LC-MS/MS Analysis of Renal Cell Carcinoma Treated with Sulforaphane

Sulforaphane(SFN)isanisothiocyanate(ITC)derivedfromcruciferousvegetables,suchas
broccoli,thatisgrowinginpopularityforitsantioxidantandanti-inflammatorycapabilities.
Furthermore,SFNhasbeendemonstratedtoimproverenalcancercarcinoma(RCC)treatment
outcomesinconjunctionwithmultipleotherformsoftherapy,whichisespeciallyimportant
consideringRCC’spoortherapeuticoutcomeswithchemotherapy.Theaimofthisstudywasto
determinetheeffectsofSFNonRCC ​invitro utilizingcellviabilityanalysisandLC/MS-MS
targetedmetabolicprofilingtorevealpathwaysresponsibleforSFN’spossibleenhancementof
chemotherapytreatmentinRCC.CCK-8resultsshowthat15 ​μ​MofSFNcausedasignificant(p
<0.05)increaseinRCCproliferation.Kruskal-Wallistestsrevealed16metabolitesinourcell,
and28inthemediumtobesignificant(p<0.05).Anorthogonalpartialleastsquares-discriminant
analysis,OPLS-DA,ofsignificantmetaboliteswasusedtocomparedtreatedandnon-treated
samplesforbothdatasetsandshoweda100%predictiveaccuracy(AUC=1).Enrichment
analysisdeterminedthatatotalof7metabolicpathwaysweresignificantlyenriched(VLCFA
β-oxidation,glutamatemetabolism,theureacycle,ammoniarecycling,glycine/serine,alanine,
andglucose-alaninecycle).Pathwayanalysisshowedhistidinemetabolismtobetheonly
significantlyaffectedpathwaybetweenbothdatasets.SFN-inducedmetaboliccharacteristics
foundinRCCwereconsistentwithknownantioxidantandanti-inflammatorypathways.Ourdata
suggeststhatthetherapeuticmechanismsofSFNarelikelyduetointeractionswithTandNKT
cellsthatprotectthemfromoxidativestress.Futureexperimentsregardingantioxidantresearch
incancershouldbecompletely ​invivo​,asopposedto ​invitro, ​inordertomaintainthenatural
physiology of cancer cells in the presence of host immune cells.