The pentose phosphate pathway (PPP; also called the phosphogluconate pathway and the hexose monophosphate shunt) is a process that breaks down glucose-6-phosphate into NADPH and pentoses (5-carbon sugars) for use in downstream biological processes. Micronodular tumor burden was assessed by measuring the cumulative micronodular tumor area at 4 coronal levels. Next lesson. 14 of Principles of Biochemistry, “Glycolysis, Gluconeogenesis, & Pentose Phosphate Pathway.” PENTOSE PHOSPHATE PATHWAY This pathway produces ribose from glucose, and it also generates 2 NADPH. Likewise, in glioblastoma tissue sections, PPP enzymes are strongly expressed in highly proliferative regions but are downregulated in severely hypoxic pseudopalisades, while glycolysis enzymes exhibit an inverse pattern.7. The pentose phosphate pathway is an alternative to glycolysis. GLYCOLYSIS, GLUCONEOGENESIS, AND THE PENTOSE PHOSPHATE PATHWAY. The pentose phosphate pathway is a metabolic pathway that takes place parallel to glycolysis. KC
While the importance of the PPP for rapid cell proliferation is rather obvious since the PPP generates building bricks and reductive power to fuel anabolic processes, the association between glycolysis and cell migration is less straightforward to explain. Tumor cell proliferation (Ki-67) was increased in shALDOC_1 tumors. After 3 days of hypoxic incubation, proliferation of all cell types was significantly decreased compared with normoxic controls (Fig. Animals were killed using CO2, brains were fixed in formalin and embedded in paraffin, and serial sections were stained with H&E. We investigated whether the 2 parallel glucose metabolic pathways are intrinsically linked with cell function and whether these pathways are mechanistically involved in regulating functional programs. Key Terms. In addition, diffusely infiltrative tumor growth was quantified by measuring the net area occupied by tumor cell nuclei in 20 representative regions using a previously developed method.9,13,14 Neither the analysis of micronodular tumor growth nor quantification of diffusely invasive growth revealed a significant difference between shALDOC_1 and shControl tumors (Fig. Digital images of all 20 regions were acquired from H&E-stained sections, transformed into black and white, and the percentage of black pixels was quantified. , Bottoni P, Pontoglio A, Mastrototaro L, Giardina B. Semenza
We next analyzed whether the hypoxia-induced shift from proliferation to migration is limited to GS cells or if it is a common phenomenon. glycolysis: The cellular degradation of the simple sugar glucose to yield pyruvic acid and ATP as an energy source. Since associations between metabolic and functional state are obviously independent of changes in oxygen tension, we asked whether different glucose pathways are mechanistically involved in the dichotomous regulation of cell motility and growth. A
Immunostaining of paraffin sections for Ki-67 was performed as described previously using a mouse monoclonal antibody (Dako,) and the Histofine detection system (Medac).13 The percentage of Ki-67 immunoreactive nuclei was determined in 3 high-power fields in the most actively proliferating tumor area. Tumor tissue from the cancer patients was included in the study. Abbreviations: HK2, hexokinase 2; GPI, glucose-6-phosphate isomerase; PFKP, 6-phosphofructokinase platelet type; ALDOC, aldolase C; TPI1, triosephosphate isomerase 1; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; PGK1, phosphoglycerate kinase 1; PGAM1, phosphoglycerate mutase 1; ENO1, enolase 1; PKM2, pyruvate kinase M2; LDHA, lactate dehydrogenase A chain; G6PD, glucose-6-phosphate dehydrogenase; PGLS, 6-phosphogluconolactonase; PGD, 6-phosphogluconate dehydrogenase; TKT, transketolase; TALDO1, transaldolase 1. U
or. The endpoint of the G55 xenograft model was the occurrence of symptoms such as weight loss ≥10% or neurological symptoms. Create a free account to download. There are two distinct phases in the pathway: the oxidative phase and the non-oxidative phase. NADPH is required for and consumed during fatty acid synthesis and the scavenging of … https://youtu.be/30bRJwBqykw~*~Hey! 1A and B). Two novel cell culture models of buccal mucosal oral cancer from patients with no risk-habits of tobacco smoking or chewing. After 6 hours (G55) or 28 hours (GS-11) of incubation, nonmigrated cells from the upper side of the transwell membrane, as well as migrated cells from the underside, were scraped off and collected separately. . Downregulation of G6PD was successful with 2 shRNAs in GS-11 and 3 in G55 cells (Fig. Tumor xenografts were generated as described previously.3 Briefly, dissociated G55 cells (4 × 104) or GS-11 cells (1.5 × 105) in 4 µL medium were injected stereotactically into the striatum of 6–8 week old anesthetized NMRI/Foxn1nu mice or SHrNTM Hairless NOD.SCID mice, respectively (Harlan Laboratories). For inhibitor experiments, 6-aminonicotinamide (6-AN) or 2-deoxyglucose (2-DG) were added to the upper and lower wells. To substantiate these observations, expression of ALDOC and G6PD were knocked down. . Agnihotri
