OBJECTIVE: The aim of this study is to investigate the expression and clinical significance of 6-phosphofructo-1-kinase in nasopharyngeal carcinoma biopsy samples. METHOD: Sixty-one biopsy samples were detected, including 41 tissues samples from patients with nasopharyngeal carcinoma and 20 tissues samples from patients with chronic nasopharyngitis as control group. 6-phosphofructo 1 kinase mRNA expression was detected by RTPCR. RESULT: It was observed that the expression levels of 6 phosphofructo-1-kinase mRNA in nasopharyngeal carcinoma tissues were higher than in the chronic inflammatory tissues. And the expression levels in patients with lymph node metastasis were higher than without lymph nod metastasis. CONCLUSION 6-phosphofructo-1-kinase may be a marker in occurrence and metastasis of nasopharyngeal carcinoma.
Biomarkers, Tumor ; metabolism ; Carcinoma ; Female ; Humans ; Lymphatic Metastasis ; Male ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Phosphofructokinase-1 ; genetics ; metabolism ; RNA, Messenger ; metabolism

OBJECTIVE: The aim of this study is to investigate the expression of phosphofructokinase 1 and it's enzyme activity in nasopharyngeal carcinoma biopsy samples. METHOD: Sixty-one biopsy samples were detected, including 41 tissues from patients with nasopharyngeal carcinoma as experimental group and 20 tissues from patients with chronic nasopharyngitis as control group. Phosphofructokinase 1 protein was detected by Western blot and it's enzyme activity was detected. RESULT: It was observed that the expression levels of phosphofructokinase 1 protein and it's enzyme activities in the experimental group were higher than that in the control group (P < 0.01). In the experimental group, the expression levels of phosphofructokinase 1 protein and it's enzyme activities in patients with lymph node metastasis were higher than that in patients without lymph node metastasis (P < 0.01). CONCLUSION Phosphofructokinase 1 may be a marker in occurrence and metastasis of nasopharyngeal carcinoma.
Biomarkers, Tumor ; metabolism ; Biopsy ; Carcinoma ; Case-Control Studies ; Humans ; Lymphatic Metastasis ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; enzymology ; pathology ; Nasopharyngitis ; Phosphofructokinase-1 ; metabolism

OBJECTIVETo investigate the influence of hypoxia induction factor-1alpha (HIF-1alpha) on glycosis of rat myocardial cell under hypoxic condition.

METHODSThe myocardial cells of the rats were routinely isolated and cultured. The cells were divided into single hypoxia (H) and HIF-1alpha inhibiting (I) groups. The cells in H group were cultured in glucose-free medium with mixed low-oxygen gas [1% O2, 94% N2 and 5% CO2 (v/v)]. While the cells in I group were cultured with low-oxygen gas after the cell model of low expression of HIF-1alpha protein constructed by RNAi technique. The cells in both groups were all observed before hypoxia (routine culture) and at the time points of 1, 3, 6, 12 and 24 hours of hypoxia. The LA (lactate acid ) content in the supernatant of the culture and the activity of the key enzymes in glycolysis such as hexokinase (HK), phosphofructokinase (PFK) and lactate dehydrogenase (LDH) of both groups of cells were determined at all the time points.

RESULTS(1) After hypoxia, the HK and PFK activities of the rat myocardial cells in H and I groups were obviously increased at the beginning and decreased thereafter when compared with that before hypoxia. While the activities of HK and PFK in H group at 1, 3 and 6 hours after hypoxia were evidently higher than those in I group (P <0.05 or 0.01), and the peak activity of them in H and I groups was 159 +/- 13 U/g vs 133 +/- 55 U/g, and 298 +/- 44 U/g vs 188 +/- 55 U/g, respectively. (2) Compared with normal control (92 +/- 12 U/g), the LDH activity of the cells in H group after hypoxia increased significantly, reaching the peak at 6 hours after hypoxia (2 568 +/- 125 U/g, P < 0. 01), and it decreased thereafter, while that in I group peaked at 3 hours after hypoxia (2125 +/- 126 U/g, P <0.01). The LA content in the culture supernatant in H group increased significantly after hypoxia with the passage of time, while that in I group increased in smaller magnitude (P <0.01).

CONCLUSIONHigh expression of HIF-1alpha in the rat myocardial cells after hypoxia could directly cause continuous enhancement of cell glycolysis, which was beneficial to the protection of myocardial cells under hypoxic condition.

Animals ; Cell Hypoxia ; Cells, Cultured ; Glycolysis ; Hexokinase ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Myocytes, Cardiac ; metabolism ; Phosphofructokinase-1 ; metabolism ; RNA Interference ; Rats ; Rats, Sprague-Dawley

