Micheline Soares Costa Oliveira 1, Thaís Rocha Cavalcante1, Victória Reggna Paulino Albuquerque 1, Victor Borges Fernandes1, Cristiane Duarte Alexandrino Tavares2
1 Química, Universidade Estadual Do Ceará (Ceará, Brasil), 2Química, Faculdade Cisne (Ceará, Brasil)
Different learning methods can be used to better assimilate program content. The biochemical mechanism of drugs in our body generally converts lipophilic chemical compounds into products more readily excreted. Its rate is an important determinant of the duration and intensity of the drug's pharmacological action. To show the effect of drugs on the body through play as a teaching- learning tool. For this understanding was applied a game for students of the 5th. Semester of Chemistry, in the discipline of Supervised Internship of the semesters of 2018.1 and 2019.2, totaling 24 participants. The game "Drugs Knowledge in Action"
consists of 110 cards. It consists of a deck with three types of cards, drug effects, drug image and drug chemical formula. Each participant (in all 4) received five cards, consisting of three sets of different drug titles. The winner was the one who formed the three sets first. Each participant resorted to the block of cards and retired until it had at least 9 cards in the hand, when exceeding that number, the other cards should be discarded. Biochemical action maps of these drugs were constructed throughout the game.
When questioned with the application of a quick QUIZ on the subject, 57% reponed that they were not aware of the chemical formula of the drugs in question. When it came to the effects of these drugs, 89% said they knew them, but some effects are common to two or more drugs. As for drug metabolism in the body only 35% of the participants knew this subject. The majority of the group 87% stated that they have clarified many doubts on the subject. .During the game participants exchanged many ideas and concepts about the metabolism of drugs. The applied game and other methodologies offer greater knowledge to the students and concretely strengthen their learning.
Keywords: Education and Biochemical, Games, Drugs
I – Glycobiology
I-01 - A Partial Kinetics Study for Chondroitinases AC
João Victor Gerheim da Silva 1, Matheus Miranda Fracetti1, Ana Luiza Carneiro da Silva1, Carlos Magno da Costa Maranduba 2, Yara Maria Corrêa da Silva Michelacci3, Jair Adriano Kopke de Aguiar1
1 Bioquímica, Universidade Federal de Juiz de Fora (MG, Brasil), 2Dep. de Biologia, Universidade Federal de Juiz de Fora (MG, Brasil),
3Bioquímica, Univ. Federal de São Paulo (MG, Brasil)
Chondroitinases (Chase) are enzymes widely used in the characterization of glycosaminoglycans (GAGs), a group of complex linear sulfated polysaccharides, found in a wide variety of tissues from many species. These GAGs have been implicated in cell-cell interaction binding with a variety of biologically important proteins. The aim of the present study was to investigate the mechanism related to GAG depolymerization catalyzed by Chase AC and obtain the catalytic parameters of this enzyme. Chase AC was isolated from a crude extract of Flavobacterium heparinum by Phenyl Sepharose FF hydrophobic interaction chromatography. The kinetic parameters were achieved, incubating 1UE of Chase AC with increasing substrate concentrations (either chondroitin 4- sulfate or 6-sulfate) and measuring the unsaturated products formed in a thermostabilized spectrophotometer at 232nm. Moreover, the products of enzymatic digestion were also analyzed by fluorophore-assisted carbohydrate electrophoresis (FACE). According to the substrate incubated, Chase AC appears to have a different Vmax and Km parameters. With a Vmax of 1.5x10 -3 when used C4S as substrate and 7,03x10 -3 when the substrate was C6S. The Km for each substrate was 8,02x10 +1 for C4S and 1,64 x10 +2 for C6S, meaning that the differences on its substrate structure, like the position of sulfate groups, have an impact on how the enzyme recognizes, interact and catalyze its substrates. We also found a different pattern on digestions when the products were analyzed using FACE. We were able to establish the kinetics parameters for each substrate such as its Km and Vmax, evidentiating the difference between the substrate and its catalysis.
