adenylate kinase glycolysis

Cells were plated in 6-well plates and treated with cycloheximide (CHX) at a concentration of 50 μg/mL for 24 h. Cells were then exposed to hypoxia for 6 h to stabilize HIF-1α protein and then switched to normoxic conditions. Cells were suspended in serum-free medium containing 0.1% bovine serum albumin and loaded into each well of the upper chamber. Inhibition of PFKFB3 cooperates with cisplatin to promote cancer cell apoptosis. The biology of cancer: metabolic reprogramming fuels cell growth and proliferation. The following antibodies were used in western blot analyses: anti-AK4 (Genetex, 1:2000), anti-HIF-1α (Cell Signaling, 1:1000), anti-hydroxylated HIF-1α (Cell Signaling, 1:1000), anti-E-cadherin (BD Bioscience, 1:1000), anti-vimentin (Sigma, 1:2000), anti-Snail (Cell Signaling, 1:1000), and anti-α-tubulin (Sigma-Aldrich, 1:5000) antibodies. The activation or inhibition status of upstream regulators in the AK4 metabolic gene signature was predicted using IPA Upstream Regulator Analysis (Ingenuity Systems, http://www.ingenuity.com), and the calculated z scores can reflect the overall activation state of the regulator (< 0: inhibited, > 0: activated). Terms and Conditions, a NOD scid Gamma (NSG) mice were injected subcutaneously with CL1-0 Vec and CL1-0 AK4 cells (1 × 106 cells/100 μL) in the left and right flanks, respectively. Lentiviruses were generated by transfecting 293 T cells with the shRNA-expression vector and pMD2.G and pDeltaR8.9 using the calcium phosphate precipitation method. Get the latest public health information from CDC: https://www.coronavirus.gov. These findings represent a novel vicious cycle between AK4 and HIF-1α in response to hypoxic stress during lung cancer progression and highlight the therapeutic opportunity of targeting the AK4-HIF-1α axis in NSCLC. Hypoxia-inducible factor-1α (HIF-1α) is a key transcription factor in the cell response to hypoxic stress. Oncogene. is a co-founder of Vivace Therapeutics. Furthermore, microarray analysis revealed that genes encoding enzymes in the glutathione metabolism pathway were differentially expressed upon AK4 overexpression in CL1-0 cells (Fig. The following antibodies were used to detect AK4, HIF-1α, E-cadherin, and pimonidazole in tissues: AK4 (Genetex, 1:200), HIF-1α (Cell Signaling, 1:100), E-cadherin (Cell Signaling, 1:100), and pimonidazole (Hypoxyprobe, INC). Swinson DE, Jones JL, Cox G, Richardson D, Harris AL, O'Byrne KJ. Low Glucose: phosphorylation ----> Privacy *P ≤ 0.05; **P ≤ 0.01. Manage cookies/Do not sell my data we use in the preference centre. R35 CA196878/CA/NCI NIH HHS/United States, Warburg O. both irreversible, Glucose + ATP ----> Glucose-6-phosphate Intriguingly, among the four members of PFKFB family, PFKFB3 is uniquely localized in the nucleus, although the reason remains unclear. Quantification of hypoxic and non-hypoxic areas in pimonidazole-stained slides was performed using Definiens’ Tissue Studio software (Definiens Inc.). 6d, e). During intense exercise: large conversion of ATP to ADP with the muscle attempting to phosphorylate ADP back to ATP, this increase in ADP pushes the reaction to the left (mass action effect) Chang X, Ravi R, Pham V, Bedi A, Chatterjee A, Sidransky D. Adenylate kinase 3 sensitizes cells to cigarette smoke condensate vapor induced cisplatin resistance. rise in AMP increases metabolism Pinho SS, Reis CA. COVID-19 is an emerging, rapidly evolving situation. Right, pimonidazole-positive tumor area was detected and quantified by Definiens imaging analysis algorithm. a 20% decrease in ATP leads to an almost 3-fold increase in ADP and ultimately a 1,400% increase in AMP. Briefly, cells were seeded in a 96-well plate (2000 cells/well) and washed with PBS. Cheng A, Zhang P, Wang B, Yang D, Duan X, Jiang Y, Xu T, Jiang Y, Shi J, Ding C, Wu G, Sang Z, Wu Q, Wang H, Wu M, Zhang Z, Pan X, Pan YY, Gao P, Zhang H, Zhou CZ, Guo J, Yang Z. Nat Commun. 2006;98(10):1254–63. PubMed  2011;6(6):e20806. glycolysis (-), Pyruvate Kinase: CL1-0 vector- and AK4-expressing cells were subcutaneously injected into NSG mice. After applying a Pearson correlation coefficient of ± 0.3 as a threshold, the AK4 metabolic gene signature was identified by selecting genes with enzyme or transporter annotations. Prevents build-up of glycolytic intermediates and the unnecessary All observations were confirmed in at least three independent experiments. 4e). The associations among AK4 IHC expression, HIF-1α IHC expression, and clinicopathological characteristics including age, gender, smoking history, tumor histology, TNM stage, pathological stage, and tumor recurrence status are summarized in Additional file 1: Table S1. PubMed Central  Other EMT regulators, such as Zeb1, Zeb2, and TCF-3, have been reported to be upregulated in pVHL-null renal cell carcinoma in which HIF-1α is constitutively overexpressed [42]. Similar to our findings, Hahm et al. *P ≤ 0.05; **P ≤ 0.01. -, Visvader J. E. & Lindeman G. J. Figure 7. hCINAP depends on its adenylate kinase activity to promote CRCs growth and invasion. Nat Rev Cancer. Cancer 8, 755–768 (2008). 