Characterization
Description(s):Rapidly growing tumors result in hypoxic regions. Adaptive responses of most cells to hypoxia are (1) to produce VEGF and other hypoxia-induced angiogenic cytokines that promote increased tissue vascularization, thereby increasing tissue oxygenation, and (2) to switch metabolically from oxidative phosphorylation to anaerobic glycolysis. Hypoxia-inducible factor 1 (HIF-1) is an oxygen-regulated transcriptional activator that plays essential roles in the process. The HIF-1alpha subunit is oxygen-dependent ubiquitination and proteasomal degradation. In addition, cytokines including interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) stimulate HIF-1 dependent gene expression. HIF-1 increases the expression of several genes that promote blood flow and inflammation, such as vascular endothelial growth factor (VEGF).
Signosis has developed a plate-based array for profiling 20+ HIF-regulated genes. By using this assay, researchers are able to compare gene expression in up to three samples on a single microtiter plate.
Applicable Grid:
List of Applicable Genes
| 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | |
| A | BCL2 | HIF-2a | PDGF-A | BCL2 | HIF-2a | PDGF-A | BCL2 | HIF-2a | PDGF-A | BCL2 | HIF-2a | PDGF-A |
| B | Beta-actin | HPRT | PDK1 | Beta-actin | HPRT | PDK1 | Beta-actin | HPRT | PDK1 | Beta-actin | HPRT | PDK1 |
| C | CA9 | IGF-II | NDRG1 | CA9 | IGF-II | NDRG1 | CA9 | IGF-II | NDRG1 | CA9 | IGF-II | NDRG1 |
| D | CTSD | Leptin | SAG | CTSD | Leptin | SAG | CTSD | Leptin | SAG | CTSD | Leptin | SAG |
| E | BHLHB2 | MMP1 | SLC6A8 | BHLHB2 | MMP1 | SLC6A8 | BHLHB2 | MMP1 | SLC6A8 | BHLHB2 | MMP1 | SLC6A8 |
| F | Glut-1 | MXI-1 | VEGF | Glut-1 | MXI-1 | VEGF | Glut-1 | MXI-1 | VEGF | Glut-1 | MXI-1 | VEGF |
| G | Glut-3 | NDRG2 | VHL | Glut-3 | NDRG2 | VHL | Glut-3 | NDRG2 | VHL | Glut-3 | NDRG2 | VHL |
| H | HIF-1a | PAI-1 | Blank | HIF-1a | PAI-1 | Blank | HIF-1a | PAI-1 | Blank | HIF-1a | PAI-1 | Blank |
Principle
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Signosis’ proprietary cDNA plate array is a plate-based hybridization profiling analysis for monitoring the expression of dozens of genes through reverse transcription of mRNA into cDNA. Like array analyses, total RNA is first reverse transcribed into cDNA in the presence of biotin-dUTP in the assay. Targeted genes are then specifically captured onto individual wells on a plate, instead of membranes, through a pre-coated gene-specific oligonucleotide. The captured cDNAs are further detected with streptavidin-HRP. Luminescence is reported as relative light units (RLUs) on a microplate luminometer. The expression level of genes is directly proportional to the luminescent intensity. |
Data
Human HIF-regulated cDNA plate array analysis. HeLa cells were treated with and without 200 uM cobalt chloride for 24 hours, RNAs prepared, cDNA synthesized with biotin label and subjected to cDNA plate array hybridization and detection.
Literature
View user manualCitations
1. Endogenous Human CaMKII Inhibitory Protein Suppresses Tumor Growth by Inducing Cell Cycle Arrest and Apoptosis through Down-regulation of the Phosphatidylinositide 3-Kinase/Akt/HDM2 Pathway. Ma S, Yang Y, Wang C, Hui N, Gu L, Zhong H, Cai Z, Wang Q, Zhang Q, Li N, Cao X. J Biol Chem. 2009 Sep 11;284(37):24773-82.
