- Primary research
- Open Access
Effect of 5-azacytidine and galectin-1 on growth and differentiation of the human b lymphoma cell line bl36
© Poirier et al; licensee BioMed Central Ltd. 2001
Received: 7 August 2001
Accepted: 17 December 2001
Published: 17 December 2001
5-AzaCytidine (AzaC) is a DNA demethylating drugs that has been shown to inhibit cell growth and to induce apoptosis in certain cancer cells. Induced expression of the galectin1 (Gal1) protein, a galactoside-binding protein distributed widely in immune cells, has been described in cultured hepatoma-derived cells treated with AzaC and this event may have a role in the effect of the drug. According to this hypothesis, we investigated the effect of AzaC and Gal1 on human lymphoid B cells phenotype.
The effect of AzaC and Gal1 on cell growth and phenotype was determined on the Burkitt lymphoma cell line BL36. An immunocytochemical analysis for detection of Gal1 protein expression was performed in AzaC-treated cells. To investigate the direct effects of Gal1, recombinant Gal1 was added to cells.
Treatment of lymphoid B cells with AzaC results in: i) a decrease in cell growth with an arrest of the cell cycle at G0/G1 phase, ii) phenotypic changes consistent with a differentiated phenotype, and iii) the expression of p16, a tumor-suppressor gene whose expression was dependent of its promoter demethylation, and of Gal1. A targeting of Gal 1 to the plasma membrane follows its cytosolic expression. To determine which of the effects of AzaC might be secondary to the induction of Gal1, recombinant Gal1 was added to BL36 cells. Treated cells displayed growth inhibition and phenotypic changes consistent with a commitment toward differentiation.
Altered cell growth and expression of the cell surface plasma cell antigen, CD138 are detectable in BL36 cells treated by AzaC as well as by Gal1. It seems that AzaC-induced Gal1 expression and consequent binding of Gal1 on its cell membrane receptor may be, in part, involved in AzaC-induced plasmacytic differentiation.
DNA methylation is involved in cellular development, differentiation and transformation . In different types of tumours, aberrant methylation of CpG islands in the promoter region has been observed for many differentiation- and cancer-related genes resulting in the silencing of their expression . Therefore, over the past decade, there has been increasing interest in the use of demethylating agents to induce the differentiation or the apoptosis of cancer cells [3, 4]. Treatment of the cells with the pyrimidine analogue 5-AzaCytidine (AzaC), which inhibits methylation of cytosine residues during replication in the newly synthesised DNA, has been demonstrated to reactivate the expression of many silenced genes, as well as the expression of the silenced retro viral genomes . Silencing of one of the most important cell cycle regulatory proteins p16INK4a by methylation of the CpG islands in the promoter region has been found to be a common event in tumours [6, 7]. Protein p16 suppresses S-phase entry by antagonising the cyclin-dependent kinases CDK4 and CDK6 .
Deciphering the molecular mechanisms underlying the phenotypic effects of the treatment with demethylating drugs is a crucial step in understanding what genes may be interesting targets for chemotherapy. The available data on the mechanism of action of these drugs strengthen the idea that it is different from that of agents that act primarily via their cytotoxic effects, such as Arc-C . Several lines of evidence suggest that galectin-1 (Gal-1), a 14 kDa galactoside-binding protein distributed widely in immune cells, could be involved in these mechanisms. Several members of the galectin family have been found to modulate cell differentiation and cell survival [10–15]. Early studies demonstrated that the expression of Gal1 could be induced in cultured hepatoma-derived cells by treatment with AzaC . Chiariotti and co-workers showed that reactivation of the silent Gal1 alleles is accompanied by a transition from a fully methylated to a fully unmethylated state of several CpG dinucleotides in the promoter region . In addition, nonexpressing tissues exhibited highly heterogeneous methylation profiles . Gal is considered to be a typical cytosolic protein, lacking a signal peptide for membrane translocation . However, most of the functions assigned to galectins are confined to the cell surface or extracellular milieu [10, 20, 21], consistent with evidences for extracellular roles of Gal1 in regulation of cellular differentiation and proliferation. It is clear that Gal1 can be specifically secreted and targeted by an infrequent mechanism [22–24]. The constitutive expression as well as the secretion of Gal1 dramatically depend on cell types  and are responsive to developmental events [20, 22, 23].
