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 | ANGPTL4 | Glut-3 | PDGF-A | ANGPTL4 | Glut-3 | PDGF-A | ANGPTL4 | Glut-3 | PDGF-A | ANGPTL4 | Glut-3 | PDGF-A |
| B | 18SrRNA | HIF-1a | PDK1 | 18SrRNA | HIF-1a | PDK1 | 18SrRNA | HIF-1a | PDK1 | 18SrRNA | HIF-1a | PDK1 |
| C | BCL2 | HIF-2a | RTP | BCL2 | HIF-2a | RTP | BCL2 | HIF-2a | RTP | BCL2 | HIF-2a | RTP |
| D | CREBP | HPRT | SAG | CREBP | HPRT | SAG | CREBP | HPRT | SAG | CREBP | HPRT | SAG |
| E | CTSD | IGF-II | UCP3 | CTSD | IGF-II | UCP3 | CTSD | IGF-II | UCP3 | CTSD | IGF-II | UCP3 |
| F | EP300 | Leptin | VEGF | EP300 | Leptin | VEGF | EP300 | Leptin | VEGF | EP300 | Leptin | VEGF |
| G | EPO | MMP2 | VHL | EPO | MMP2 | VHL | EPO | MMP2 | VHL | EPO | MMP2 | VHL |
| H | Glut-1 | PAI-1 | Blank | Glut-1 | PAI-1 | Blank | Glut-1 | PAI-1 | Blank | Glut-1 | PAI-1 | Blank |
Principle:
Signosis’proprietaryCDNAplatearrayisaplate-basedhybridizationprofilinganalysisformonitoringtheexpressionofdozensofgenesthroughreversetranscriptionofmRNAintocDNA.Likearrayanalyses,totalRNAisfirstreversetranscribedintocDNAinthepresenceofbiotin-dUTPintheassay.Targetedgenesarethenspecificallycapturedontoindividualwellsonaplate,insteadofmembranes,throughapre-coatedgene-specificoligonucleotide.ThecapturedcDNAsarefurtherdetectedwithstreptavidin-HRP.Luminescenceisreportedasrelativelightunits(RLUs)onamicroplateluminometer.Theexpressionlevelofgenesisdirectlyproportionaltotheluminescentintensity.
Data:
MouseHIF-regulatedcDNAplatearrayanalysis. NIH3T3cellsweretreatedwithandwithout200uMcobaltchloridefor24hours,RNAsprepared,cDNAsynthesizedwithbiotinlabelandsubjectedtocDNAplatearrayhybridizationanddetection.
