with the highest levels of expression on CD4+CD8− and CD4−CD8+ thymocytes. Monocytes, granulocytes, platelets, and red blood cells were uniformly negative for Ly-9. These mAbs immunoprecipitated major polypeptides of 120 kd from the transfected cells and 120 kd and 100 kd from B-cell line Daudi, probably because of the cell-surface–expressed isoforms. These data demonstrate that human Ly-9 is a new marker for the study of normal and malignant leukocytes.

Difluoromethylornithine-induced amplification of ornithine decarboxylase genes in Ehrlich ascites carcinoma cells

1985 ◽  
Vol 126 (2) ◽  
pp. 734-740 ◽  
Author(s):  
L. Alhonen-Hongisto ◽  
A. Kallio ◽  
R. Sinervirta ◽  
P/ Seppänen ◽  
K.K. Kontula ◽  
...  
Keyword(s):  
Ornithine Decarboxylase ◽  
Ehrlich Ascites Carcinoma ◽  
Carcinoma Cells ◽  
Ehrlich Ascites

scholarly journals Deficiency of lymphocyte function-associated antigen 3 (LFA-3) in paroxysmal nocturnal hemoglobinuria. Functional correlates and evidence for a phosphatidylinositol membrane anchor.

1987 ◽  
Vol 166 (4) ◽  
pp. 1011-1025 ◽  
Author(s):  
P Selvaraj ◽  
M L Dustin ◽  
R Silber ◽  
M G Low ◽  
T A Springer
Keyword(s):  
Cell Membrane ◽  
Cell Surface ◽  
Cell Line ◽  
Paroxysmal Nocturnal Hemoglobinuria ◽  
Lymphoma Cell ◽  
Surface Glycoprotein ◽  
Thymic Epithelial Cells ◽  
Lymphocyte Function ◽  
Lymphoma Cell Line ◽  
Cell Surface Glycoprotein

Lymphocyte function-associated antigen 3 (LFA-3) is a widely distributed cell surface glycoprotein that binds to the T lymphocyte CD2 surface glycoprotein. This interaction mediates CTL-target cell conjugate formation and adhesion of thymocytes to thymic epithelial cells. CD2 is also the E rosette receptor of T lymphocytes and mediates rosetting with autologous E by binding to LFA-3. We describe deficient expression of LFA-3 on E from paroxysmal nocturnal hemoglobinuria (PNH) patients. PNH is an acquired defect affecting phosphatidylinositol-anchored membrane proteins, of which decay-accelerating factor (DAF) is most important in the clinical symptoms of PNH. LFA-3-negative, weakly positive, and positive populations were found among PNH E. There was a good correlation with DAF deficiency. PNH E exhibited decreased binding of 125I-CD2 and rosetting with a human T lymphoma cell line. PNH E readily incorporated purified LFA-3, restoring LFA-3 expression and the CD2 binding and rosetting activity to normal levels. The expression of DAF was not restored after the incorporation of purified LFA-3 into PNH E, showing that LFA-3 and DAF are different molecules. Phosphatidylinositol-specific phospholipase C (PIPLC) treatment of a B lymphoma cell line released 35% of the cell surface LFA-3 and 62% of DAF. LFA-3 on E was resistant to PIPLC. However, when LFA-3 purified from human E was reconstituted in sheep E or human E and subjected to PIPLC treatment, 40-50% of LFA-3 was released from the cell membrane. The results show that LFA-3 is attached to the cell membrane by a phosphatidylinositol glycolipid moiety, and confirm previous findings (37-41) that LFA-3 is a cell adhesion molecule that mediates adhesion by interacting with CD2 antigen.

scholarly journals Transfer of intestine-derived diamines into tumour cells during treatment of Ehrlich-ascites—carcinoma-bearing mice with polyamine anti-metabolites

1984 ◽  
Vol 218 (2) ◽  
pp. 641-644 ◽  
Author(s):  
A Kallio ◽  
P Nikula ◽  
J Jänne
Keyword(s):  
General Circulation ◽  
Diamine Oxidase ◽  
Ehrlich Ascites Carcinoma ◽  
Tumour Cells ◽  
Carcinoma Cells ◽  
Ehrlich Ascites

Treatment of Ehrlich-ascites-carcinoma-bearing mice with methylglyoxal bis(guanylhydrazone) alone or in combination with 2-difluoromethylornithine greatly enhanced the transfer of intragastrically administered radioactive putrescine and cadaverine into the carcinoma cells. Difluoromethylornithine alone did not have any effect on the accumulation of intestine-derived diamines in the tumour cells. The frequently reported restoration of difluoromethylornithine-induced polyamine depletion on administration of methylglyoxal bis(guanylhydrazone) is in all likelihood attributable to a profound inhibition of intestinal diamine oxidase (EC 1.4.3.6), resulting in an enhanced entry of intestinal (bacterial) diamines into general circulation and finally into tumour cells.

scholarly journals Intracellular putrescine and spermidine deprivation induces increased uptake of the natural polyamines and methylglyoxal bis(guanylhydrazone)

1980 ◽  
Vol 192 (3) ◽  
pp. 941-945 ◽  
Author(s):  
L Alhonen-Hongisto ◽  
P Seppänen ◽  
J Jänne
Keyword(s):  
Transport System ◽  
Culture Medium ◽  
Ehrlich Ascites Carcinoma ◽  
Tumour Cells ◽  
Carcinoma Cells ◽  
Polyamine Synthesis ◽  
Ascites Tumour ◽  
Proliferative Effect ◽  
Ehrlich Ascites ◽  
Ascites Tumour Cells

Inhibition of polyamine synthesis by alpha-difluoromethylornithine in cultured Ehrlich ascites-carcinoma cells rapidly enhanced the uptake of exogenous putrescine, spermidine and spermine from the culture medium. In tumour cells exposed to the drug for 2 days, the intracellular concentration of spermidine was decreased to less than 10% of that found in untreated cells. However, the strikingly stimulated transport system brought the concentration of spermidine to the control values in less than 2h after supplementation of the cells with micromolar concentrations of the polyamine. In the absence of polyamine deprivation, tumour cells did not accumulate extracellular polyamines to any appreciable extent. Ascites-tumour cells deprived of putrescine and spermidine likewise concentrated methylglyoxal bis(guanylhydrazone) [1,1′-[methylethanedylidine)dinitrilo]diguanidine] at a greatly enhanced rate. A previous “priming of tumour cells with difluoromethylornithine followed by an exposure of the cells to methylglyoxal bis(guanylhydrazone) resulted in a marked and rapid anti-proliferative effect.

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