Analysis of GLUT-1, GLUT-3, and angiogenic index in syndromic and non-syndromic keratocystic odontogenic tumors

Abstract The aim of this study was to evaluate the immunoexpression of glucose transporters 1 (GLUT-1) and 3 (GLUT-3) in keratocystic odontogenic tumors associated with Gorlin syndrome (SKOTs) and non-syndromic keratocystic odontogenic tumors (NSKOTs), and to establish correlations with the angiogenic index. Seventeen primary NSKOTs, seven recurrent NSKOTs, and 17 SKOTs were selected for the study. The percentage of immunopositive cells for GLUT-1 and GLUT-3 in the epithelial component of the tumors was assessed. The angiogenic index was determined by microvessel count. The results were analyzed statistically using the nonparametric Kruskal-Wallis test and Spearman’s correlation test. High epithelial immunoexpression of GLUT-1 was observed in most tumors (p = 0.360). There was a higher frequency of negative cases for GLUT-3 in all groups. The few GLUT-3-positive tumors exhibited low expression of this protein in epithelial cells. No significant difference in the angiogenic index was observed between groups (p = 0.778). GLUT-1 expression did not correlate significantly with the angiogenic index (p > 0.05). The results suggest that the more aggressive biological behavior of SKOTs when compared to NSKOTs may not be related to GLUT-1 or GLUT-3 expression. GLUT-1 may play an important role in glucose uptake by epithelial cells of KOTs and this process is unlikely related to the angiogenic index. GLUT-1 could be a potential target for future development of therapeutic strategies for KOTs.

Construction and identification of a lentiviral vector for RNA interference of human GLUT3 gene / 中南大学学报(医学版)

OBJECTIVE@#To construct an effective lentiviral vector for RNA interference (RNAi) with human glucose transporter 3 (GLUT3)gene.
@*METHODS@#Four pairs of shRNA sequences against different parts of GLUT3-mRNA were separately cloned into the RNAi plasmid vector pLV-shRNA by recombinant DNA technology to construct shRNA expression vectors pLV-shRNA-GLUT3-1, pLV-shRNA-GLUT3-2, pLV-shRNA-GLUT3-3, and pLV-shRNA-GLUT3-4. The vectors were transfected into HeLa cells to detect the effectiveness of GLUT3 gene silencing. One of effective vectors was selected and co-transfected into 293T cells with lentivirus packaging plasmids to obtain packaged lentivirus particles LV-GLUT3. After viral titer determination, U251 glioblastoma cells were infected with LV-GLUT3 at a multiplicity of infection (MOI) of 10. Finally, the expression of GLUT3 protein was detected by Western blot. 
@*RESULTS@#DNA sequencing demonstrated that the shRNA sequences were successfully inserted into the pLV-shRNA vectors. In HeLa cells, the expression of GLUT3-mRNA was significantly down-regulated by the recombinant vectors compared with negative control. The recombinant lentivirus LV-GLUT3 harvested from 293T cells had a titer of 1.5×10(9) TU/mL. After infection with LV-GLUT3, the expression of GLUT3 protein in U251 glioblastoma cells was down-regulated. 
@*CONCLUSION@#An effective lentiviral shRNA expression vector targeting the GLUT3 gene is successfully constructed and can be used for further study on the functions of GLUT3 gene.

Análise comparativa da imunoexpressão de GLUT-1, GLUT-3 e M-CSF em lesão periférica e central de células gigantes / Comparative analysis of GLUT-1, GLUT-3 and M-CSF immunoexpression in peripheral and central giant cell lesions