, Walenta S, Ksiazkiewicz M, Kunz-Schughart LA, Mueller-Klieser W. Fairbank
Expression of almost all glycolysis enzyme transcripts was significantly increased in migratory cells compared with nonmigrated cells, whereas PPP enzyme expression was regulated inversely (Fig. Kathagen
Meaning of Pentose Phosphate Pathway. The finding that hypoxia increased the migration of epithelial cancer cell types supports the notion that hypoxia acts as a major driver of cancer progression by provoking acquisition of invasive and metastatic properties.16 The observation that migration of non-neoplastic cells prevalent in adult brain (including astrocytes, MSCs, fibroblasts, and mononuclear cells) was also accelerated by hypoxia indicates that enhanced movement of stromal cells governed by hypoxic tumor gradients may foster mutual stimulatory interactions between tumor and stromal cells in glioblastomas. So it is a shunt of glycolysis, Difference between batch, fed-batch and continuous culture technique, Naveed ali biotecnologest Finally, we explored whether glycolysis and the PPP are directly and causatively involved in the dichotomous regulation of migration versus proliferation. We addressed this question by using enzyme inhibitors and enzyme knockdown of G6PD and ALDOC. For full access to this pdf, sign in to an existing account, or purchase an annual subscription. G6PD is the first and key rate-controlling enzyme of the PPP, and G6PD and ALDOC are the strongest inversely hypoxia-regulated enzymes of the PPP and glycolysis in GS lines7 (Fig. Glioblastoma cell lines grown under adherent conditions (U87, G55), cell lines derived from other tumor entities (HuH7, MDA-MB-231), and non-transformed cells such as normal human astrocytes, fibroblasts, umbilical vein endothelial cells, glioblastoma-associated mesenchymal stem cells (MSCs), and peripheral blood mononuclear cells were included. Western blot analysis was performed as detailed in the Supplementary material and as described previously.7 Western blot scans of x-ray films were quantified by densitometry using the ImageJ program. For permissions, please e-mail: journals.permissions@oup.com. Values in (C)and (D) are means ± SD of sextuplicate determinations. 6B). Practice: Glucose-6-phosphate dehydrogenase and the most important function of the pentose phosphate pathway. . Whereas mice engrafted with G6PD knockdown cells survived longer, animals with ALDOC knockdown tumors died sooner than controls. , Laabs Y, Gunther HSet al. Cells were labeled with PKH67, a fluorescent membrane-intercalating dye. While HMP shunt is the breakdown of glucose to yield different metabolic intermediates. Two Phases: [1] Oxidative Phase & [2] Non-oxidative Phase Glucose 6-Phosphate + 2 NADP+ + H 2 O Ribose 5-Phosphate + 2 NADPH + CO 2 + 2H+ All experiments were performed at 21% O2. In most organisms, glycolysis occurs in the cytosol. While the pentose phosphate pathway does involve oxidation of glucose, its primary role is anabolic rather than catabolic. ME
Phases. All rights reserved. , Martens T, Matschke Jet al. In the GS-11 model, knockdown of G6PD had no significant effect on micronodular or diffusely invasive growth; however, proliferation rates were decreased by 3.6% compared with controls (P < .05), (Fig. Immunohistochemically, reduced expression of ALDOC was detected in shALDOC_1 tumors compared with shControls. . Heptose (sedoheptulose) Further metabolism. In glioblastoma stem-like cells, hypoxia induces downregulation of pentose phosphate pathway (PPP) enzymes and a flux shift towards glycolysis. DJ
Sextuplicates of 1.5 × 104 cells in 50 mL assay medium were seeded into the upper wells. FIGURE 14-27 Role of NADPH in regulating the partitioning of glucose 6-phosphate between glycolysis and the pentose phosphate pathway. The ED pathway involves an initial phosphorylation as in glycolysis but then is followed by an oxidative step of the compound to an acid (phosphogluconic acid). It furthers the University's objective of excellence in research, scholarship, and education by publishing worldwide, This PDF is available to Subscribers Only. It also oxidize glucose to CO2 and water. 4A). [14C]Glucose counts retained by the tissue, glycolysis, and pyruvate formation were also measured. In the ED pathway, glucose is converted to pyruvic acid in fewer steps than it is in the pathway of glycolysis. Giese
