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Medchemexpress/DHEA(Synonyms: trans-Dehydroandrosterone; Prasterone; Dehydroisoandrosterone; Dehydroepiandrosterone)/HY-14650
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Medchemexpress/DHEA(Synonyms: trans-Dehydroandrosterone; Prasterone; Dehydroisoandrosterone; Dehydroepiandrosterone)/HY-14650
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MedChemExp
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HY-14650-10mM*1mLinDMSO
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DHEAisanimportantsourceofandrogens,andisaneffectiveantiapoptoticfactor.
Description

DHEAisanimportantsourceofandrogens,andisaneffectiveantiapoptoticfactor.

IC50&Target

Androgenreceptor[1]

InVitro

DHEAisaneffectiveantiapoptoticfactor,reversingtheserumdeprivation-inducedapoptosisinprostatecancercells(DU145andLNCaPcelllines)aswellasincoloncancercells(Caco2cellline).DHEAsignificantlyreducesserumdeprivation-inducedapoptosisinall3cancercelltypes,quantitatedwiththeAPOPercentageassay(apoptosisisreducedfrom0.587±0.053to0.142±0.0016or0.059±0.002aftertreatmentfor12hourswithDHEAorNGF,respectively;n=3,P<0.01), and="" by="" flow="" cytometry="" analysis="" (facs)="" for="" du145="" cells.="" the="" antiapoptotic="" effect="" of="" dhea="" is="" dose="" dependent="" with="" an="" ec50="" at="" nanomolar="" concentrations="">50:11.2±3.6nMand12.4±2.2nMinDU145andCaco2cells,respectively)[1].DHEAistheprincipalsexsteroidprecursorinhumansandcanbeconverteddirectlytoandrogens.DHEA(≥1μM)causesadose-dependentinhibitionofChub-S7proliferation,asassessedbythymidineincorporationassays.DHEAtreatmentinhibitsexpressionofthekeyglucocorticoid-regulatinggenesH6PDH(≥100nM)andHSD11B1(≥1μM)indifferentiatingpreADIpocytesinadose-dependentmanner.Inkeepingwiththisfinding,DHEAtreatment(≥1μM)resultsinamarkedreductionin11β-HSD1oxoreductaseactivity(≥1μM)andaconcurrentincreaseindehydrogenaseactivityatthehighestDHEAdoseused(25μMDHEA)indifferentiatedadipocytes[2].

InVivo

DHEAinthediet(0.45%w/w)ofmaleB6mice(groupsoffivemice)treatedfor8weeksledtosignificantdecreasesinbodytemperaturecomparedwithmicefedthecontrolAIN-76Adiet.Asimilarcomparisonindicatedthatcontrolandpair-fedmicearealsosignificantlydifferent.AnimalsfedDHEAhavesignificantlylowertemperaturesthanmicefedthecontroldiet26/29timestested;micepairfedtothoseontheDHEAdietarelessaffected,with8/29valuessignificantlylowerthaninmicefedAIN-76Aadlibitum.ThetemperaturesofmicefedDHEAorpairfedtoDHEAaresignificantlydifferent21/29timestested.BodyweightsaresignificantlygreaterinmicefedthecontroldietthaninmicefedDHEAorpairfedtoDHEA.Foodintake(gramsperday)fromcagesareaveragedforeachweek(n=7),exceptforWeek9(n=3).TheamountoffoodintakeissignificantlydecreasedinmicefedDHEA.Bydesign,micepairfedtoDHEAateaboutthesameamount.Thus,itappearsthatDHEAreducesbodytemperaturebyfoodrestrictionandbyaseparatemechanism[3].

References

  • [1].AnagnostopoulouV,etal.Differentialeffectsofdehydroepiandrosteroneandtestosteroneinprostateandcoloncancercellapoptosis:theroleofnervegrowthfactor(NGF)receptors.Endocrinology.2013Jul;154(7):2446-56.

    [2].McNelisJC,etal.Dehydroepiandrosteroneexertsanti-glucocorticoidactiononhumanpreadipocyteproliferation,differentiationandglucoseuptake.AmJPhysiolEndocrinolMetab.2013Nov1;305(9):E1134-44.

    [3].CatalinaF,etal.Decreaseofcorebodytemperatureinmicebydehydroepiandrosterone.ExpBiolMed(Maywood).2002Jun;227(6):382-8.

