Lung　adenocarcinoma,　the　most　common　type　of　lung　cancer,　has　increased　in　recent　years.　Prognosis　is　still　poor,　and　pathogenesis　remains　unclear.　This　study　aimed　to　investigate　the　membrane　protein　profile　differences　between　lung　adenocarcinoma　and　normal　tissue.　Manual　microdissection　was　used　to　isolate　the　target　cells　from　tumor　tissue　and　normal　tissue.　iTRAQ　labeling　combined　with　2D-LC-MS/MS　yielded　a　differential　expression　profile　of　membrane　proteins.　Bioinformatic　analysis　was　performed　using　Gene　Ontology,　WEGO,　PID,　and　KEGG.　S100A14　protein　was　selectively　verified　by　Western　blotting.　The　relationship　of　S100A14　expression　with　clinicopathological　features　in　lung　cancer　patients　was　evaluated　using　immunohistochemistry.　As　a　result,　568　differential　proteins　were　identified;　257　proteins　were　upregulated　and　311　were　downregulated.　Of　these　proteins,　48%　were　found　to　be　membrane　bound　or　membrane　associated.　These　proteins　enable　the　physiological　functions　of　binding,　catalysis,　molecular　transduction,　transport,　and　molecular　structure.　For　these　differential　proteins,　35　pathways　were　significantly　enriched　through　the　Pathway　Interaction　Database,　whereas　19　pathways　were　enriched　via　KEGG.　The　overexpression　and　cellular　distribution　of　S100A14　in　lung　cancer　were　confirmed.　We　found　that　upregulation　of　S100A14　was　associated　with　well　or　moderate　differentiation.　The　iTRAQ-coupled　2D-LC-MS/MS　technique　is　a　potential　method　for　comparing　membrane　protein　profiles　between　tumor　and　normal　tissue.　Such　analysis　may　also　help　in　identifying　novel　biomarkers　and　the　mechanisms　underlying　carcinogenesis.