2. Embryonic Overexpression of Receptors for Advanced Glycation End-Products by Alveolar Epithelium Induces an Imbalance between Proliferation and Apoptosis. Stogsdill JA1, Stogsdill MP, Porter JL, Hancock JM, Robinson AB, Reynolds PR. Am J Respir Cell Mol Biol Vol 47, Iss. 1, pp 60–66, Jul 2012
3. Δ-9,11 modifications of glucocorticoids dissociate NF-κB inhibitory efficacy from GRE-associated side effects. Baudy AR, Reeves E, Damsker J, Heier C, Garvin L, Dillingham BC, McCall J, Rayavarapu S, Wang Z, Vander Meulen J, Sali A, Jahnke V, Duguez S, Dubois D,Rose M, Nagaraju K, Hoffman E. J Pharmacol Exp Ther. 2012 Jul 3.
4. Stem cells from foreign body granulation tissue accelerate recovery from acute kidney injury. Patel J, Pancholi N, Gudehithlu KP, Sethupathi P, Hart PD, Dunea G, Arruda JA, Singh AK. Nephrol Dial Transplant. 2012 May;27(5):1780-6.
5. A novel vascular EET synthase: role of CYP2C7 Sun D, Jiang H, Wu H, Yang Y, Kaley G, Huang A. Am J Physiol Regul Integr Comp Physiol. 2011 Dec;301(6):R1723-30.
6. siRNA-mediated knockdown of aryl hydrocarbon receptor nuclear translocator 2 affects hypoxia-inducible factor-1 regulatory signaling and metabolism in human breast cancer cells. Edited by Judit Ovádi. Qin XY, Wei F, Yoshinaga J, Yonemoto J, Tanokura M, Sone H. FEBS Lett. 2011 Oct 20;585(20):3310-5.
7. Tasquinimod is an allosteric modulator of HDAC4 survival signaling within the compromised cancer microenvironment. Isaacs JT, Antony L, Dalrymple S, Brennen WN, Gerber S, Hammers H, Wissing MD, Kachhap SK, Luo J, Xing L,Bjork P, Olsson A, Björk A, Leanderson T. Cancer Res. 2012 Nov 13.
8. HIF-stabilizing bioactive glasses for directing MSC behaviour . MM Azevedo, O Tsigkou, R Nair, G Jell, J Jones… - Tissue Engineering, 2014
9.Nitration of the Tumor Suppressor Protein p53 at Tyrosine 327 Promotes p53 Oligomerization and Activation. Yakovlev VA, Bayden AS, Graves PR, Kellogg GE, Mikkelsen RB. Biochemistry., 2010 Jun 29;49(25):5331-9. doi: 10.1021/bi100564w.
10. Expression of Wnt3 activates Wnt/β-catenin pathway and promotes EMT-like phenotype intrastuzumab resistant HER2-overexpressing breast cancer cells. Vadgama JV, Wu Y, Ginther C, Kim J, Mosher N, Chung S, Slamon D. Mol Cancer Res. 2012 Oct 15.
11. A83-01 inhibits TGF-β-induced upregulation of Wnt3 and epithelial to mesenchymal transition in HER2-overexpressing breast cancer cells. Yanyuan Wu, Trinh Tran, Sami Dwabe, Marianna Sarkissyan, Juri Kim, Miguel Nava, Sheilah Clayton, Richard Pietras, Robin Farias-Eisner, Jaydutt V. Vadgama. Breast Cancer Research and Treatment. 2017 March 23. doi: 10.1007/s10549-017-4211-y
12. Conditionally induced RAGE expression by proximal airway epithelial cells in transgenic mice causes lung inflammation.B Garrett Bodine, Brock G Bennion, Emma Leatham, Felix R Jimenez, Alex J Wright, Zac R Jergensen,Connor J Erickson, Cameron M Jones, Jeff P Johnson, Steven M Knapp and Paul R Reynolds . Respiratory Research, 2014 (Published online)
13. Ablation of Doublecortin-Like Kinase 1 in the Colonic Epithelium Exacerbates Dextran Sulfate Sodium-Induced Colitis. D Qu, N Weygant, R May, P Chandrakesan. Plos One Published: August 18, 2015, DOI: 10.1371/journal.pone.0134212