An example is found during erythroid differentiation of the K562 human leukaemia cell line. During differentiation induced by erythropoietin and deprivation of granulocyte-macrophage colony-stimulating factor, the cells empty their cytoplasmic content of endogenous Gal1 into the external medium where it is bind to cell surface receptors . The synthesis and secretion of Gal1 by leukocytes are of interest because lactosaminoglycans present at the leukocyte cell surface may be physiologically significant galectin receptors that could mediate autocrine or paracrine functions. Several lines of evidence indicate that Gal1 may function as an autocrine negative growth regulator or as a pro-apoptotic factor [26–28]. We have recently demonstrated that Gal1 binding to Burkitt lymphoma cells results in an intracellular signal, with inhibition of the tyrosine phosphatase activity of CD45 and therefore phosphorylation of Lyn kinase [29, 30].
In this work, we study the effect of AzaC treatment on the lymphoma cell line BL36. As p16INK4a gene has been found to be downregulated by hypermethylation at high frequency in different types of tumours [6, 7], it is used as a control of AzaC effect. Then, the phenotypic effects of AzaC are compared to those obtained by the addition of exogenous Gal1. The findings that we report here lend further support to a potential role for Gal1 in the AzaC-induced pathway of differentiation in hematopoietic cells.
Materials and Methods
The BL36 B lymphoma cell line , a gift from Pr. Lenoir (CIRC, Lyon, France) to Pr. M. Raphael, was maintained in a complete medium of RPMI 1640, containing 10% heat-inactivated foetal calf serum, 2 mM L-Glutamine, 1 mM sodium pyruvate (complete medium). Treatment with AzaC was conducted as follows: 24 hours after seeding 5, 10 or 50 μM of AzaC were added to 5 ml of complete medium [16, 32]. The cells were feeded with the appropriate medium during the time of the experiments.
Several independent methods were used to assess the proliferative versus death states of the cells. For direct determination of cell number, at the indicated time point the cells were harvested and counted using a Coulter Counter (Beckman Coulter France, Villepinte, France). Determination of the viable cell numbers used propidium iodide (PI) (Sigma, Saint-Quentin Fallavier, France). To determine the percent cell death including both apoptotic and necrotic cells, AzaC-treated cells were analyzed for AnnexinV binding and PI uptake as described  using the ApoDetect AnnexinV-FITC kit (Zymed). Briefly, after washing of cultured cells (3.105) with PBS and resuspension in binding buffer, the cells were stained with 10 μl FITC-labelled AnnexinV and 10 μl PI (20 μg/ml). After 5 min of incubation at room temperature in the dark, again 200 μl of binding buffer was added and cells were analyzed by flow cytometry. The percent cell death was calculated by determining the percent of viable cells:
The MTS assay (CellTiter 96™, Promega, Charbonnières, France)  was used to compare the percentage of metabolically active cells in treated cells vs. untreated cells, as previously described . To study the cell cycle, 2.106 BL36 cells were pelleted at 1000 g for 5 minutes and the pellet was incubated in a mixture of ethanol-PBS (70/30 v/v). The cells were pelleted another time and suspended in 800 μl PBS. One hundred microliters of 1 mg/ml RNase was added to the cell suspension. One hundred microliters of 400 μg/ml PI was added to the solution to stain the nuclear DNA. The DNA content of the cells was determined by a flow cytometer, FACScan (Becton Dickinson, Mountain View, CA), and the percentages of cells in G0+G1, S, and G2+M phases of the cell cycle were analyzed by a polynomial model (SFIT, Becton Dickinson).
To determine the cell phenotypes, cells were suspended in PBS supplemented with 2% BSA, and incubated in suspension for 30 min with the fluorescently tagged primary antibody or negative control, or incubated with untagged antibody, washed, and incubated again for 30 min with FITC-conjugated second antibody. All incubations were performed at 4°C. Flow cytometry was performed using a Coulter Epics Elite ESP instrument.
Human recombinant Gal1 (rGal1) was obtained as described elsewhere . The protein was purified by affinity chromatography on a lactosyl-divinylsulfone-agarose column. Antibodies against human Gal1 were generated as described in  and . The immunolocalization of Gal1 was carried out as described for K562 cells .