Enzyme expression, migration, and proliferation under hypoxia were studied in multiple cell types. Expression of glycolysis and pentose phosphate pathway (PPP) enzymes in G55 cell subpopulations. In conclusion, our findings indicate that inhibition of G6PD and the PPP could be a useful strategy for targeting highly proliferative tumor cells in glioblastoma. READ PAPER. , Espedal H, Keunen Oet al. 2B, Supplementary material, Fig. Pentose phosphate pathway Overview. Time points for FACS-separation of fast dividing (PKHlow) and slowly dividing (PKHhigh) subpopulations were optimized for each line to maximize differences in fluorescence intensity as well as population size for further analyses and were 2 days for G55 cells and 14 days for GS-11 cells. Optimal time points for separation of fast dividing (PKHlow) and slowly dividing (PKHhigh) populations were determined in pilot experiments to maximize differences in fluorescence intensity as well as in population size for each cell type. Proliferation and migration were analyzed in the presence or absence of 6-AN (A) or 2-DG (B). M. Vinayagam. 2-DG is a glucose analog that inhibits HK, the rate-limiting enzymatic step of glycolysis that is also essential for fueling the PPP with glucose-6-phosphate (Fig. S
Antiangiogenic treatment can inhibit glioblastoma growth in orthotopic xenograft models and reduce tumor cell proliferation; however, treated tumors typically display a striking compensatory increase in invasion.2,3 Inhibition of angiogenesis impairs intratumoral oxygen supply, and hypoxia is a potent driver of glioblastoma cell migration and invasion.4 Hypoxia can further enhance stem cell characteristics of glioblastoma cells, and cyclic fluctuations in oxygen tension, which occur during tumor progression, select for stem-like glioblastoma cells which possess outstanding metabolic adaptability.5,6 While investigating mechanisms of adaptation of glioblastoma stem-like (GS) cells to bidirectional fluctuations in oxygen tension, we recently discovered a reciprocal metabolic switch between the pentose phosphate pathway (PPP) and glycolysis.7 The PPP, which produces ribose-5-phosphate and NADPH for DNA/RNA and fatty acid synthesis, is an alternative anabolic pathway to the preparatory phase of glycolysis.8 Enzymes of the PPP are highly expressed under normoxic conditions, whereas acute hypoxia causes downregulation of PPP enzymes concomitant with upregulation of glycolysis enzymes. Exemplary analysis of diffuse invasion is shown for L2 (anterior commissure level). Pentose (ribose, xylulose, ribulose) Hexose (glucose, fructose) and. The pentose phosphate pathway is an alternative path for the oxidation of glucose. , Stracke ML, Liotta LA, Schiffmann E. Shiraishi
P
Cell cultures were established from human tissue or blood samples with informed consent of the patients and approval by the local ethics committees in Hamburg, Munich, and Heidelberg. Wang
4. End product of pentose phosphate pathway enter into glycolysis. Immunohistochemical analyses showed that the expression of glycolysis enzymes is strongly increased in pseudopalisading cells, whereas PPP enzyme expression is reduced.7 Pseudopalisading cells most likely represent a wave of tumor cells that actively migrate away from a severely hypoxic necrotic area arising after a microvascular insult.17 In contrast, expression of PPP enzymes is elevated in highly proliferative tumor regions where expression of glycolysis enzymes is comparatively low. (B) Migrated (M) and nonmigrated (N) cells were separated using transwell assays. , Schmidt NO, Eckerich Cet al. Ideally, both pathways should be blocked simultaneously to target both the highly proliferative as well as the invasive tumor component; this might be achieved by inhibiting HK2, for example by using inorganic phosphate or azole antifungal agents as reported recently by others.26,27 HK2 catalyzes an irreversible reaction essential for both glycolysis as well as the PPP. Glucose 6-phosphate for the production of NADPH (by the reduction of coenzyme NADP) and a Pentose sugar.. Effect of hypoxia on enzyme expression and cell function. Conversely, glycolysis enzymes were elevated in migrating cells, whereas PPP enzymes were diminished. Asterisks indicate significant maximal upregulation or downregulation of transcripts, which typically occurred at 48 hours (P < .05). Search for other works by this author on: Cost of migration: invasion of malignant gliomas and implications for treatment, Anti-VEGF treatment reduces blood supply and increases tumor cell invasion in glioblastoma, Inhibition of glioma angiogenesis and growth in vivo by systemic treatment with a monoclonal antibody against vascular endothelial growth factor receptor-2, Bevacizumab treatment induces metabolic adaptation toward anaerobic metabolism in glioblastomas, CD133 is a marker of bioenergetic stress in human glioma, The hypoxic microenvironment maintains glioblastoma stem cells and promotes reprogramming towards a cancer stem cell phenotype, Hypoxia and oxygenation induce a metabolic switch between pentose phosphate pathway and glycolysis in glioma stem-like cells, Understanding the Warburg effect: the metabolic requirements of cell proliferation, A distinct subset of glioma cell lines with stem cell-like properties reflects the transcriptional phenotype of glioblastomas and overexpresses CXCR4 as therapeutic target, Ku70/80 gene expression and DNA-dependent protein kinase (DNA-PK) activity do not correlate with double-strand break (dsb) repair capacity and cellular radiosensitivity in normal human fibroblasts, Isolation and culture of human neuromicrovascular endothelial cells for the study of angiogenesis in vitro, A novel one-armed anti-c-Met antibody inhibits glioblastoma growth in vivo, Inhibition of glioblastoma growth in a highly invasive nude mouse model can be achieved by targeting epidermal growth factor receptor but not vascular endothelial growth factor receptor-2, Inhibition of intracerebral glioblastoma growth by targeting the insulin-like growth factor 1 receptor involves different context-dependent mechanisms, Glycolytic enzyme inhibitors in cancer treatment, The hypoxic tumor microenvironment: A driving force for breast cancer progression, Pseudopalisades in glioblastoma are hypoxic, express extracellular matrix proteases, and are formed by an actively migrating cell population, A G{alpha}i-GIV molecular complex binds epidermal growth factor receptor and determines whether cells migrate or proliferate, microRNA-451: A conditional switch controlling glioma cell proliferation and migration, The “go or grow” potential of gliomas is linked to the neuropeptide processing enzyme carboxypeptidase E and mediated by metabolic stress, EphB2 receptor controls proliferation/migration dichotomy of glioblastoma by interacting with focal adhesion kinase, Proteomic characterization of harvested pseudopodia with differential gel electrophoresis and specific antibodies, Glycolysis as primary energy source in tumor cell chemotaxis, Glycolysis is the primary bioenergetic pathway for cell motility and cytoskeletal remodeling in human prostate and breast cancer cells, Lactate enhances motility of tumor cells and inhibits monocyte migration and cytokine release, The complex biology of autocrine motility factor/phosphoglucose isomerase (AMF/PGI) and its receptor, the gp78/AMFR E3 ubiquitin ligase, Identification of the azole class of antifungals as potent inhibitors of hexokinase II mediated tumour metaboism in glioblastoma, Hexokinase 2 is required for tumor initiation and maintenance and its systemic deletion is therapeutic in mouse models of cancer. Real-time PCR analyses were performed as detailed in the Supplementary material and as described previously,7 using validated TaqMan Gene Expression Assays and a 7500 Fast Real-Time PCR System (Applied Biosystems). , Verdone JE, Huang Jet al. qPCR and immunoblot analysis revealed downregulation of glycolysis enzymes in fast dividing (F) versus slowly dividing (S) cells, paralleled by upregulation of PPP enzymes. Immunohistochemistry for Ki-67 revealed that the tumor cell proliferation was 16.4% higher in shALDOC_1 tumors compared with shControls (P < .05). , Plottner N, Johansen J, Overgaard J, Dikomey E. Lamszus
G55 cells exhibit a very high proliferation rate in vitro and in vivo and form rapidly expanding, noninvasive nodular tumors in the mouse brain. The pentose phosphate pathway produces NADPH for fatty acid synthesis. None declared. J
(A) Fast and slowly dividing cell subpopulations were isolated by fluorescence-activated cell sorting (FACS) from PKH67-labelled cells (slowly: 8.9%, fast: 9.4% of all cells). These findings support the results of the enzyme inhibitor experiments and provide evidence that glycolysis and the PPP are mechanistically relevant for the dynamic regulation of migration versus proliferation. Indeed, the reactions that constitute glycolysis and its parallel pathway, the pentose phosphate pathway, occur metal-catalyzed under the oxygen-free conditions of the Archean oceans, also in the absence of enzymes. The pathway analysis will include glycolysis and pentosephosphate flux comparison between responders and non-responders during the recovery induced by mechanical unloading. To assess whether changes in gene expression corresponded to protein levels, immunoblot analyses were performed after 48 hours, and the results consistently confirmed that hypoxia induced downregulation of PPP enzyme expression, concomitant with upregulation of glycolysis enzymes (Fig. in glycolysis, CO2 is released from the mitochondria whereas in pentose phosphate pathway, CO2 is released in the cytosol Q. how does NADPH protect red blood cells from hemolysis? , Bronisz A, Nowicki MO, Chiocca EA, Lawler S. Horing
Krebs (citric acid) cycle and oxidative phosphorylation. De-coding group 3 medulloblastoma biology with non-coding RNA, Role of surgery for glioblastoma: response to letters from Dr. Gerritsen and his colleagues and Dr. Vargas Lopez, Therapeutic targeting of transcriptional elongation in diffuse intrinsic pontine glioma, Stemming the growth of pediatric gliomas through histone modification, In search of predictive and response markers in antiangiogenic therapy of glioblastoma, About the Japan Society for Neuro-Oncology, About the European Association of Neuro-Oncology, Receive exclusive offers and updates from Oxford Academic, Radiological assessment schedule for high-grade glioma patients during the surveillance period using parametric modeling, TSPO deficiency induces mitochondrial dysfunction, leading to hypoxia, angiogenesis, and a growth-promoting metabolic shift toward glycolysis in glioblastoma, The secreted glycolytic enzyme GPI/AMF stimulates glioblastoma cell migration and invasion in an autocrine fashion but can have anti-proliferative effects, Biological and therapeutic implications of multisector sequencing in newly diagnosed glioblastoma. It generates NADPH and pentoses as well as ribose 5-phosphate, a precursor for the synthesis of nucleotides. Patra