A lesão periférica de células gigantes (LPCG) e a lesão central de células gigantes (LCCG) são patologias histologicamente semelhantes que acometem a região de cabeça e pescoço. O estudo objetivou analisar a expressão imuno-histoquímica dos marcadores GLUT-1, GLUT-3 e M-CSF em uma série de casos de LPCG e LCCG, na tentativa de compreender os diferentes comportamentos biológicos destas entidades patológicas. A amostra foi constituída por 20 espécimes teciduais de LPCG, 20 de lesão central de células gigantes não agressivas (LCCGNA) e 20 de lesão central de células gigantes agressivas (LCCGA), oriundos do Serviço de Anatomia Patológica da Disciplina de Patologia Oral do Departamento de Odontologia da UFRN. Foi realizada a análise semiquantitativa e qualitativa da expressão imuno-histoquímica dos marcadores nas células gigantes e nas células mononucleares. Em relação ao GLUT-1, verificou-se uma diferença estatisticamente significativa na quantidade de células mononucleares imunomarcadas entre a LPCG e a LCCGNA e entre a LPCG e a LCCGA. Em relação à intensidade da marcação também foi verificado uma diferença estatisticamente significativa tanto para as células mononucleares quanto para as células gigantes entre LPCG e LCCGNA e entre LPCG e LCCGA, nas células gigantes também ocorreu uma diferença estatisticamente significativa entre a LCCGNA e a LCCGA. Em relação ao GLUT-3, foi encontrada uma diferença estatisticamente significativa entre LPCG e LCCGA e entre LCCGNA e LCCGA na quantidade de células mononucleares imunomarcadas. No que concerne à intensidade de marcação para a referida proteína foi verificado uma diferença estatisticamente significativa nas células gigantes entre LPCG e LCCGA. Para o M-CSF foi observada apenas uma diferença estatisticamente significativa na intensidade de marcação nas células mononucleares entre LPCG e LCCGNA e entre LPCG e LCCGA. Com base nestes resultados, pode-se concluir a participação do GLUT-1, GLUT-3 e do M-CSF na patogênese das lesões estudadas. A maior imunomarcação destas proteínas nas células mononucleares evidenciam que tais células desempenham uma maior atividade metabólica e osteoclastogênica, principalmente nas LCCGA. Constatou-se que as células mononucleares estavam mais relacionadas à patogênese das lesões estudadas do que propriamente as células gigantes (AU).

The peripheral giant cell lesion (PGCL) and the central giant cell lesion (CGCL) are lesions histologically similar affecting the head and neck region. The study aimed to analyze the immunohistochemical expression of markers GLUT-1, GLUT-3 and MCSF in a series of cases of PGCL and CGCL, in trying to understand the different biological behavior of these pathologies. The sample consisted of 20 tissue specimens of PGCL 20 central lesion of not aggressive giant cell (CLNAGC) and 20 central lesion of aggressive giant cell (CLAGC), coming from the Pathology Unit of Oral Pathology of the Department of Dentistry of UFRN. Was performed the semi-quantitative and qualitative analysis of immunohistochemical expression of the markers in giant cells and mononuclear cells. In relation to the GLUT-1, it was found a statistically significant difference (p <0.05) in the number of mononuclear cells immunomarked between the PGCL and the CLNAGC and between the PGCL and CLAGC. Regarding the intensity of staining was also observed a statistically significant difference both at the mononuclear cells as in giant cells between PL and CLNAGC and between PGCL and CLAGC, at the giant cells there was also a statistically significant difference between the CLNAGC and CLAGC. In relation to GLUT-3, was found a statistically significant difference between PGCL and CLAGC and between CLAGC and CLNAGC in amount of mononuclear cells immunomarked. Regarding the intensity of labeling for such protein was found a statistically significant difference at the giant cells between PL and CLAGC. To the M-CSF was observed only a statistically significant difference in the intensity of labeling at the mononuclear cells between PGCL and CLNAGC and between PGCL and CLAGC. Based on these results, we can conclude the participation of GLUT-1, GLUT-3 and M-CSF in the pathogenesis of the lesions studied. The bigger immunostaining of these proteins in mononuclear cells show that these cells perform a higher metabolic activity and osteoclastogenic, especially in CLAGC. It was found that the mononuclear cells were more related to the pathogenesis of the studied lesions than properly the giants cells (AU).