For the preparation of cell extracts, a total of 17.106 cells were solubilized in 1 ml of extraction buffer: 50 mM Tris pH 7.4, 150 mM NaCl, 5 mM EDTA, 0.1% SDS (w/v), 0.5% sodium deoxycholate (w/v), 0.5% Nonidet NP40 (v/v). Finally, 1 tablet of antiprotease cocktail (Roche, Meylan, France) was added to 10 ml of buffer. The cells were sonicated in ice three times. The lysat was centrifuged at 100, 000 g for 15 min at 4°C. The supernatant was collected and stored at -20°C until used. Proteins were resolved by discontinuous SDS-PAGE on 1.5 mm gel (T: 6–18%) according to the method of Laemmli. . For Western blotting, proteins separated by SDS-PAGE were electrotransferred onto Immobilon-P membrane (Millipore). Blots were incubated with 1:50 anti-Gal1 mAb, or 1:500 anti-pl6 mAb (Pharmingen) for 2 h at room temperature. The blots were developed with anti-mouse Ig antibody-HRP diluted 1/10,000, followed by incubation in the Amplified Opti-4CN kit substrate (Bio-Rad). The image of the membranes was acquired from GS-700 Densitometer 4, and analyzed with Molecular Analyst Software (Bio-Rad).
Modulation of urface Markers on BL36 Cells by AzaC
Treatment AzaC = 5 μM
AzaC- 10 μM
AzaC = 50 μM
CD 45 RA
CD 45 RO
Modulation of Surface Markers on BL36 Cells by rGal1
Control 24 h
Treatment (% positive) rGal1 – 24 h
Control 48 h
Treatment (% positive) rGal1-48 h
In 1994, Chiariotti et al.  first reported experimental data showing that the expression of Gal1 can be induced in cultured hepatoma-derived cells by treatment with AzaC. Interestingly, Gal1 and AzaC individually have been shown to affect similar cell processes in cancer cells, including differentiation, and growth inhibition [12–15, 27, 29, 46–51]. Available data are consistent with the suggestion that the expression of Gal1, accompanied by its secretion and its binding to cell surface receptors, could be involved in the AzaC observed effects in hematopoietic cells where Gal1 modulates differentiation or apoptosis. However, the mechanisms of these effects on hematopoietic cells are unclear yet.
In the present study, the effects of AzaC, on the cell phenotype, cell differentiation and cell death of BL36 cells were analyzed. The effects on cellular growth were time- and dose-dependent. Five μM AzaC caused significant inhibition on cellular growth, but the cell viability remained practically unchanged. AzaC increased the cell fraction in the G0/G1 phases, suggesting that AzaC inhibits cell division, which may be one of critical mechanisms of cell modulation by AzaC. AzaC-treated cells initially may become arrested at the G1 phase and then may either escape to the cycle arrest or die due to mechanisms leading to programmed cell death.
We also show that incubation of BL36 with AzaC induces expression of Gal1. AzaC is thought to exert its effects as a competitive inhibitor of cytosine methylation, resulting in the expression of silenced genes. The gene for Gal1 is one whose expression is possibly enhanced in this manner. In the results reported here, we found that Gal 1 was detected in cytosol after 120 h of treatment by 10 μM AzaC and then Gal1 was externalized and bound to cell surface receptors 24 h later. A key to understanding the extracellular biological functions of Gal1 is how its secretion appears to be regulated and re-directed during development and differentiation. Gal1 is likely released form vesicles close to the plasma membrane. On the basis of the data, we propose that the released Gal1 be immediately recruited to modulate cell activity. Gal1 may do this by interacting with and modulating cell receptors via its carbohydrate recognition domains because the Gal1-receptor interaction is abrogated by thiodigalactoside .
Others and we have previously reported that Gal1 binds to T and B lymphoblastoid cells [29, 52, 53]. Other studies have demonstrated that galectins are immunosuppressive, in animal models of autoimmune diseases [54–56]. Whereas the full role of Gal1 in modulating immune function is not yet understood, the increase in Gal1 expression by AzaC in BL cells suggests that Gal1 may play a role in the behaviour of normal leukocytes and of tumour cells.
What is the underlying mechanism? Although the regulatory machinery triggered by demethylating stimulus and resulting in phenotype modifications is not yet elucidated, it probably involves the stimulation of a signalling cascade that regulates cell proliferation and viability. A recently proposed model for such a cascade suggests the involvement of a cytoplasmic protein, AZ2 . The amino-terminal part of the AZ2 protein is homologous to the previously reported TANK and I-TRAF, which participate in the signal transduction cascade from the TNF-receptor to the transcription factor NFkappaB. Demethylating stimulus may also modify a pathway activated by the membrane-anchored protein-tyro sine phosphatase CD45. Engagement of CD45 is known to regulate Src tyrosine kinases phosphorylation, phospholipase Cγ regulation, inositol phosphate production, diacylglycerol production, PKC activation, and calcium mobilisation [58–60].