After renal injury, selective damage occurs in the proximal tubules as a result of inhibition of glycolysis. The molecular mechanism of damage is not known. Poly(ADP-ribose) polymerase (PARP) activation plays a critical role of proximal tubular cell death in several renal disorders. Here, we studied the role of PARP on glycolytic flux in pig kidney proximal tubule epithelial LLC-PK1 cells using XFp extracellular flux analysis. Poly(ADP-ribosyl)ation by PARP activation was increased approximately 2-fold by incubation of the cells in 10 mM glucose for 30 minutes, but treatment with the PARP inhibitor 3-aminobenzamide (3-AB) does-dependently prevented the glucose-induced PARP activation (approximately 14.4% decrease in 0.1 mM 3-AB-treated group and 36.7% decrease in 1 mM 3-AB-treated group). Treatment with 1 mM 3-AB significantly enhanced the glucose-mediated increase in the extracellular acidification rate (61.1±4.3 mpH/min vs. 126.8±6.2 mpH/min or approximately 2-fold) compared with treatment with vehicle, indicating that PARP inhibition increases only glycolytic activity during glycolytic flux including basal glycolysis, glycolytic activity, and glycolytic capacity in kidney proximal tubule epithelial cells. Glucose increased the activities of glycolytic enzymes including hexokinase, phosphoglucose isomerase, phosphofructokinase-1, glyceraldehyde-3-phosphate dehydrogenase, enolase, and pyruvate kinase in LLC-PK1 cells. Furthermore, PARP inhibition selectively augmented the activities of hexokinase (approximately 1.4-fold over vehicle group), phosphofructokinase-1 (approximately 1.6-fold over vehicle group), and glyceraldehyde-3-phosphate dehydrogenase (approximately 2.2-fold over vehicle group). In conclusion, these data suggest that PARP activation may regulate glycolytic activity via poly(ADP-ribosyl)ation of hexokinase, phosphofructokinase-1, and glyceraldehyde-3-phosphate dehydrogenase in kidney proximal tubule epithelial cells.
Animals ; Cell Death ; Epithelial Cells* ; Glucose ; Glucose-6-Phosphate Isomerase ; Glycolysis ; Hexokinase ; Kidney* ; LLC-PK1 Cells ; Oxidoreductases ; Phosphofructokinase-1 ; Phosphopyruvate Hydratase ; Poly Adenosine Diphosphate Ribose* ; Poly(ADP-ribose) Polymerases* ; Pyruvate Kinase ; Swine

OBJECTIVETo investigate the influence of microtubule intervention drugs on glycolytic key enzymes in myocardial cells after hypoxia.

METHODSThe primary passage of cultured myocardial cells from neonatal rats were divided into A group (with hypoxia), B group (with hypoxia and administration of l0 micromol/L colchicine), C group (with hypoxia and administration of 5 micromol/L taxol), D group (with hypoxia and administration of 10 micromol/L taxol), E group (with hypoxia and administration of 15 micromol/L taxol). The morphology of microtubule was observed with laser scanning microscope (LSM). The cell vitality was assayed by cell counting kit (CCK). The activities of hexokinase (HK), pyruvate kinase (PK), phosphofructokinase (PFK) and lactate dehydrogenase (LDH) were assayed with colorimetry.

RESULTSIn group B and E, the microtubule structure was damaged heavily, and the cell vitality was decreased significantly [The cell vitality was (89.99 +/- 3.47)% in B group and (84.56 +/- 6.61)% in E group, respectively, at 1.0 post hypoxia hour (PHH), and hoth values were obviously lower than that in A group (97.44 +/- 1.76)%, P < 0.01]. The HK, PK and PFK activities decreased obviously. The activities of HK, PK and PFK in group C were similar to those of the A group. Compared with that in other groups, the degree of damage of microtubule structure in D group was milden. The activities of HK, PK and PFK in D group during 0.5 - 6.0 PHH were significantly higher than those in A group. The activity of LDH in each group was increased after hypoxia.

CONCLUSIONProper concentration of microtubule-stabilizing drugs can alleviate the damages to microtubule structure, and enhance the activity of glycolytic key enzymes of myocardial cells at early stage of hypoxia.

Animals ; Cell Hypoxia ; Cells, Cultured ; Glycolysis ; drug effects ; Hexokinase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Microtubules ; drug effects ; metabolism ; Myocytes, Cardiac ; enzymology ; metabolism ; Phosphofructokinase-1 ; metabolism ; Pyruvate Kinase ; metabolism ; Rats ; Rats, Sprague-Dawley

This research aimed to study the effect of distillage recycling on ethanol fermentation, the key glycolytic enzymes and cell composition of the self-flocculating yeast. With the self-flocculating yeast SPSC01 and medium composed of 220 g/L glucose, 8 g/L yeast extract and 6 g/L peptone, continuous ethanol fermentation was carried out at the dilution rate of 0.04 h(-1) with a 1.5 L tank bioreactor. Fermentation broth was collected every 3 days, and ethanol and other volatile byproducts were removed by distillation, but the stillage with high boiling byproducts was recycled to prepare the medium instead of fresh water. The system was run for 20 days, during which ethanol and biomass concentrations in the effluent decreased continuously, indicating the significant inhibition of the high boiling byproducts accumulated within the system. Thus, the activities of the key enzymes of the glycolytic pathway: hexokinase, 6-phosphofructose kinase, and pyruvate kinase were analyzed, and it was observed that all of them were inhibited. Furthermore, the biosynthesis of the stress response metabolites glycerol and trehalose was investigated, and it was found that glycerol production that can protect yeast cells against osmotic pressure stress was enhanced, but trehalose biosynthesis that can protect yeast cells against ethanol inhibition was not improved, correspondingly. And in the meantime, the biosynthesis of the major intracellular components proteins and hydrocarbons was adjusted, correspondingly.
Bioreactors ; microbiology ; Ethanol ; metabolism ; Fermentation ; Flocculation ; Glycerol ; metabolism ; Glycolysis ; Hexokinase ; metabolism ; Industrial Microbiology ; methods ; Phosphofructokinase-1 ; metabolism ; Saccharomyces cerevisiae ; enzymology ; genetics ; metabolism ; Schizosaccharomyces ; enzymology ; genetics ; metabolism ; Trehalose ; metabolism ; Triticum ; metabolism ; Zea mays ; metabolism