Expression of GLUT 3 in different brain regions of aged rats / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To examine the distribution of glucose transport 3 (GLUT 3) in different brain regions of aged rats and to investigate its role in ageing process of the nervous system.</p><p><b>METHODS</b>The GLUT 3 expression in different brain regions was examined with immunohistochemical method in rats aged 3, 18 and 30 months, respectively.</p><p><b>RESULTS</b>The number of GLUT 3-positive cells varied in the different brain regions in rats of all age groups (P<0.01); the CA1 region contained the greatest number of positive cells,and fewer in the motor cortex and cerebellum. The number of GLUT 3-positive cells was reduced in the brain of aged rats (P<0.01); and the neural cells in 4 different brain regions presented with large cell body and loose alignment.</p><p><b>CONCLUSION</b>The expression of GLUT 3 decreased in aged rats, which suggests that GLUT 3 may be involved in the ageing process of nervous system.</p>

Expression of glucose transporter-3 in the cerebral cortex of aging rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the changes in the expression of glucose transporter-3 (GLUT3) in the cerebral cortex of rats during aging and investigate the role of GLUT3 in the aging process of the nervous system.</p><p><b>METHODS</b>The cerebral tissues were collected from rats of 3, 18, 24, and 30 months old (10 in each age group), and the expression of GLUT3 in the cerebral cortex was detected by immunohistochemistry.</p><p><b>RESULTS</b>Under optical microscope, GLUT3-positive cells were found in every group. Within the age range of 3 to 8 months, GLUT3-positive cells increased significantly with age (P<0.01), but at 24-30 months of age, the number of GLUT3-positive cells reduced significant with age (P<0.01).</p><p><b>CONCLUSION</b>The expression changes of GLUT3 ir the cerebral cortex of rats during aging indicate that GLUT3 plays an important role in the maturation and aging of the nervous system.</p>

Dynamic expression of glucose transporters 1 and 3 in the brain of diabetic rats with cerebral ischemia reperfusion / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Blood glucose control improves the outcome of diabetic patients with stroke, but the target range of blood glucose control remains controversial. The functional recruitment of ischemia penumbra is extremely important to the recovery after stroke. The present study aimed to explore the expression of brain-type glucose transporters (GLUT1 and GLUT3) in cerebral ischemic penumbra at different blood glucose levels and different ischemic-reperfusion time in diabetic hypoxia-ischemia rats. The results might provide an experimental basis for clinical treatment of diabetic patients with stroke.</p><p><b>METHODS</b>The Wistar rats included in this study were randomly assigned to 4 groups (50 rats each): normal control group (NC), uncontrolled diabetic group (DM1), poorly-controlled diabetic group (DM2), and well-controlled diabetic group (DM3). Diabetic rats were induced by single intraperitoneal injection of streptozotocin, and the focal ischemic rat model of middle artery occlusion (MCAO) was made by insertion of fishing thread in 6 weeks after the establishment of the diabetic model. Each group was divided into 5 subgroups (10 rats each): four focal ischemic subgroups at different ischemic-reperfusion time (at 3,12, 24 and 72 hours after reperfusion, respectively) and one sham-operated subgroup. The mRNA and protein expression of GLUT1 and GLUT3 was assessed by RT-PCR and Western blotting, respectively.</p><p><b>RESULTS</b>There was significant difference in the mRNA expression of GLUT1 and GLUT3 between the four focal ischemic subgroups and the sham-operated subgroup at different reperfusion time in each group. The mRNA expression of GLUT1 and GLUT3 in the 4 ischemic groups began to increase at 3 hours, peaked at 24 hours after reperfusion and maintained at a higher level even at 72 hours compared with that of the sham-operated subgroup. The mRNA expression of GLUT1 increased more significantly than that of GLUT3. The mRNA expression of GLUT1 and GLUT3 was significantly different between the diabetic groups and normal control group. The mRNA expression of GLUT1 and GLUT3 was increased more significantly in the diabetic groups than that in the normal control group. There was a significant difference in the mRNA expression in the groups with different blood glucose levels. The mRNA expression tended to decrease with increased blood glucose levels. The expression trend of GLUT1 and GLUT3 protein was similar to that of GLUT1 and GLUT3 mRNA.</p><p><b>CONCLUSIONS</b>GLUT1 and GLUT3 expression was notably up-regulated in the penumbra region after cerebral ischemia in this study. But the up-regulated amplitude of GLUT1 and GLUT3 in the diabetic rats with cerebral ischemic injury became smaller than that of the normal controls. In the treatment of diabetic patients with cerebral embolism, blood glucose control should not be too strict, otherwise the up-regulation of GLUT1 and GLUT3 induced by cerebral ischemic injury might not be able to meet the needs of energy metabolism in cells.</p>