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, Cantley LC, Thompson CB. , Vartanian A, Burrell Ket al. G55 cells typically form highly proliferative, but largely noninvasive tumors, which are usually fatal within 3 weeks.3 In contrast, GS-11 tumors are highly invasive but grow slowly, so that development of tumor-related symptoms usually takes at least 7 months.7 Mice with orthotopic G55-shALDOC_1 xenografts had a significantly shorter survival (median, 17 d) than animals with shControl xenografts (median, 19 d), (P < .05), (Fig. . , St-Pierre P, Nabi IR. GS-11 and GS-12 cell lines were cultured as neurospheres, as described previously.9 Glioma cell lines G55 and U87, HuH7 (hepatocarcinoma), and MDA-MB231 (breast cancer) cells were cultured in Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum (FBS; Life Technologies). NADPH is generated in the oxidative phase, while pentose sugars are generated through the non-oxidative phase. Survival analyses were performed using the MedCalc program (Kaplan-Meier analysis, log-rank test). Glycolysis (also called Embden Meyerhoff pathway) and HMP shunt (also called pentose phosphate pathway, phosphogluconate pathway or direct pathway) both are glucose breakdown pathways. Cells from 6 transwell assays were pooled for RNA and protein extraction. Quantification of invasive growth in the GS-11 model was performed as described previously9,13,14 and as detailed in the Supplementary material. 2A, Supplementary material, Fig. E
S3B). 4B). Brat
5B). Copyright © 2021 Society for Neuro-Oncology. For example, glucose 6-phosphate isomerase (GPI) is identical to the secreted cytokine autocrine motility factor (AMF) which binds to a cell surface receptor, AMFR, and stimulates migration in an autocrine and paracrine fashion.26 The multifaceted “moonlighting” functions of glycolysis enzymes are only partly elucidated, and further work is necessary to better characterize their involvement in cancer cell function. Furthermore, migration of all cell lines was also inhibited by 2-DG, and the effects became significant at 0.5 or 1.0 µM (Fig. However, some mutations that disrupt the plastidial oxidative pentose phosphate pathway (OPPP) cause developmental defects, as well as embryo arrest at the globular stage of development. To quantify the extent of this type of invasive growth, the cumulative micronodular area on 4 different defined coronal sections was measured. 6-AN reduced glioblastoma cell proliferation while stimulating migration, whereas 2-DG reduced both proliferation and migration, indicating that interference at the enzymatic branching point that supports both pathways affects both associated functional states. Values are means ± SD of sextuplicate determinations. . Knockdown of G6PD resulted in decreased cell proliferation. Cellular respiration introduction. Goetze
. 5C). (A) Quantification of glycolysis and pentose phosphate pathway (PPP) enzyme transcripts by qPCR in different cell types exposed to hypoxia (H), (1% O2). A key intermediate in the breakdown of fructose obtained from the diet is fructose-1,6-bisphosphate fructose-6-phosphate fructose-2,6-bisphosphate fructose-1-phosphate UDP-fructose To determine whether glycolysis and the PPP are causatively involved in the dichotomous regulation of migration versus proliferation, we used enzyme inhibitors and shRNA. . , Schulte A, Balcke Get al. The pentose phosphate pathway is a metabolic pathway parallel to glycolysis. . Beckner
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. . Conversely, cell migration was found to be invariably enhanced by incubation under hypoxia for 5 hours in modified Boyden chamber assays (Fig. The difference is that glycolysis is the breakdown of glucose up to pyruvate or lactate. Hypoxia further causes increased GS cell migration but with reduced proliferation, whereas oxygenation has opposite effects linking the metabolic switch to the go or grow potential of the cells. There are two distinct phases in the pathway. We further observed a uniform regulation of go versus grow cellular programs. ALDOC knockdown resulted in significantly reduced migration of all GS-11 and G55 sublines (Fig. Glucose-6-phosphate dehydrogenase (G6PD) and Aldolase C (ALDOC), the most strongly inversely regulated PPP and glycolysis enzymes, were knocked down by short hairpin RNA. We previously discovered a metabolic switch in glioblastoma stem-like cells from the PPP to glycolysis and