Keywords: Chondroitinases, Glycosaminoglycans, Kinetics / Supported by: Nenhuma
I-02 - Clinical and Pre-Clinical Analysis of The Tumor Progression in Colorectal Cancer Under Diabetes
Ronan Santos1, Andréia Vasconcelos dos Santos1, César Bastos Junior3, Agatha Fonseca3, Miguel Fontes3, Wagner Dias1, Frederico Alisson1,2, Adriane Regina Todeschini 11Carlos Chagas Filho Institute of Biophysics, Federal University Rio de Janeiro (Rio de Janeiro, Brasil), 2Instituto de Microbiologia Professor Paulo de Góes, Federal University of Rio de Janeiro (RJ, Brasil), 3Patologia, Hospital Naval Marcílio Dias (Rio de Janeiro, Brasil)
Colorectal cancer (CRC) is the third leading cause of cancer-related death in Brasil and the most common malignancy in the western world. Epidemiological evidences show that individuals with diabetes mellitus (DM) have significantly higher risk of developing multiple types of cancers. Furthermore, DM correlates tightly with the incidence and mortality of CRC. Although there are many proposed mechanisms to explain these associations, more conclusive mechanisms are yet to be established. Previous studies from our group demonstrated that hyperglycemia exacerbates tumor malignancy through the hexosamine biosynthetic pathway (HBP) and aberrant glycosylation. In this study, we aim to evaluate if HBP influences CRC progression in diabetes-induced mice and diabetic patients. CDX2P-NLSCre Apc flox/+ (APC-CPC) mice, a mouse model of spontaneous CRC received STZ (50mg/kg) injection daily for 5 days to induce DM. The APC-CPC-DM mice were euthanized three months after DM induction and the tumor number, tumor load and colon length were evaluated. The density of tumor-infiltrating immune cells in tissue sections from diabetic patients was assessed by H&E-staining. Furthermore, the expression of the HBP limiting enzyme (GFAT) was analyzed by immunohistochemistry. The APC-CPC-DM presented increased tumor load compared to APC-CPC non-diabetic mice and this was accompanied by decreased in colon length. Tumor samples from patients with diabetes presents significant GFAT2 levels and had more significant tumor budding consistent with a more infiltrative profile. We conclude that diabetes can contributes for tumor malignancy and this can be associated with increased HBP influx.
Keywords: Colorectal câncer, diabetes, hexosamine biosynthetic pathway. Supported by: CNPq and CAPES
I-03 - Red Meat-Derived Glycan Ingestion Increases Signaling Pathways in Colorectal Cancer
Ana Luiza dos Santos Lopes 1, Wallace M. Araújo2, Miguel Clodomiro Lucena1, Jose Andres Morgado-Diaz2, Adriane Regina Todeschini1, Frederico Alisson Silva1
1 Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro (RJ, Brasil), 2Programa de Oncobiologia Celular e Molecular, Instituto Nacional do Câncer (RJ, Brasil)
Every living cell is covered by an extensive layer of glycoconjugates terminated by sialic acid, a monosaccharide that mediates many receptor/ligand interactions. While mammals synthesize the two isoforms of sialic acid Neu5Ac and Neu5Gc, humans do not synthesize Neu5Gc due to a mutation in the CMAH enzyme. Despite that, Neu5Gc has been found in many human tissues including colorectal carcinomas (CRC). Because ingestion of red meat is the only proposed source of Neu5Gc for humans and red meat consumption has been extensively associated with CRC risk, we wanted to investigate if the metabolic incorporation of Neu5Gc by human colon cells could affect signaling pathways relevant to growth or development of CRC. To test our hypothesis, HCT-116 human colorectal cell lines were fed with different concentrations of Neu5Gc and its metabolic incorporation was confirmed by HPLC and western blot. The activation of the Wnt and EGF signaling pathways as well as ligand binding to the cell surface were compared between Neu5Gc and Neu5Ac (control) fed cells. We found that human cells incorporated Neu5Gc in many cell surface glycoproteins, in a dose-dependent manner. When stimulated with Wnt3a, Neu5Gc-fed cells had increased response to both Wnt3a and EGF stimuli as observed by qRT-PCR and western blot. Further FACS analysis revealed that Neu5Gc
incorporation enhances the binding of Wnt3a to the cell surface suggesting that Neu5Gc incorporation can alter receptor/ligand interaction. We next aim to feed human-like Cmah-/- mice that spontaneously develop CRC with a Neu5Gc-rich diet to confirm if ingestion of this red meat derived glycan can influence CRC growth and progression in vivo. Our preliminary data suggests the dietary ingestion of Neu5Gc as a new mechanism involved in the progression of CRC associated to red meat consumption by humans. Keywords: Colorectal cancer, red meat-derived Neu5Gc, Wnt/EGF signaling pathways / Supported by: CAPES, FAPERJ and CNPq
I-04 - Production and Characterization of a Self-Crosslinking Chondroitin Sulfate with Great Potential for Hydrogel Formation