Expression of glucose transporter in non-small cell lung carcinoma and its clinical significance / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the expression of glucose transporter (Glut)1, Glut3, and hypoxia inducible factor (HIF)-1 alpha in human non-small cell lung carcinoma (NSCLC), and its clinical significance.</p><p><b>METHODS</b>Specimens of cancer tissues and paracancerous lung tissues from 34 cases of NSCLC and 17 specimens of benign lung lesions were collected. The expressions of Glut1, Glut3, and HIF-1 alpha were detected with immunohistochemical staining, RT-PCR, and Western blot.</p><p><b>RESULT</b>The relative mRNA expressions of Glut1 and HIF-1 alpha were 0.689 +/-0.245, 0.693 +/-0.248 in cancer tissues; and 0.338 +/-0.157, 0.351 +/-0.184 in paracancerous lung tissues (P <0.001); while those of Glut3 were 0.506 +/-0.246 in cancer tissues and 0.482 +/-0.238 in paracancerous tissues (P >0.05). The relative protein expressions of Glut1 and HIF-1 alpha were 0.582 +/-0.247, 0.525 +/-0.246 in cancer tissues and 0.288 +/-0.151, 0.261 +/-0.135 in paracancerous lung tissues (P<0.001), but the protein expressions of Glut3 were 0.551 +/-0.251 and 0.436 +/-0.224 respectively (P>0.05). Glut1 and HIF-1 alpha expressions were higher in poor differentiation group and in stage III group, than those in medium and well differentiation group and stage I and II group. Moreover, there was a significant correlation between the expression of Glut1 and HIF-1 alpha (r=0.854, P<0.01).</p><p><b>CONCLUSION</b>Glut1 and HIF-1 alpha are highly expressed in NSCLC, and their expressions are associated with tumor differentiation and clinical stage.</p>

Effect of progesterone on the expression of GLUT in the brain following hypoxic-ischemia in newborn rats / 中国应用生理学杂志

<p><b>AIM</b>To investigate the expression of GLUT1 and GLUT3 in the hippocampus after cerebral hypoxic-ischemia (HI) in newborn rats and the effect of progesterone (PROG) on them.</p><p><b>METHODS</b>Forty newborn SD rats were randomly divided into four groups: normal group, sham-operated group, hypoxic-ischemic group and progesterone group. Model of hypoxic-ischemia encephalopathy (HIE) was established in the 7-day-old newborn SD rats. Immunohistochemical method was applied to detect the expression of GLUT1 and GLUT3 in hippocampus.</p><p><b>RESULTS</b>GLUT1 and GLUT3 were slightly seen in normal and sham operation group, there was no obviously difference between the two groups (P > 0.05). The expression of GLUT1 and GLUT3 in hypoxic-ischemia group were all higher than that in sham operated group (P < 0.05). Not only the expression of GLUT in progesterone group were significantly higher than that in sham operated group (P < 0.01), but also than that in hypoxic-ischemia group (P < 0.05).</p><p><b>CONCLUSION</b>PROG could increase the tolerance of neuron to hypoxic-ischemia with maintaining the energy supply in the brain by up-regulating GLUT expression.</p>

Influence of blood glucose on the expression of glucose trans-porter proteins 1 and 3 in the brain of diabetic rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The delivery of glucose from the blood to the brain involves its passage across the endothelial cells of the blood-brain barrier (BBB), which is mediated by the facilitative glucose transporter protein 1 (GLUT(1)), and then across the neural cell membranes, which is mediated by GLUT(3). This study aimed to evaluate the dynamic influence of hyperglycemia on the expression of these GLUTs by measuring their expression in the brain at different blood glucose levels in a rat model of diabetes. This might help to determine the proper blood glucose threshold level in the treatment of diabetic apoplexy.</p><p><b>METHODS</b>Diabetes mellitus was induced with streptozotocin (STZ) in 30 rats. The rats were randomly divided into 3 groups: diabetic group without blood glucose control (group DM1), diabetic rats treated with low dose insulin (group DM2), and diabetic rats treated with high dose insulin (group DM3). The mRNA and protein levels of GLUT(1) and GLUT(3) were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry, respectively.</p><p><b>RESULTS</b>Compared with normal control rats, the GLUT(1) mRNA was reduced by 46.08%, 29.80%, 19.22% (P < 0.01) in DM1, DM2, and DM3 group, respectively; and the GLUT(3) mRNA was reduced by 75.00%, 46.75%, and 17.89% (P < 0.01) in DM1, DM2, and DM3 group, respectively. The abundance of GLUT(1) and GLUT(3) proteins had negative correlation with the blood glucose level (P < 0.01). The density of microvessels in the brain of diabetic rats did not change significantly compared with normal rats.</p><p><b>CONCLUSIONS</b>Chronic hyperglycemia downregulates GLUT(1) and GLUT(3) expression at both mRNA and protein levels in the rat brain, which is not due to the decrease of the density of microvessels. The downregulation of GLUT(1) and GLUT(3) expression might be the adaptive reaction of the body to prevent excessive glucose entering the cell that may lead to cell damage.</p>