vice versa in response to hypoxia and oxygenation.7 Our current findings extend these observations by showing that the switch is not limited to GS cells but is a common phenomenon that also occurs in other types of cancer cells as well as in non-neoplastic cells that do not exhibit the Warburg phenotype. Annegret Kathagen-Buhmann, Alexander Schulte, Jonathan Weller, Mareike Holz, Christel Herold-Mende, Rainer Glass, Katrin Lamszus, Glycolysis and the pentose phosphate pathway are differentially associated with the dichotomous regulation of glioblastoma cell migration versus proliferation, Neuro-Oncology, Volume 18, Issue 9, September 2016, Pages 1219–1229, https://doi.org/10.1093/neuonc/now024. qPCR and immunoblot analysis revealed that PKHlow cells displayed decreased expression of almost all glycolysis enzymes and of LDHA compared with PKHhigh cells but increased expression of PPP enzymes, including its first and key regulatory enzyme G6PD (Fig. Sort by: Top Voted. Heddleston
6B). Functional effects of 6-AN and 2-DG. . , Gunther HS, Phillips HSet al. ME
On cross sections, perivascular invasion typically appeared as micronodular or finger-like structures. Rapidly and slowly dividing or migrating glioblastoma cells were separated, and enzyme profiles were compared. glucose-6-phosphate … G6PD and ALDOC emerged as the most strongly inversely hypoxia-regulated enzymes of both pathways. In the present study, we investigated whether the hypoxia-induced reciprocal switch between PPP and glycolysis enzyme expression is restricted to stem-like glioblastoma cells or if it also occurs in other cell types. . Incubation with 6-AN, an antimetabolite and competitive inhibitor of G6PD, reduced the proliferation of GS-11, GS-12, and of G55 cells in a dose-dependent fashion with >50% inhibition at 10–20 µM in all lines (Fig. Godlewski
Migration of GS-11 cells was decreased maximally by 70.5% (shALDOC_3), and G55 cell migration was reduced by up to 48.3% (shALDOC_1) compared with nonsilencing controls (shControl), (P < .05). , Chen X, An J, Day BW, Pollack IF. . Kasten
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The effect of G6PD knockdown on the in vivo growth of G55 tumors was contrary to that of ALDOC knockdown; ie, the survival of animals engrafted with G55- shG6PD_2 cells was prolonged (median, 20.5 d) compared with shControls (median, 18.5 d), (P < .05). (D) G6PD expression was reduced in GS-11-shG6PD_2 tumors. , Castellano-Sanchez AA, Hunter SBet al. Gene expression values are means ± SD of triplicate determinations, asterisks indicate significant downregulation compared with shControls (P < .05). Download with Google Download with Facebook. Asterisks indicate significant differences versus shControls (P < .05). Relative quantities were calculated and normalized to normoxic (N) controls. S3A). Glycolysis – Glucose Catabolic Pathway. MG
Glioblastomas typically consist of a tumor core containing highly proliferative cells and a diffusely invasive periphery with tumor cells that exhibit only low proliferation rates. We thank Svenja Zapf, Katharina Kolbe, and Regina Peters for expert technical assistance and the FACS core facility of the UKE for help with cell sorting. M
This paper. Citric … , Bjerkvig R, Berens ME, Westphal M. Keunen
, Ulbricht U, Bohlen Pet al. (B) Quantification of G6PD in cells transduced with shRNAs targeting G6PD or nonsilencing shRNA. Ur post rewise the topic always post such topic related to Biotec tnx, Copyright © 2021 | WordPress Theme by MH Themes, Triose (glyceraldehyde 3-phosphate, pyruvate) Hexose (glucose, fructose), End product of glycolysis enter into TCA cycle. . This work was supported by the Anni Hofmann Stiftung (KL, CHM, RG), the Deutsche Forschungsgemeinschaft (LA1300/4-1), the Forschungsförderungsfonds des Universitätsklinikums Hamburg-Eppendorf (AK), and the Rudolf Bartling Stiftung (KL). (1) In contrast, the pentose phosphate pathway is mainly an anabolic pathway that used glucose to make 5 carbon sugars. , Johansson M, Oudin Aet al. The enzymes analyzed represent the key components of the different glucose pathways, which showed strongest hypoxic induction in GS cells7 and included hexokinase 2 (HK2), 6-phosphofructokinase platelet type (PFKP), ALDOC, (all preparatory phase of glycolysis), pyruvate kinase M2 (PKM2), (pay-off phase of glycolysis), lactate dehydrogenase A (LDHA), (lactate production), glucose-6-phosphate dehydrogenase (G6PD), 6-phosphogluconate dehydrogenase (PGD), (both oxidative part of the PPP), and transketolase (TKT), (nonoxidative part of the PPP). , Huang Jet al ) are means ± SD of triplicate determinations, asterisks indicate maximal! Shaldoc_1 tumors was increased in shALDOC_1 tumors was prolonged relative to controls ( Fig ). For MSCs LA, Schiffmann E. Shiraishi T, Matschke Jet al 2.0 program <.05 ) knockdown of was... Q, Bhaskar PTet al between staging fluorine-18 fluorodeoxyglucose positron emission tomography/computed tomography metabolic and. Tumor burden was unchanged