Ana Luiza C. da Silva 1, João Victor G. da Silva1, Arthur Girardi Carpanez2, Lenize Fernandes Maia3, Luiz Fernando C. de Oliveira3, Yara Maria C. Da Silva Michelacci4, Valquíria P. de Medeiros1, Jair Adriano K. de Aguiar1
1 Dep. de Bioquímica, Universidade Federal de Juiz de Fora (MG, Brasil), 2Dep. de Química, Universidade Federal de Juiz de Fora (MG, Brasil),
3Dep. de Química, Universidade Federal de Juiz de Fora (MG, Brasil), 4Dep. de Bioquímica, Universidade Federal de São Paulo (SP, Brasil) Chondroitin sulfate (CS) is a glycosaminoglycan with important biological functions, among them inflammation, cell proliferation, and tissue repair. Because of its excellent biocompatibility, it has a promising ability to simulate the native extracellular matrix for cell culture and regeneration of cartilaginous tissues. The objective of this study is the production and characterization of a self- crosslinking hydrogel from a chemical modification of CS for use as cells scaffold. The thiolated CS (CS-SH) was produced using different proportions of 1-Ethyl-3-(3-dimethyl aminopropyl) carbodiimide hydrochloride, N-hydroxysuccinimide, and cysteamine hydrochloride. Characterization of CS-SH was performed by agarose gel electrophoresis in propylene diamine acetate buffer (PDA), by chemical characterization (hexosamine, uronic acid, sulfhydryl groups) and spectroscopically (RAMAN, Infrared). The modal molecular weight of the samples was estimated by polyacrylamide gel electrophoresis. Metachromatic bands of CS and CS-SH samples were visualized in agarose gel electrophoresis. However, the band corresponding to the thiolated CS presented a differential migration than observed for the CS, due to the modification of the structure of the CS. The percentage of substitution ranged between 60% and 80%, by sulfhydryl groups and hexosamine estimation. Infrared spectroscopy of the modified product showed changes in the peak relative to the carboxylic acid near 1606 cm-1, the same was observed in RAMAN spectroscopy, which showed changes in the absorption near 1415 cm-1, were not identified in the CS spectrum pattern. Finally, the standard molecular weight of standard CS was estimated at 37 kDa whereas that of CS-SH(1) and CS-SH(2) was estimated at 113 kDa and 164 kDa. We have shown that the chemistry modification was efficient and the product has great potential for hydrogel formation.
Keywords: chondroitin sulfate, scaffold, glycosaminoglycan / Supported by: FAPEMIG
I-05 - Effect of sulphated polysaccharide in vascular dysfunction
Carlos Modesto Vera Palomino1, Renan Pelluzz Cavalheiro1, Luiz Fernando Braga patekoski1, Maria Cecília Zorél Menegletti1, Arthur Daniel Januzzi1, Paula Deboni1, Tábata Bergonci1, Gustavo Monteiro Vaina1, Helena Bonciani Nader1, Marcelo Andrade De Lima1
1 Bioquimica, Universidade Federal De São Paulo (SP, Brasil)
Sulphated polysaccharides (SPs) are a group of macromolecules found in different classes of organisms. The interest in these compounds is mainly due to its known pharmacological actions such as antioxidant, antitumor, anticoagulant, antithrombotic and antilipidemic activity. The role of SP in reactivity and glycocalyx recovery is known but its effect as a possible anti-atherogenic drug is unknown. Currently, there are several treatments for vascular dysfunction; however, they do have some adverse effects. Thus, the use of sulphated polysaccharides could be considered as an alternative for the treatment of vascular dysfunction such as atherosclerosis and hypertension. Evaluate the action of SP on the integrity and distribution of atherogenic plaques throughout the aorta of treated mice and their vasorelaxation activity in rat rings. Male mice (C57BL / 6 and ApoE -/-) received a 32% kcal atherogenic diet throughout 32 weeks combined with treatment with SPs and placebo. Then, they were euthanized, and their aorta analyzed by a combination of microscopy techniques. Confocal and Coherent Anti-Stokes Raman Scattering microscopy were used to characterize proteoglycans, lipids and collagen distribution. Standard optic microscopy (Hematoxylin/Eosin and Oil Red O staining) was used to study aorta integrity. Vasorelaxation was tested in rat aorta rings pre-treated with epinephrine and treated with
SPs in an organ bath. The experiments followed the ethics committee (CEUA N 8558170216). Differences in weight, plaque structure and distribution as well as aorta integrity were analyzed and compared between groups. SPs treatment reduced plaque formation and distribution throughout the aorta. Differences in aorta integrity were also found. SPs treatment reduced the atherogenic plaque formation hallmarks and induced vasorelaxant effects. SPs treatment attenuated the formation of atheroma plaque and it caused relaxation of the aorta rings.