Effect of different blood glucose levels on the expression of cerebral GLUT3 mRNA in neonatal rats with hypoxia-ischemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Concerns of the effect of glucose on perinatal hypoxic-ischemic brain damage are increasing. It was previously considered that the glucose transporter (GLUT) genes and their productions played an important role in the regulation of cerebral energy metabolism. The present study aimed to explore the effect of different blood glucose levels on the expression of cerebral GLUT3 mRNA in neonatal rats with hypoxia-ischemia (HI), and to evaluate the neuroprotective effect of glucose against HI insults.</p><p><b>METHODS</b>A total of 250 7-day-old neonatal SD rats were randomly divided into 10 groups (n=25 each): Normal control, Sham-operated, HI, Hypoglycemia, Hypoglycemia pre- and post-HI, Mild hyperglycemia pre- and post-HI, Severe hyperglycemia pre- and post-HI. Blood glucose levels of normal, hypoglycemia, mild hyperglycemia and severe hyperglycemia were defined as 5-7 mmol/L, 3-4 mmol/L, 10-15 mmol/L and 16-25 mmol/L, respectively. The expression of GLUT3 mRNA was detected with RT-PCT technique at 2, 24, 48 and 72 hrs and at 7 days after HI.</p><p><b>RESULTS</b>There was a correlation between increases in GLUT3 mRNA expression and postnatal age in the Normal control group. HI significantly enhanced the expression of GLUT3 mRNA from 2 hrs, peaking at 24 hrs after HI, and then significantly decreased at 72 hrs and 7 days after HI when compared with the Normal Control group (P < 0.01). GLUT3 mRNA expression in the Hypoglycemia pre-HI group was the lowest among all groups with HI at each time point after HI, and a statistically significant difference was found at 72 hrs after HI when compared with the HI group (P < 0.05). The expressional levels of GLUT3 mRNA in the Severe hyperglycemia pre-HI group were strikingly higher than those in any other groups with HI (P < 0.05 or 0.01). The GLUT3 mRNA expression patterns in the Mild and Severe hyperglycemia post-HI and the Hypoglycemia post-HI groups were similar to the Hypoglycemia pre-HI group.</p><p><b>CONCLUSIONS</b>GLUT3 mRNA expression and the synthesis of GLUT3 can be down-regulated by hypoglycemia pre-HI, coupled with aggravation of cerebral pathology, but up-regulated by higher hyperglycemia pre-HI, coupled with improvement of cerebral pathology. This suggested that adequate glucose supplement before HI can improve the cerebral function against HI insults in neonatal rats.</p>

CoCl2-induced enhancement of glucose transport activity in mediating hypoxic tolerance in cultured hippocampal neurons / 生理学报

The effect of CoCl(2) pretreatment on glucose transport activity of cultured newborn rat hippocampal neurons and its role in neuronal hypoxic tolerance were observed. The results showed that the 2-deoxy-D-[1-(3)H ]glucose uptake rate and the mRNA expressions of glucose transporters (GLUT1 and GLUT3) in the hippocampal neurons were significantly increased after a 24-hour pretreatment with CoCl(2). The cell injury induced by 6-hour or 8-hour hypoxic exposure was also greatly reduced by CoCl(2) pretreatment. The protective effect of CoCl(2) on the neurons was largely abolished by cytochalasin B, a specific inhibitor of glucose transporters. The results suggest that CoCl(2) can increase mRNA expressions of GLUT1 and GLUT3 and glucose transporter activity of the neurons, which may be an important mechanism for the increased tolerance of the neurons to hypoxia.