Dibutyl　phthalate　(DBP),　extensively　used　as　a　plasticizer,　has　endocrine-disrupting　properties　that　may　increase　the　risk　of　breast　cancer　at　low　concentrations.　However,　previous　studies　on　the　effects　of　DBP　on　breast　cancer　mainly　focused　on　the　activation　of　typical　intracellular　receptors,　and　the　downstream　mechanisms　at　the　metabolic　level　have　not　been　well　elucidated.　Therefore,　our　study　applied　metabolomics,　lipidomics,　and　mass　spectrometry　imaging　(MSI)　techniques　to　investigate　the　metabolic　responses　of　the　MCF-7　breast　cancer　cell　spheroid　(CCS)　to　DBP　exposure.　The　omics　results　showed　that　DBP　exposure　resulted　in　increased　glucose　and　glutamine　uptake　and　catabolism　in　breast　CCS　which　supported　the　production　of　nucleotides,　glycerophospholipids,　and　amino　acids,　providing　sufficient　energy　and　building　blocks　for　the　synthesis　of　DNA/RNA　and　the　cytomembrane　necessary　for　cell　proliferation.　These　results　were　further　corroborated　by　the　MSI　data,　showing　enhanced　abundances　of　ATP,　GMP,　UDP,　lysophosphatidylcholines,　phosphatidylcholines,　and　phosphatidylethanolamines　in　CCS　treated　with　DBP.　Interestingly,　most　of　these　biomolecules　were　predominantly　distributed　in　the　proliferative　region.　Our　result　indicated　that　the　enhanced　supply　of　energy　and　biosynthetic　substrates　in　the　peripheral　area　facilitated　the　proliferation　of　breast　CCS,　shedding　new　light　on　the　metabolic　mechanisms　of　DBP-promoted　breast　cancer　development.　©　2024　The　Authors.　Published　by　American　Chemical　Society