Increased synthesis and secretion of Gal1 by the cell could account for part of the phenotypic alterations detected in AzaC treated cells. Gal1-induced dimerisation and/or segregation might inhibit the catalytic site in CD45, thereby blocking tyrosine phosphates activity. Because Gal1 binding to cell surface receptors results in tyrosine phosphorylation [29, 61], it may allow a kinase-dependent signal to be transduced. Several studies have linked Gal1 expression with growth inhibition  and cell death [26, 62]. However, the reports that some growth inhibitory agents did not induce Gal1 expression indicated that Gal1 expression is not dependent on the cell's growth state in general, through it may be involved in growth suppression . Moreover, it is likely that Gal1 acts in a manner to regulate specific signal transduction processes that is determined by the cell type and by the state of cell differentiation. In this work, exogenous rGal1 added to BL cells inhibited cell growth. Moreover, Gal1 as well as AzaC induced an expression of the cell surface plasma cell antigen, CD138, a phenotypic marker that identify cells with plasmacytic differentiation . This is consistent with the hypothesis that AzaC and Gal1 share similar signals for differentiation, however, since there was a significant difference in the expression of CD19 and CD23 after AzaC or Gal1 treatments it is likely that some pathways are specifically modified by AzaC. The mechanisms involved in these different pathways, important in clinical therapy, remain to be elucidated in the future. Ongoing studies are aimed at identifying as globally as possible the modifications resulting from AzaC treatment by using proteomics .
This work was supported, in part, by grants from the Ministère de l'Education Nationale de la Recherche et de la Technologie (MENRT), and from the Ligue Française contre le Cancer (Comité de Seine Saint-Denis). FP was supported by ARC (Association pour la Recherche sur le Cancer).
- Schmutte C, Jones PA: Involvement of DNA methylation in human carcinogenesis. Biol Chem. 1998, 379: 377-388.View ArticlePubMedGoogle Scholar
- Momparler RL, Bovenzi V: DNA methylation and cancer. J Cell Physiol. 2000, 183: 145-154. 10.1002/(SICI)1097-4652(200005)183:2<145::AID-JCP1>3.0.CO;2-V.View ArticlePubMedGoogle Scholar
- Murakami T, Li X, Gong J, Bhatiah U, Traganos F, Darzynkiewicz Z: Induction of apoptosis by 5-azacytidine: drug concentration-dependent differences in cell cycle specificity. Cancer Res. 1995, 55: 3093-3098.PubMedGoogle Scholar
- Wang XM, Wang X, Li J, Evers BM: Effects of 5-azacytidine and butyrate on differentiation and apoptosis of hepatic cancer cell lines. Ann. Surg. 1998, 227: 922-931. 10.1097/00000658-199806000-00016.PubMed CentralView ArticlePubMedGoogle Scholar
- Masucci MG, Contreras-Salazar B, Ragnar E, Falk K, Minarovits J, Ernberd I, Klein G: 5-Azacytidine up regulates the expression of EBV nuclear antigen 2 (EBNA2) through ENBNA6 and latent membrane protein in Burkitt's lymphoma line rael. J. Virol. 1989, 63: 3135-3141.PubMed CentralPubMedGoogle Scholar
- Okamoto A, Demetrick DJ, Spillare EA, Hagiwara K, Hussain SP, Bennett WP, Forrester K, Gerwin B, Serrano M, Beach DH: Mutations and altered expression of pl6INK4 in human cancer. Proc Natl Acad Sci USA. 1994, 91: 11045-11049.PubMed CentralView ArticlePubMedGoogle Scholar
- Merlo A, Herman JG, Mao L, Lee DJ, Gabrielson E, Burger PC, Baylin SB, Sidransky D: CpG island methylation is associated with transcriptional silencing of the tumour suppressor pl6/CDKN2/MTS1 in human cancers. Nat Med. 1995, 1: 686-692.View ArticlePubMedGoogle Scholar