1997;51(2):514–26. Cells were grown in a 6-well plate overnight, and the medium was refreshed with complete medium. Scale bar represents 2 mm. Chi-square analysis showed high expression of AK4 and HIF-1α was significantly associated lymph node involvement (Additional file 1: Table S1). Activated (+): by acetyl CoA (diverts These data are consistent with the notion that only cancer cells equipped with an enhanced antioxidant defense system are capable of leveraging oxidative stress to promote metastasis. 6-Phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 and 4: A pair of valves for fine-tuning of glucose metabolism in human cancer. 1b). 5d). The raw intensities of AK4 overexpression in CL1-0 cells (GSE37903) and lung adenocarcinoma patient datasets (GSE31210) were normalized by robust multichip analysis (RMA) using GeneSpring GX11 (Agilent Technologies). Orthotopic xenograft mouse models were established to evaluate the therapeutic efficacy of drug candidates for metastatic lung cancer. J Hematol Oncol 12, 12 (2019). Nucleoside diphosphate (NDP) kinase catalyzes in vivo ATP-dependent synthesis of ribo- and deoxyribonucleoside triphosphates. However, some studies reported inconsistent results showing that the predictive power of HIF-1α as a prognosis marker is only marginal [30,31,32]. Lee GW, Go SI, Cho YJ, Jeong YY, Kim HC, Duk Lee J, et al. 2006;66(5):2725–31. HHS Feedback inhibited by its product glucose-6-phosphate. 2011;72(1):100–7. 2006;312(10):1727–44. Figure 1. hCINAP promotes invasiveness and metastasis in CRC cells. Metabolic alterations in mammary cancer prevention by withaferin a in a clinically relevant mouse model. b Invasion assay of A549 and CL1-5 cells treated with the corresponding IC10 doses of drug candidates. The paraffin non-small cell lung cancer tissues used to generate tissue microarrays were collected from Kaohsiung Medical University Hospital with IRB approval (KMUHIRB-E(I)-20,160,099). Arteriosclerosis, Thrombosis, and Vascular Biology (ATVB), Journal of the American Heart Association (JAHA), Customer Service and Ordering Information, Basic, Translational, and Clinical Research, Adenylate Kinase–Catalyzed Phosphotransfer in the Myocardium, Cardiac Energetics in Patients With Aortic Stenosis and Preserved Versus Reduced Ejection Fraction. For scoring, both intensity and percentage of protein expression were recorded. AK4 shifts metabolism toward aerobic glycolysis and increases oxidative stress. Curr Cancer Drug Targets. In the AK4 metabolic gene signature, we identified genes encode for enzymes in N-glycan, mucin type O-glycan, and glycosaminoglycan biosynthesis pathways were significantly enriched (Fig. �$Y�6]�%�,�����y{���d�n���N�{�� �(�-��p��6�lw��}�f�\uꕏ��/��q���~ ��u��K��C�� ��m��~S�w)d s�2�/��WA�:�_B67/���謖�Gm4�]������A� �B[�y������7��{��{���y��П�β[ ���~���g4ă>I��}1ߘr 3d). Direct regulation of TWIST by HIF-1alpha promotes metastasis. 2012;72(20):5285–95. Wang J, Wang H, Liu A, Fang C, Hao J, Wang Z. Oncotarget. Clin Chim Acta. Adenylate kinase 4 modulates oxidative stress and stabilizes HIF-1α to drive lung adenocarcinoma metastasis. Science. 6f). In conclusion, we suggest that overexpression of AK4 stabilizes HIF-1α protein by increasing intracellular ROS levels and induces EMT in NSCLC. Right panel, heatmap illustrates HIF-1 α -regulated genes that are positively or negatively correlated with AK4 expression in consensus AK4 metabolic signature. on glycolysis rate [5,8] nucleotide labelling [9] and adenylate kinase activity in subcellular fractions of previously incubated isolated rat diaphragm. Taken together, our results suggest that AK4 may serve as a critical factor dictating the prognostic power of HIF-1α in lung cancer patients. Finley LW, Carracedo A, Lee J, Souza A, Egia A, Zhang J, et al. d Gross view (formalin-fixed) and H&E staining images of lungs from mice treated with DMSO vehicle control or withaferin-A (1.0 mg/kg or 4.0 mg/kg) at day 30 after orthotopic injection of CL1-0 cells overexpressing AK4 (top). Statistical analyses were performed using SPSS 17.0 software (SPSS, USA). © American Heart Association, Inc. All rights reserved. Lower glycolysis carries a higher flux than any biochemically possible alternative Steven J. Acetylation mimetic mutant of PFKFB3 disrupts its NLS motif. 6b). 6) MTT assay cell viability assay of digitoxigenin, lanatoside C, digoxin, proscillaridin, and withaferin-A in CL1-0, CL1-5, CL1-0 Vec, and CL1-0 AK4. PubMed  Adenylate kinase or ATP: AMP-phosphotrans- ferase (EC 2.7.4.3) ; apyrase or ATP diphosphohydrolase (EC 3.6.1.5); a-glucanphosphorylase or cr-1,4-glucan: ortho- phosphate-glucosyltransferase (EC 2.4.1.1), hexckinase or Adenylyl cyclase (EC 4.6.1.1, also commonly known as adenyl cyclase and adenylate cyclase, abbreviated AC) is an enzyme with key regulatory roles in essentially all cells. Figure 5. Jan, Y., Lai, T., Yang, C. et al. The authors declare no competing financial interests. The C-terminal domain of LDHA recruits hCINAP to facilitate LDHA Y10 phosphorylation in…, Figure 6. The 6-phosphofructo-2-kinase/fructose-2,6-biphosphatase 3 (PFKFB3) catalyzes the generation of fructose-2,6-bisphosphate, a potent allosteric stimulator of glycolysis.

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