Description:
IL-6ResponsiveLuciferaseReporterHEK293CellLineisderivedhumanembryonickidney,andstablyexpressfireflyluciferasereportergeneunderthecontrolofIL-6responseelement.ThiscelllineisanidealcellularmodelformonitoringtheactivationofMAPKs,PI3Ks,STATsresponsePathwaytriggeredbystimulitreatment,enforcedgeneexpressionandgeneknockdown.Principle:
Interleukin-6(IL-6)isamultifunctionalcytokinewhichplaysanimportantroleinawiderangeofBIOLOGicprocessesindifferenttypesofcellincludingcancercells.IL6canstimulatemultiplepathwaysinvolvingMAPKs,PI3Ks,STATs,andothersignalingproteins,andsubsequentlyactivateitsdownstreamtargetsintheregulationofcellproliferation,survival,andmetabolism.EventhoughIL6wasoriginallycharacterizedasaregulatorofimmuneandinflammatoryresponses,elevatedexpressionandactivitiesofIL-6hasbeendetectedinmultiplecancers,suchasbreast,lungandlymphoma. IL-6wasassociatedwithtumorprogressionthroughincreasingantiapoptoticactivity,stimulationofangiogenesis,anddrugresistance.TheincreasedserumIL-6concentrationsinpatientsarelinkedtotheadvancedtumorstagesofvariouscancersandshortsurvivalrate.DevelopingdrugstoblockpathologicalIL-6overproductionhasbecomeapotentialtherapeuticstrategyforcancers.
PrinciplebehindTFluciferasereporter. TFluciferasereporterstablecelllineutilizesartificialpromoterconstructstodriveluciferaseexpression. Thepromoterregioncanconsistsofmultiplerepeatsofacis-elementTFbindingsite,aDNAfragmentfromthepromoterregionofaknownTFdownstreamgene,oraDNAfragmentcontainingputative/knownTFbindingsites. ThereareseveralwaysthataTFcanbeactivated,suchasthroughextracellularstimuliorthroughintracellularsignalingpathways. Onceactivated,theTFtranslocatestothenucleusandofteninteractswithrelevantco-factorstodrivegeneexpression. Onceluciferaseisexpressed,itcangeneratelightinanenzymaticassayandtheamountoflightmeasuredispositivelycorrelatedwiththelevelofTFactivation. |
Data:
AnalysisofSL-0048IL-6reporteractivityinresponsetoDifferntcytokinetreatment. Thecellswereseededona96-wellplatein10%FBSDMEDmedium.Thecellsthenimmediatelyweretreatedwithorwithout10ng/mlIL-6,IL-15,IL-8,IFNr,IL-6TNFαinDMEMand10%FBSfor24hours.IL-6 inducedover40-foldincreaseinluciferaseactivitycomparedtountreatedcells.