- Singal R, Ginder GD: DNA methylation. Blood. 1999, 93: 4059-4070.PubMedGoogle Scholar
- Lubbert M: DNA methylation inhibitors in the treatment of leukemias, myelodysplastic syndroms, hemoglobinopathies: clinical results and possible mechanisms of action. Curr. Top. Microbiol. Immunol. 2000, 249: 135-164.PubMedGoogle Scholar
- Hadari YR, Arbel-Goren R, Levy Y, Amsterdam A, Alon R, Zakut R, Zick Y: Galectin-8 binding to integrins inhibits cell adhesion and induces apoptosis. J Cell Sci. 2000, 113: 2385-2397.PubMedGoogle Scholar
- Adams L, Scott GK, Weinberg C: Biphasic modulation of cell growth by recombinant human galectin-1. Biochim. Biophys. Acta,. 1996, 1312: 137-144. 10.1016/0167-4889(96)00031-6.View ArticleGoogle Scholar
- Allione A, Wells V, Forni G, Mallucci L, Novelli F: Beta-galactoside-binding protein (beta GBP) alters the cell cycle, up-regulates expression of the alpha- and beta-chains of the IFN-gamma receptor, and triggers IFN-gamma-mediated apoptosis of activated human T lymphocytes. J Immunol. 1998, 161: 2114-2119.PubMedGoogle Scholar
- Blaser C, Kaufmann M, Muller C, Zimmermann C, Wells V, Mallucci L, Pircher H: Beta-galactoside-binding protein secreted by activated T cells inhibits antigen-induced proliferation of T cells. Eur. J. Immunol. 1998, 28: 2311-2319. 10.1002/(SICI)1521-4141(199808)28:08<2311::AID-IMMU2311>3.0.CO;2-G.View ArticlePubMedGoogle Scholar
- Fouillit M, Lévi-Strauss M, Giudicelli V, Lutomski D, Bladier D, Caron M, Joubert-Caron R: Affinity purification and characterization of recombinant human galectin-1. J. Chromatogr. 1998, 706: 167-171. 10.1016/S0378-4347(97)00336-8.View ArticleGoogle Scholar
- Perillo NL, Marcus ME, Baum LG: Galectins: versatile modulators of cell adhesion, cell proliferation, and cell death. J. Mol. Med. 1998, 76: 402-412. 10.1007/s001090050232.View ArticlePubMedGoogle Scholar
- Chiariotti L, Benvenuto G, Zarrilli R, Rossi E, Salvatore P, Colantuoni V, Bruni CB: Activation of the galectin-1 (L-14-I) gene from nonexpressing differentiated cells by fusion with undifferentiated and tumorigenic cells. Cell Growth Differ. 1994, 5: 769-775.PubMedGoogle Scholar
- Benevenuto G, Carpentieri M, Salvatore P, Cindolo L, Bruni CB, Chiariotti L: Cell-specific transcriptional regulation and reactivation of Galectin-1 gene expression are controlled by DNA methylation of the promoter region. Mol. and Cell. Biol. 1996, 16: 2736-2743.View ArticleGoogle Scholar
- Salvatore P, Benvenuto G, Caporaso M, Bruni CB, Chiariotti L: High resolution methylation analysis of the galectin-1 gene promoter region in expressing and nonexpressing tissues. FEBS Lett. 1998, 421: 152-158. 10.1016/S0014-5793(97)01553-6.View ArticlePubMedGoogle Scholar
- Bladier D, Le Caër J-P, Joubert R, Caron M, Rossier J: β-galactoside soluble lectin from human brain: a complete amino acid sequence. Neurochem. Int. 1991, 18: 275-281. 10.1016/0197-0186(91)90195-J.View ArticlePubMedGoogle Scholar
- Hughes RC: Secretion of the galectin family of mammalian carbohydrate-binding proteins. Biochim Biophys Acta. 1999, 1473: 172-185. 10.1016/S0304-4165(99)00177-4.View ArticlePubMedGoogle Scholar
- Mehul B, Hughes R: Plasma membrane targetting, vesicular budding and release of galectin 3 from the cytoplasm of mammalian cells during secretion. J. Cell Sci. 1997, 110: 1169-1178.PubMedGoogle Scholar
- Cooper DNW, Barondes SH: Evidence for export of a muscle lectin from cytosol to extracellular matrix and for a novel secretory mechanism. J. Cell Biol. 1990, 110: 1681-1691.View ArticlePubMedGoogle Scholar
