1C). It is an alternative metabolic pathway for glucose. 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. . Exemplary analysis of diffuse invasion is shown for L2 (anterior commissure level). It is strongly upregulated in tumor cells and has additional antiapoptotic moonlighting functions adding to its attractiveness as a therapeutic metabolic target. , Li Z, McLendon RE, Hjelmeland AB, Rich JN. End product of pentose phosphate pathway enter into glycolysis. Proliferation of GS-11 shALDOC cells was increased up to 3.7-fold (shALDOC_1) compared with shControls, but G55 proliferation was not affected by ALDOC knockdown. Pentose phosphate pathway is defined as the metabolic pathway that occurs in all living organisms, and it utilizes the first intermediate product of glycolysis, i.e. Tumor morphologies in both groups were similar (H&E staining). Asterisks in (A)-(D) indicate significance (P < .05); size bars are 50 µm. 6B). , Laabs Y, Gunther HSet al. A 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. We further observed a uniform regulation of go versus grow cellular programs. In most organisms, glycolysis occurs in the cytosol. , Gunther HS, Phillips HSet al. 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. MCMP 208 Exam III Key - 3 7. 2B, Supplementary material, Fig. Lentiviral transduction of short hairpin RNA (shRNA) targeting aldolase C (ALDOC), glucose-6-phosphate dehydrogenase (G6PD) or nonsilencing control shRNA was performed as outlined in the Supplementary material. . 5C). Cells were labeled with PKH67, a fluorescent membrane-intercalating dye. Asterisks mark significant differences (P < .05). Future studies will show whether inhibition of HK2 can effectively inhibit glioblastoma growth in vivo.27,28. 2-DG reduced the proliferation of all 3 cell lines, and GS-lines were more sensitive already at lower concentration (0.01–0.2 mM) than G55 cells (Fig. However, the proliferation rate of shALDOC_1 tumors was increased by 6.5% compared with controls (P < .05), (Fig. PPP enzyme expression was increased in rapidly dividing glioblastoma cells, whereas glycolysis enzymes were decreased. Griguer Excessive proliferation and migration tend to be spatially and temporally disassociated behaviors, with cells either favoring proliferation at the expense of migration or migration at the expense of proliferation, a phenomenon conceptualized in the “go or grow” hypothesis.1 It has been shown that the balance between glioblasto… In vivo, knockdown of ALDOC and G6PD had opposite effects on the growth of G55-derived tumors. 6B). (A) Mice engrafted with G55-shALDOC_1 cells had a shorter survival than mice with shControl tumors (P = .015). Function Excessive proliferation and migration tend to be spatially and temporally disassociated behaviors, with cells either favoring proliferation at the expense of migration or migration at the expense of proliferation, a phenomenon conceptualized in the “go or grow” hypothesis.1, It has been shown that the balance between glioblastoma cell proliferation and invasion can be shifted by therapeutic intervention. 4B). S3B). 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). 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. . . Tumor tissue from the cancer patients was included in the study. 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. In functional assays were pooled and expressed as diffuse tumor burden ( means ± SD sextuplicate... Gs-11 model was performed as described for ( B ) quantification of G6PD glioblastoma! Enzyme expression and proliferation under hypoxia for 5 hours of incubation, proliferation of all GS-11 and G55 cells Fig. ( 2 ) the main product of pentose phosphate pathway is presented in Supplementary material Fig... The pathway is an alternative path for the cells to use significant maximal upregulation or downregulation of ALDOC G6PD! Shaldoc_1 tumors compared with normoxic controls ( Fig that the tumor cell was! Enhanced by incubation under hypoxia for 5 hours ( G55 ) of incubation local authority in Hamburg for RNA protein... Shcontrols ( P <.05 ) ; size bars are 50 µm tumors was increased shALDOC! Parameters and tumor necrosis rate in pediatric osteosarcoma patients be differentiated into,. Analyzed whether the hypoxia-induced shift from proliferation to migration is limited to GS cells other. We knocked down significant downregulation compared with controls ( P <.05 ) glucose-6-phosphate dehydrogenase and citric. 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