PreparingStockSolutions
ConcentrationVolumeMass1mg5mg10mg
1mM3.4672mL17.3358mL34.6717mL
5mM0.6934mL3.4672mL6.9343mL
10mM0.3467mL1.7336mL3.4672mL
Pleaserefertothesolubilityinformationtoselecttheappropriatesolvent.
KinaseAssay
[2]

Chub-S7cellsareincubatedinDMEMcontainingcoldDHEA(20nM)andtritiatedDHEA(0.2μCi/well)for48h.Followingincubation,steroidsareextractedusingdichloromethaneseparatedbythin-layerchromatographyusingn-hexane/1-hexanol(75:25)asthemobilephasesystem.MetabolitesareidentifiedbycomigrationwithunlabeledreferencesteroidsthatarevisualizedbyexposuretoLieberman-Burchardreagent(ethanol-aceticanhydride-sulfuricacid).SteroidconversionisquantifiedusingaLabLogicAR-200scanner.Proteinconcentrationismeasuredusingacolorimetric96-wellplateassayandusedtonormalizeconversion.Activityisexpressedaspercentconversion[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

CellAssay
[2]

DHEAisdissolvedinDMSOandstored,andthendilutedwithappropriatemediumbeforeuse[2].

Chub-S7preadipocytesandhumanprimarypreadipocytesareseededintoa24-wellplateatdensities1×105and2.5×105respectively.Followingovernightculture,mediumissupplementedwithDHEA,androstenediol,orDHEAS(0-100μM).Following24-,48-,or72hincubation,cellproliferationisassessedbyincubationwithradiolabeledthymidine(0.2μCi/well)forthefinal6hofculture.ProteinsareprecipitatedwithTCA,andcellsarescrapedinNaOH.Therespectivecontentofradiolabelednuclearmaterialintheresultinglysatesisanalyzedbyscintillationcounting.Dataareexpressedaspercentageofcontrol[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

AnimalAdmiNISTration
[3]

DHEAispreparedin0.9%NaCl(Mice)[3].

Mice[3]
MicearefedPurinaLabChowuntilthestartofexperiments(Day0).GroupsoffivemicearethenfedpelletedAIN-76AdietcontainingeithernoadditiveorDHEA(0.45%w/w)between0900and1000hr.Dietsarestoredat4°Cfornolongerthansixmonthstomaintainoptimalactivity.Micearegiventhedietsadlibitum,exceptformicethatarepairfedtomicetreatedwithDHEA.TheamountsofAIN-76Adietthepair-fedmicereceivedaredeterminedbytheweightoffoodconsumedbytheDHEA-fedmiceonadailybasis.Bodyweights(grams)aremeasuredatdifferenttimepointsstartingatDay1andendingatDay59.Dailyfoodintakes(gramsperday)aredeterminedbyweighingthefoodconsumedpercageoffivemice.Themean±SEMvaluesarecalculatedforweeks1to8(n=7);week9hadonly3days.MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.

References

  • [1].AnagnostopoulouV,etal.Differentialeffectsofdehydroepiandrosteroneandtestosteroneinprostateandcoloncancercellapoptosis:theroleofnervegrowthfactor(NGF)receptors.Endocrinology.2013Jul;154(7):2446-56.

    [2].McNelisJC,etal.Dehydroepiandrosteroneexertsanti-glucocorticoidactiononhumanpreadipocyteproliferation,differentiationandglucoseuptake.AmJPhysiolEndocrinolMetab.2013Nov1;305(9):E1134-44.

    [3].CatalinaF,etal.Decreaseofcorebodytemperatureinmicebydehydroepiandrosterone.ExpBiolMed(Maywood).2002Jun;227(6):382-8.

MolecularWeight

288.42

Formula

C₁₉H₂₈O₂

CASNo.

53-43-0

Storage
Powder-20°C3years
 4°C2years
Insolvent-80°C6months
 -20°C1month
Shipping

RoomtemperatureincontinentalUS;mayvaryelsewhere

Solvent&Solubility

10mMinDMSO

*"<1 mg/ml"="" means="" slightly="" soluble="" or="" insoluble.="" "≥"="" means="" soluble,="" but="" saturation="">

Purity:>98.0%

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