Description:
Rapidlygrowingtumorsresultinhypoxicregions.Adaptiveresponsesofmostcellstohypoxiaare(1)toproduceVEGFandotherhypoxia-inducedangiogeniccytokinesthatpromoteincreasedtissuevascularization,therebyincreasingtissueoxygenation,and(2)toswitchmetabolicallyfromoxidativephosphorylationtoanaerobicglycolysis.Hypoxia-inducIBLefactor1(HIF-1)isanoxygen-regulatedtranscriptionalactivatorthatplaysessentialrolesintheprocess.TheHIF-1alphasubunitisoxygen-dependentubiquitinationandproteasomaldegradation.Inaddition,cytokinesincludinginterleukin-1(IL-1)andtumornecrosisfactor-alpha(TNF-alpha)stimulateHIF-1dependentgeneexpression.HIF-1increasestheexpressionofseveralgenesthatpromotebloodflowandinflammation,suchasvascularendothelialgrowthfactor(VEGF).ApplicableGrid:
ListofApplicableGenes
| 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:
Signosis’proprietaryCDNAplatearrayisaplate-basedhybridizationprofilinganalysisformonitoringtheexpressionofdozensofgenesthroughreversetranscriptionofmRNAintocDNA.Likearrayanalyses,totalRNAisfirstreversetranscribedintocDNAinthepresenceofbiotin-dUTPintheassay.Targetedgenesarethenspecificallycapturedontoindividualwellsonaplate,insteadofmembranes,throughapre-coatedgene-specificoligonucleotide.ThecapturedcDNAsarefurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.Theexpressionlevelofgenesisdirectlyproportionaltotheluminescentintensity. |
Data:
HumanHIF-regulatedcDNAplatearrayanalysis. HeLacellsweretreatedwithandwithout200uMcobaltchloridefor24hours,RNAsprepared,cDNAsynthesizedwithbiotinlabelandsubjectedtocDNAplatearrayhybridizationanddetection.