- Avellana-Adalid V, Rebel G, Caron M, Cornillot JD, Bladier D, Joubert-Caron R: Changes in S-type lectin localization in neuroblastoma cells (N1E115) upon differentiation. Glycoconj. J. 1994, 11: 286-291.View ArticlePubMedGoogle Scholar
- Lutomski D, Fouillit M, Bourin P, Mellottée D, Denize N, Pontet M, Bladier D, Caron M, Joubert-Caron R: Externalization and binding of galectin-1 on cell surface of K562 cells upon erythroid differentiation. Glycobiology. 1997, 7: 1193-1199.View ArticlePubMedGoogle Scholar
- Lutomski D, Denize N, Mellottée D, Bourin P, Pontet M, Bladier D, Caron M, Joubert-Caron R: Differential expression of a β-galactoside binding lectin (galectin 1) in human erythroleukemia cell lines: TF1 and K562. Blood. 1996, 88: 118b-Google Scholar
- Goldstone SD, Lavin MF: Isolation of a cDNA clone, encoding a human β-galactoside binding protein, overexpressed during glucocorticoid-induced cell death. Biochem. Biophys. Res. Comm. 1991, 178: 746-750.View ArticlePubMedGoogle Scholar
- Wells V, Mallucci L: Identification of an autocrine negative growth factor: mouse β-galactoside-binding protein is a cytostatic factor and cell growth regulator. Cell. 1991, 64: 91-97.View ArticlePubMedGoogle Scholar
- Allione A, Bernabei P, Rigamonti L, Bertolaccini L, Mallucci L, Forni G, Novelli F: Differential IFNγR expression controls the growth or apoptosis of human malignant T cells. Fund. din. Immunol. 1995, 3: 66-Google Scholar
- Fouillit M, Joubert-Caron R, Poirier F, Bourin P, Monostori E, Levi-Strauss M, Raphael M, Bladier D, Caron M: Regulation of CD45-induced signaling by galectin-1 in Burkitt lymphoma B cells. Glycobiology. 2000, 10: 413-419. 10.1093/glycob/10.4.413.View ArticlePubMedGoogle Scholar
- Fouillit M, Poirier F, Monostori E, Raphael M, Bladier D, Joubert-Caron R, Caron M: Analysis of galectin 1-mediated cell signaling by combined precipitation and electrophoresis techniques. Electrophoresis. 2000, 21: 275-280. 10.1002/(SICI)1522-2683(20000101)21:2<275::AID-ELPS275>3.3.CO;2-0.View ArticlePubMedGoogle Scholar
- Favrot MC, Maritaz O, Suzuki T, Cooper M, Philip I, Philip T, Lenoir G: EBV-negative and -positive Burkitt cell lines variably express receptors for B-cell activation and differentiation. Int J Cancer. 1986, 38: 901-906.View ArticlePubMedGoogle Scholar
- Cuomo L, Triverdi P, de Campos-Lima PO, Zhang QJ, Ragnar E, Klein G, Masucci MG: Selective induction of allostimulatory capacity after 5-azaC treatment of EBV carrying but not EBV negative Burkitt lymphoma cell lines. Mol. Immunol. 1993, 30: 441-450. 10.1016/0161-5890(93)90112-O.View ArticlePubMedGoogle Scholar
- Koopman G, Reutelingsperger CP, Kuijten GA, Keehnen RM, Pals ST, van Oers MH: Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptosis. Blood. 1994, 84: 1415-1420.PubMedGoogle Scholar
- Buttke TM, McCubrey JA, Owen TC: Use of an aqueous soluble tetrazolium:formazan assay to measure viability and proliferation of lymphokine-dependent cell lines. J. Immunol. Meth. 1993, 157: 233-240. 10.1016/0022-1759(93)90092-L.View ArticleGoogle Scholar
- Joubert R, Kuchler S, Zanetta JP, Bladier D, Avellana-Adalid V, Caron M, Doinel C, Vincendon G: Immunohistochemical localization of a β-galactoside-binding lectin in rat central nervous system. I. Light and electron-microscopical studies in developing cerebral cortex and corpus callosum. Dev. Neurosci. 1989, 11: 397-413.View ArticlePubMedGoogle Scholar
- Cornillot JD, Pontet M, Dupuy C, Chadli A, Caron M, Joubert-Caron R, Bourin P, Bladier D: Production and characterization of a monoclonal antibody able to discriminate galectin-1 from galectin-2 and galectin-3. Glycobiology. 1998, 8: 425-432. 10.1093/glycob/8.5.425.View ArticlePubMedGoogle Scholar
- Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature (London). 1970, 227: 680-685.View ArticleGoogle Scholar
- Berridge MV, Tan AS: Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT): subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Arch. Biochem. Biophys. 1993, 303: 474-482. 10.1006/abbi.1993.1311.View ArticlePubMedGoogle Scholar
- Scott DW, Livnat D, Pennell CA, Keng P: Lymphoma models for B cell activation and tolerance. III. Cell cycle dependence for negative signalling of WEHI-231 B lymphoma cells by anti-mu. J Exp Med. 1986, 164: 156-164.View ArticlePubMedGoogle Scholar
- Falini B, Pileri S, Pizzolo G, Durkop H, Flenghi L, Stirpe F, Martelli MF, Stein H: CD30 (Ki-1) molecule: a new cytokine receptor of the tumor necrosis factor receptor superfamily as a tool for diagnosis and immunotherapy. Blood. 1995, 85: 1-14.PubMedGoogle Scholar
- Tedder TF, Zhou LJ, Engel P: The CD19/CD21 signal transduction complex of B lymphocytes. Immunol Today. 1994, 15: 437-442. 10.1016/0167-5699(94)90274-7.View ArticlePubMedGoogle Scholar
- Sutherland R, Delia D, Schneider C, Newman R, Kemshead J, Greaves M: Ubiquitous cell-surface glycoprotein on tumor cells is proliferation-associated receptor for transferrin. Proc Natl Acad Sci USA. 1981, 78: 4515-4519.PubMed CentralView ArticlePubMedGoogle Scholar
- Serrano M, Hannon GJ, Beach D: A new regulatory motif in cell-cycle control causing specific inhibition of cyclin D/CDK4. Nature. 1993, 366: 704-707. 10.1038/366704a0.View ArticlePubMedGoogle Scholar
- Bender CM, Pao MM, Jones PA: Inhibition of DNA methylation by 5-aza-2'-deoxycytidine suppresses the growth of human tumor cell lines. Cancer Res. 1998, 58: 95-101.PubMedGoogle Scholar
- Chiariotti L, Berlingieri MT, Battaglia C, Benvenuto G, Martelli ML, Salvatore P, Chiappetta G, Bruni CB, Fusco A: Expression of galectin-1 in normal human thyroid gland and in differentiated and poorly differentiated thyroid tumors. Int J Cancer. 1995, 64: 171-175.View ArticlePubMedGoogle Scholar
- Ellerhorst J, Nguyen T, Cooper DN, Estrov Y, Lotan D, Lotan R: Induction of differentiation and apoptosis in the prostate cancer cell line LNCaP by sodium butyrate and galectin-1. Int J Oncol. 1999, 14: 225-232.PubMedGoogle Scholar
- Bouffard DY, Momparler LF, Momparler RL: Enhancement of the antileukemic activity of 5-aza-2'-deoxycytidine by cyclopentenyl cytosine in HL-60 leukemic cells. Anticancer Drugs. 1994, 5: 223-228.View ArticlePubMedGoogle Scholar
- Lutomski D, Bourin P, Bladier D, Caron M, Joubert-Caron R: Erythroid differentiation leads to an externalization of galectin1 from cytosol to cell surface of human erythroleukemia cell line. TF-1. Eur. J. Cell Biol. 1997, 20-Google Scholar
- Iglesias MM, Rabinovich GA, Ivanovic V, Sotomayor C, Wolfenstein-Todel C: Galectin-1 from ovine placenta – amino-acid sequence, physicochemical properties and implications in T-cell death. Eur J Biochem. 1998, 252: 400-407. 10.1046/j.1432-1327.1998.2520400.x.View ArticlePubMedGoogle Scholar
- Dore BT, Chomienne C, Momparler RL: Effect of 5-aza-2'-deoxycytidine and vitamin D3 analogs on growth and differentiation of human myeloid leukemic cells. Cancer Chemother Pharmacol. 1998, 41: 275-280. 10.1007/s002800050740.View ArticlePubMedGoogle Scholar
- Zinzar S, Silverman LR, Richardson EB, Bekesi G, Holland JF: Azacytidine plus verapamil induces the differentiation of a newly characterized biphenotypic human myeloid-B lymphoid leukemic cell line BW-90. Leuk Res. 1998, 22: 677-685. 10.1016/S0145-2126(98)00020-4.View ArticlePubMedGoogle Scholar