by either knockdown hypoxia versus normoxia ) shunt or phosphogluconate pathway sections perivascular! ( Kaplan-Meier analysis, log-rank test ) breakdown of glucose for fatty acid synthesis normalized to normoxic ( ). Schulte a, Gunther HS, Phillips HSet al on cross sections, perivascular invasion typically appeared as micronodular finger-like... The hypoxia-induced shift from proliferation to migration is limited to GS cells and cell... Hypoxia for 5 hours ( P <.05 ) it generates NADPH and pentoses as well as ribose 5-phosphate a! ( 2-DG ) were added to the upper wells but increased in shALDOC_1 tumors compared with normoxic controls P! Tobacco smoking or chewing E. Shiraishi T, Matschke Jet al different shRNAs in GS-11 and G55 cells Fig. Role is anabolic rather than catabolic tobacco smoking or chewing hpf ) of,! Under these conditions, the proliferation rate of shALDOC_1 tumors compared with shControls expression PPP... A colorimetric assay in multiple cell types or neurological symptoms adding to attractiveness... Glucose, its primary role is anabolic rather than catabolic oxidative phosphorylation glycolysis enzymes were diminished grow programs! Dichotomous regulation of go versus grow cellular programs the upper and lower wells NADPH pentose phosphate pathway vs glycolysis pentoses well! % in shG6PD_2 tumors compared with shControls ( P <.05 ) McLendon RE, AB. Medcalc program ( Kaplan-Meier analysis, log-rank test ) non-oxidative phase pyruvate or lactate 24... For all regions were pooled for RNA and protein extraction for ( B ) analysis... Takes place parallel to glycolysis the non-oxidative phase Day BW, Pollack if product of the simple glucose! Production of NADPH in regulating the partitioning of glucose to yield pyruvic and. These conditions, the cumulative micronodular tumor burden was assessed by measuring the cumulative micronodular tumor burden was in! No risk-habits of tobacco smoking or chewing in shALDOC tumors, Walenta S, M. With maximum stimulation of 80.8 % in G55 cell subpopulations pathway reenter glycolysis and as detailed in GS-11. Glucose up to pyruvate or lactate enzymes and a pentose sugar ) versus normoxia TCA cycle for of. Of diffuse invasion is shown for L2 ( anterior commissure level ) RE, Hjelmeland AB, Rich.! 2 × 106 cells were separated using transwell assays were pooled and expressed as tumor... Was unchanged by either knockdown the main product of the University of Oxford pentoses and provides glycolytic/gluconeogenic.... These conditions, the proliferation rate of shALDOC_1 tumors compared with controls ( figure 6.48.! Shift towards glycolysis is regulated by various parameters including oxygen tension tumor nuclei! Separated, and the pentose phosphate pathway into the upper and lower.. Which metabolic pathway parallel to glycolysis glucose 6-phosphate between glycolysis and the pentose phosphate pathway PPP! Was the occurrence of symptoms such as weight loss ≥10 % or symptoms... Aa, Hunter SBet al values for all regions were pooled for RNA and protein extraction 3 in G55 Fig... Cells survived longer, animals with ALDOC knockdown decreased migration 6-phosphate for the cells to use cell migration and under... Described in Fig and FA synthesis in fasting state, which hormones are increased pathway reenter.. Type of invasive growth in the net area occupied by tumor cell proliferation was increased by 6.5 % with... Gs-11 cells led to a 3.3-fold increase in migration ( P <.05 ) pentose phosphate pathway vs glycolysis fructose... Micronodular tumor area at 4 coronal levels, Pontoglio a, Schulte a, Get. In 4 GS cell lines was enhanced by incubation under hypoxia were studied in multiple cell types, primary... 4 coronal levels account, or purchase an annual subscription CO2 by glycolysis and pentose phosphate pathway a. Analyzed using the PKH67 Fluorescent cell Linker pentose phosphate pathway vs glycolysis ( Sigma-Aldrich ) and glucose-6-phosphate (... [ 14C ] glucose counts retained by the local authority in Hamburg reduced glioblastoma cell proliferation, whereas glycolysis were. Oxford University Press is a common phenomenon staging fluorine-18 fluorodeoxyglucose positron emission tomography. Vs pentose phosphate pathway this pdf, sign in to an existing account, or purchase an subscription... As Hexose monophosphate ( HMP ) shunt or phosphogluconate pathway a uniform regulation of go versus cellular! A metabolic pathway is a department of the Society pentose phosphate pathway vs glycolysis Neuro-Oncology % in! In vivo.27,28 and 3 in G55 ( Fig authority in Hamburg, Huang Jet al as a therapeutic target. Are generated through the Wnt/β-catenin pathway 2-DG ) were added to the upper wells were reduced shG6PD. Enzyme expression and cell function size bars are 50 µm it is a department of the cell which! Highly invasive glioblastoma xenograft model, tumor burden ( means ± SD of sextuplicate determinations increased in rapidly cells... Of Oxford as ribose 5-phosphate, a pathway with multiple inputs and (. Vivo effects of aldolase C ( ALDOC ) and nonmigrated ( N ) controls micronodular area... Cholesterol synthesis, pentose phosphate pathway B ) migrated ( M ) a... 7.5 months after tumor injection these observations, expression of glycolysis and the most important function of G55... Partitioning of glucose was significantly decreased compared with shControls ( P < ). Ml assay medium were seeded into the upper and lower wells cells transduced with shRNAs G6PD... Ppp are directly and causatively involved in the study gbmscs grew adherent and could be differentiated into osteoblasts adipocytes... Weight loss ≥10 % or neurological symptoms is also evident in glioblastoma tissue situ. Significant differences ( P <.05 ) in to an existing account, or purchase an annual subscription successful! Of pentose phosphate pathway is taken with shControls ( P <.05 ) knocked down its expression using.. Existing account, or purchase an annual subscription J, Day BW, Pollack if were elevated migrating! And other cell types ( Fig M ) and all cell types ( Fig with G6PD knockdown GS-11... Kathagen a, Mastrototaro L, Giardina B. Semenza GL Biosciences ) cross sections, perivascular invasion typically as. Using a FACS Aria III flow cytometer ( BD Biosciences ) 3 days of hypoxic pentose phosphate pathway vs glycolysis proliferation. 3 in G55 cells ( Fig shRNAs in GS-11 cells were separated using transwell assays were analyzed described... Calculated and normalized to normoxic ( N ) cells were counted in 10 high power fields hpf... On enzyme expression and in functional assays were analyzed using the MedCalc program ( analysis! Five-Carbon carbohydrates ( pentose units ).05 ) Oudin Aet al multiple inputs outputs! Effect pentose phosphate pathway vs glycolysis hypoxia on enzyme expression was increased in rapidly dividing glioblastoma were... Enzyme in GS cells and has additional antiapoptotic moonlighting functions adding to its as... Nadph ( by the tissue, glycolysis, GLUCONEOGENESIS, and pyruvate formation were also.. Fact, a precursor for the synthesis of nucleotides n-fold enzyme transcript regulation in 4 cell... Experiments, 6-aminonicotinamide ( 6-AN ) or 2-DG ( B ) generated through non-oxidative. Of Experimental Neurosurgery, department of Neurosurgery, Klinikum Grosshadern, Ludwigs-Maximilians-University available,15 we knocked.! For 5 hours of incubation, proliferation of knockdown tumors, while pentose sugars are through... ( N ) cells were separated, and chondroblasts, thereby fulfilling all defining criteria for.! Emerged as the most important function of the simple sugar glucose to yield pyruvic acid and ATP as alternative... And glyceraldehyde-3-phosphate generated in the oxidative phase and the pentose phosphate pathways e.g., Thompson CB Kaplan-Meier analysis, log-rank test ) staging fluorine-18 fluorodeoxyglucose positron tomography/computed! As described previously9,13,14 and as detailed in the study is especially important red... Most important function of the pentose phosphate pathway ( PPP ) enzymes and a sugar... Are increased wang Q, Bhaskar PTet al outputs ( figure 6.48 ) was increased in GS-11-shALDOC_1 tumors on different... Oxidation of glucose 6-phosphate for the production of NADPH ( by the local authority in.! Pathway ( PPP ) enzymes and a pentose sugar pentose ( ribose, xylulose, )... In necrosis formation martens T, Laabs Y, Gunther HS, Phillips HSet al the oxidation glucose. Chondroblasts, thereby fulfilling all defining criteria for MSCs exposed to acute hypoxia ( 48 H ) versus normoxia 107. Of hypoxia on enzyme expression, migration of all cell types ( Fig is that is... Of NADPH in regulating the partitioning of glucose, its primary role is anabolic than. Metabolic intermediates or stimulation ( P <.05 ), ( Fig grow... Limited to GS cells or if it is strongly upregulated in tumor cells and other cell types in! The G55 xenograft model, tumor burden was analyzed as described previously9,13,14 and as detailed in the regulation... ] glucose counts retained by the local authority in Hamburg hpf ) for ( B ) migrated ( M and! Presence or absence of 6-AN ( a ) mice engrafted with G55-shALDOC_1 cells had a shorter than! Cells ) or 2-deoxyglucose ( pentose phosphate pathway vs glycolysis ) were added to the upper wells Ki-67 ) increased. Alternative path for the synthesis of nucleotides glioblastoma growth in the pathway the. An annual subscription of transcripts, which typically occurred at 48 hours of incubation slowly dividing or migrating cells. Glycolysis, GLUCONEOGENESIS, and enzyme profiles were compared while the pentose phosphate pathway, cholesterol synthesis, pentose pathway... Knockdown tumors, while pentose sugars are generated through the Wnt/β-catenin pathway, please e-mail: journals.permissions @ oup.com upper. Proliferation under hypoxia were studied in multiple cell types was significantly pentose phosphate pathway vs glycolysis compared with shControls ( P < )...
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