Triclosan　(TCS),　an　antimicrobial　compound,　is　known　to　have　potential　endocrine-disruptive　properties,　but　the　underlying　toxic　mechanisms　at　the　metabolic　level　are　not　well　understood.　Here,　we　applied　metabolomics　and　lipidomics　combined　with　mass　spectrometry　imaging　(MSI)　to　unveil　the　mechanisms　of　the　enhanced　growth　of　MCF-7　breast　cancer　cell　spheroids　(CCS)　exposed　to　TCS.　To　obtain　a　wide　coverage　of　metabolites　and　lipids　by　using　MSI,　we　used　techniques　of　matrix-assisted　laser　desorption/ionization　(MALDI)　and　MALDI　coupled　with　laser-postionization.　The　results　showed　that　TCS　and　TCS　sulfate　penetrated　into　the　entire　area　at　0-3　h　and　both　localized　in　the　inner　area　at　6　h.　After　24　h,　a　portion　of　two　compounds　was　released　from　CCS.　Omic　data　indicated　that　TCS　exposure　induced　alterations　via　several　pathways,　including　energy　metabolism　and　biosynthesis　of　glycerophospholipids　and　glycerolipids.　Further　MSI　data　revealed　that　the　enhancement　of　energy　supply　in　the　peripheral　area　and　the　increase　of　energy　storage　in　the　inner　area　might　contribute　to　the　enhanced　growth　of　MCF-7　breast　CCS　exposed　to　TCS.　This　study　highlights　the　importance　of　integrating　metabolite　distributions　and　metabolic　profiles　to　reveal　the　novel　mechanisms　of　TCS-triggered　endocrine　disrupting　effects.　©　2023　The　Authors.　Published　by　American　Chemical　Society.