Keywords: polysaccharide, glycocalyx, vascular dysfunction / Supported by: FAPESP, CNPq and CAPES
I-06 - The Interplay between O-GLCNAC and Phosporylation on Tyrosine Hydroxylase Activity and Cathecolamines Synthesis
Bruno da Costa Rodrigues 1, Anna Carolina Rêgo Costa1, Miguel Clodomiro dos Santos Lucena1, Isadora de Araújo Oliveira1, Danielle Beckman1, Sergio Teixeira Ferreira1, Fernando Garcia de Mello1, Adriane Regina Todeschini1, Ricardo Augusto de Melo Reis1, Wagner Barbosa Dias1
1 Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro (RJ, Brasil)
The Hexosamine biosynthetic pathway uses about 2% to 5% of all glucose that enters the cell, and has GFAT as the rate-limiting enzyme. This pathway has UDP-GlcNAc as final product, which is used as substrate for intracellular O-GlcNAc; a post-translational modification (PTM) resulting from the addition of GlcNAc on serine and/or threonine in proteins. This reaction is catalyzed by OGT, and the GlcNAc removal reaction is made by OGA. The balance of such enzymes will regulate the levels of O-GlcNAcylated proteins, which similarly to phosphorylation, are active in many cellular processes. Tyrosine hydroxylase (TH) is the rate-limiting step enzyme in catecholamine synthesis, hydroxylating L-tyrosine to obtain L-DOPA. Our aim is to investigate the role of O-GlcNAc on the tyrosine hydroxylase activity and cathecolamines synthesis. To do this, We used the pharmacological inhibitor of OGA, Thiamet-G; and nerve growth factor in PC12 cells, and performed western blot, immunoprecipitation, HPLC analysis and in vitro activity reactions. During NGF treatment, O-GlcNAc is modulated by OGA and GFAT downregulation. O-GlcNAcylation acts on the control of the phosphorylation levels on serine 40 in TH, where stimulation by 28% on the increase in phosphorylation at serine 40 decreases O-GlcNAc in 26%; while increase of intracellular O-GlcNAc in 19% reduces serine 40 phosphorylation by 16%. In addition, increasing the intracellular O-GlcNAcylation significantly reduces the levels of L-DOPA. Enzyme activity confirms that downregulation on TH's O-GlcNAc increases its activity by the increase on serine 40 phosphorylation. These data suggest a mechanism that integrates carbohydrate metabolism with the catecholamines pathway; where competition between O-GlcNAc and phosphate at serine 40 tyrosine hydroxylase site modulates its activity, controlling the synthesis of cathecolamine levels. In conclusion, O-GlcNAc is as relevant as phosphorylation on a dynamic tyrosine hydroxylase L-DOPA regulation, increasing the complexity of cathecolamines production control by a short-time PTM process.n
Keywords: O-GlcNAc, Tyrosine hydroxylase, Cathecolamines / Supported by: CNPq, CAPES, FAPERJ
I-07 - Biological Activities of the Sulfated Polysaccharides from the Green Algae Ulva lactuca L.