- Ahmed H, Sharma A, DiCioccio RA, Allen HJ: Lymphoblastoid cell adhesion mediated by a dimeric and polymeric endogenous beta-galactoside-binding lectin (galaptin). J. Mol. Recognit. 1992, 5: 1-8.View ArticlePubMedGoogle Scholar
- Baum LG, Seilhamer JJ, Pang M, Levine WB, Beynon D, Berliner JA: Synthesis of an endogeneous lectin, galectin-1, by human endothelial cells is up-regulated by endothelial cell activation. Glycoconj. J. 1995, 12: 63-68.View ArticlePubMedGoogle Scholar
- Levy G, Tarrab-Hazdai R, Teichberg VI: Prevention and therapy with electrolectin of experimental autoimmune myasthenia gravis in rabbits. Eur. J. Immunol. 1983, 13: 500-507.View ArticleGoogle Scholar
- Offner H, Celnik B, Bringman TS, Casentini-Borocz D, Nedwin GE, Vanderbark AA: Recombinant human β-galactoside binding lectin suppresses clinical and histological signs of experimental autoimmune encephalomyelitis. J. Neuroimmunol. 1990, 28: 177-184. 10.1016/0165-5728(90)90032-I.View ArticlePubMedGoogle Scholar
- Rabinovich GA, Daly G, Dreja H, Tailor H, Riera CM, Hirabayashi J, Chernajovsky Y: Recombinant galectin-1 and its genetic delivery suppress collagen-induced arthritis via T cell apoptosis. J. Exp. Med. 1999, 385-397. 10.1084/jem.190.3.385.Google Scholar
- Miyagawa J, Muguruma M, Aoto H, Suetake I, Nakamura M, Tajima S: Isolation of the novel cDNA of a gene of which expression is induced by a demethylating stimulus. Gene. 1999, 240: 289-295. 10.1016/S0378-1119(99)00450-3.View ArticlePubMedGoogle Scholar
- Biffen M, McMichael-Phillips D, Larson T, Venkitaraman A, Alexander D: The CD45 tyrosine phosphatase regulates specific pools of antigen receptor-associated p59fyn and CD4-associated p56lck tyrosine in human T-cells. Embo J. 1994, 13: 1920-1929.PubMed CentralPubMedGoogle Scholar
- Katagiri T, Ogimoto M, Hasegawa K, Mizuno K, Yakura H: Selective regulation of Lyn tyrosine kinase by CD45 in immature B cells. J. Biol. Chem. 1995, 270: 27987-27990. 10.1074/jbc.270.47.27987.View ArticlePubMedGoogle Scholar
- Weiss A, Schlessinger J: Switching signals on or off by receptor dimerization. Cell. 1998, 94: 277-280.View ArticlePubMedGoogle Scholar
- Vespa GNR, Lewis LA, Kozak KR, Moran M, Nguyen JT, Baum L, Carrie Micelli MC: Galectin-1 specifically modulates TCRsignals to enhance TCR apoptosis but inhibit IL-2 production and proliferation. J. Immunol. 1999, 162: 799-806.PubMedGoogle Scholar
- Perillo N, Pace KE, Seilhame JJ, Baum LG: Apoptosis of T-cells mediated by galectin-1. Nature. 1995, 378: 736-738. 10.1038/378736a0.View ArticlePubMedGoogle Scholar
- Gillenwater A, Xu XC, Estrov Y, Sacks PG, Lotan D, Lotan R: Modulation of galectin-1 content in human head and neck squamous carcinoma cells by sodium butyrate. Int J Cancer. 1998, 75: 217-224. 10.1002/(SICI)1097-0215(19980119)75:2<217::AID-IJC9>3.3.CO;2-N.View ArticlePubMedGoogle Scholar
- Kopper L, Sebestyen A: Syndecans and the lymphoid system. Leuk Lymphoma. 2000, 38: 271-281.View ArticlePubMedGoogle Scholar
- Poirier F, Pontet M, Labas V, le Caer JP, Sghiouar-Imam N, Raphael M, Caron M, Joubert-Caron R: Two-dimensional database of a Burkitt lymphoma cell line (DG 75) proteins: protein pattern changes following treatment with 5'-azycytidine. Electrophoresis. 2001, 22: 1867-1877. 10.1002/1522-2683(200105)22:9<1867::AID-ELPS1867>3.0.CO;2-7.View ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an Open Access article: verbatim copying and redistribution of this article are permitted in all media for any purpose, provided this notice is preserved along with the article's original URL.