Rodrigo Costa Pacheco dos Santos 1, Natália Kelmer da Silva1, Rogéria Gabriela Campos de Andrade1, Tayná Rodrigues Coelho1, Helena Bonciani Nader2, Jair Adriano Kopke de Aguiar1, Valquiria Pereira de Medeiros1
1 Dep. de Bioquímica - ICB, Universidade Federal de Juiz de Fora (MG, Brasil), 2Dep. de Bioquímica - INFAR, Universidade Federal de São Paulo (SP, Brasil)
Seaweeds are promising sources of sulfated polysaccharides (SP) which have distinct monosaccharide composition, high
conformational complexity and high pharmacological potential To investigate SP anticoagulant and anti-inflammatory activities from green algae Ulva lactuca L. Ulva lactuca L. polysaccharides were obtained by enzymatic proteolysis and fractionation with different volumes of acetone (granted two fractions, F50Ul and F70Ul). F50Ul and F70Ul were analyzed by agarose gel electrophoresis, chemical dosages and FTIR/ATR. Monosaccharides composition were determined by Fluorophore-assisted Carbohydrate Electrophoresis. Anticoagulant activity in vitro was investigated by TTPa, TP, TT and anti-factor Xa and IIa. Fractions cytotoxicity were analyzed by MTT assay. Nitric oxide (NO) production was analyzed in rabbit aorta endothelial cells (RAEC) and RAW 264.7 macrophage LPS-induced by Griess. Metalloproteinases 2 and 9 (MMP-2,9) activity was investigated by zimogram. Pro-
inflammatory cytokines were measured by ELISA assay and the translocation of nuclear factor kappa B (NF-kB) using confocal microscopy. F50Ul and F70Ul presented metachromatic bands characteristic of polysaccharides rich in sulfate and this was confirmed by spectroscopic analyses. F50Ul and F70Ul present rhamnose as majority monosaccharide. F50Ul fraction prolonged the TTPa (0.1- 1.0µg/mL) and inhibited Xa and IIa factors activity while F70Ul showed no activity. F50ul and F70Ul wasn’t cytotoxicity by RAEC and RAW 264.7. RAEC treated with F50Ul (100µg/mL) increased NO production after 12 hours. F50Ul and F70Ul (12.5-25 µg/mL) reduced NO (17-21%, 58-70%, respectively) produced by macrophage LPS-induced. Also polysaccharides decreased the MMP-9 activity. F50Ul and F70Ul (100 µg/mL) reduced IL-6 (26%) and TNF-ɑ (18 and 26.6%). F70Ul (100 µg/mL) was able to reduce NF-kB translocation by the cytoplasm to nucleus in RAW 264.7 LPS- induced. Data suggests that F50Ul and F70Ul are molecules with promising biological activities. Therefore, F50Ul present an important anticoagulant activity in vitro while F70Ul has a substantial anti-inflammatory activity, probably related with the NF-kB pathway.
Keywords: Anticoagulant activity, Anti-inflammatory activity, Ulva lactuca L. / Supported by: UFJF, FAPEMIG, CAPES, CNPq.
I-08 - Isolation, Characterization and Anticoagulant/antithrombotic of Dermatan Sulfate from different origins.
João Victor Gerheim1, Iury Pitassi Zanon 1, Luiz Gustavo de Oliveira1, Arthur Girardi Carpanez1, Ana Luiza Carneiro da Silva1, Carlos Magno da Costa Maranduba1, Valquíria Pereira de Medeiros1, Yara Maria Corrêa da Silva Michelacci3, Jair Adriano Kopke de Aguiar1
1Bioquímica, 2Dep. de Biologia, Universidade Federal de Juiz de Fora (MG, Brasil), 3Bioquímica, Universidade Federal de São Paulo (SP, Brasil)
Dermatan sulfate (DS) is a glycosaminoglycan (GAG), formed by repeating units of disaccharide composed by iduronic acid or a glucuronic acid linked to an N-acetyl-galactosamine. DS structure is intrinsically heterogeneous and can interact with other structures on extracellular matrix components. DS could bind with heparin co-factor II (HCII) and exerts an
antithrombotic/thrombolytic activity. This study aimed to characterize the structure of DS from different origins and analyze its anticoagulant/antithrombotic activity. DS from different origins were extracted from porcine skin (PS) and bovine cornea (BC), proteolyzed with papain, precipitated with ethanol, purified by anion exchange chromatography, identified by agarose gel electrophoresis and characterized chemically (hexosamine, uronic acid, and sulfate contents), spectroscopically (InfraRed), and enzymatically through fluorophore-assisted carbohydrate electrophoresis. The DS-fractions where also tested to its
anticoagulant/antithrombotic capacity using aPTT, PT and fibrinogen essays. DS isolated from different origins and its fractions have a distinct polymeric structure, presenting a content of uronic acid and sulfate per hexosamine varying from each other. DS depolymerization with chondroitinases also shown a different pattern of disaccharides formed for each DS-fraction. DS from BC appears to have the lowest content of IdoA/Hexosamine, 0.6, when compared with PS fractions 1M and 2M, 1.1 and 1.6. It also appears to have the lowest ratio of sulfate/Hexosamine (0.8) when compared with 1.18 from PS-1M fraction and 1.11 from PS-2M fraction. The DS fractions PS-1M, PS-2M, and BC also presented an anticoagulant activity using aPTT assay increasing the normal clotting time (35s) at 1.7, 1.2 and 1.3 times, and fibrinogen test has increased the normal clotting time (9s) at 1.3, 2.0 and 1.2 times for PS 1M, PS 2M and BC, respectively. DS-fractions have not shown any difference on PT assay. We were able to isolate, characterize and assess the anticoagulant/antithrombotic activity of DS from different origins.
Keywords: Dermatan sulfate, Glycosaminoglycans, Extraction / Supported by: Nenhuma
I-09 - Evidences Of O-GLcNAc In Macroautophagy Of Glioblastoma
Amanda Vergueiro Leonel 1, Leonardo Travassos1, Adriane Regina Todeschini1, Kátia Carneiro2, Wagner B Dias1
1 Instituto de Biofísica Carlos Chagas Filho, 2Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro (RJ, Brasil)
Glioblastoma (GBM), the most aggressive glioma, is resistant to the chemotherapeutic Temozolamide (TMZ). Recent evidences suggest that autophagy is strongly related to GBM tumorigenesis, and both, tumorigenesis and autophagy have been associated with protein O-GlcNAcylation. O-GlcNAc is a post-translational modification catalyzed by O-GlcNAc transferase (OGT), while its removal is catalyzed by O-GlcNAcase (OGA). Pharmacological modulation of these two enzymes alters the autophagic flow in neuronal cells, but there is still no evidence of such modulation in GBM cells. Here, we hypothesized that O-GlcNAcylation participates in the regulation of autophagic flow and TMZ resistence in GBM cells. GBM cell lines, U87MG and GBM011, were incubated with Thiamet G (TMG, OGA inhibitor), Osmi-1 (OGT inhibitor), TMZ, rapamycin and chloroquine (positive and negative controls of autophagy). The protein levels of the autophagic markers, p62 and LC3, and the level of intracellular O-GlcNAcylated proteins were accessed by the Western Blotting assay. Viability and proliferation of GBM cells were analyzed by flow cytometry and trypan blue exclusion assay. OGA inhibition by TMG (1 µM) increased O-GlcNAcylation and autophagic flux, while OGA inhibition by Osmi-1 (50 μM) reduced LC3 expression and O-GlcNAcylation in GBM cells. In addition, TMG treatment promoted increased cell viability and proliferation, whilst Osmi-1 reduced cell viability more strongly than TMZ, also inducing cell cycle arrest in the G1 phase. Our results represent a step forward toward an understanding of O-GlcNAc role in autophagy, cellular viability and response to chemotherapy whit potential therapeutic implications. Keywords: O-GlcNAc, Autophagy, Glioblastoma / Supported by: CAPES, FAPERJ, CNPq and Cancer Fundation.
I-10 - Inhibition Of O-Glcnacylation Reduces Glioblastoma Cell Proliferation and Migration
Andreia de Vasconcelos dos Santos 1, Ana Rafaela Oliveira1, Hector Franco Loponte1, Bruna Mafra1, Luciana Romão1, Wagner Barbosa Dias1, Adriane Regina Todeschini1
1Instituto de Biofísica Carlos Chagas Filho, 2Instituto de Ciências Biológicas ICB universidade Federal do Rio de Janeiro (RJ, Brasil)
Glioblastoma (GBM) is the most common and aggressive malignant primary brain tumor. Epidemiological evidences show that individuals with diabetes mellitus (DM) have significantly higher risk of developing multiple types of cancers, include GBM. Our group have been proposed a mechanism to explain this association. We previously demonstrated that hyperglycemia is associated with increase flux through the Hexosamine Biosintetic Pathway (HBP) and consequently aberrant glycosylation, favoring tumor progression in colon cancer. Our objective is to investigate the aberrant glycosylation in astrocytes compared to GBMs cell lines and the contribution of different glucose concentration to malignancy Human GBMs and primary astrocyte were evaluated for their glycophenotypic profile. For the extracellular glycosylation, a panel of lectin binding assay was assessed. The intracellular glycosylation and HBP were identified by O-GlcNAc, OGA, OGT and GFAT1/GFAT2 levels, respectively. Moreover, we
pharmacologically inhibit the OGT and analyze the effect of this treatment in the migration and proliferation of GBMs Our data show that the GBMs cells line has higher O-GlcNAcylation when compared to normal astrocytes. In addition, the reduction of O-
GlcNAcylation levels by treatment with OGT inhibitor, promotes a significant decreased in the migration and proliferation of GBMs Our results show an important role of O-GlcNAcylation to GBMs migration and proliferation indicating a promising anti-tumoral approach. Keywords: GBM, High glucose, O-GlcNAcylation / Supported by: CAPES, CNPq