title: In situ Proteomic Profiling of Curcumin Targets in HCT116 Colon Cancer Cell Line.
abstract: To date, the exact targets and mechanism of action of curcumin, a natural product with anti-inflammatory and anti-cancer properties, remain elusive. Here we synthesized a cell permeable curcumin probe (Cur-P) with an alkyne moiety, which can be tagged with biotin for affinity enrichment, or with a fluorescent dye for visualization of the direct-binding protein targets of curcumin in situ. iTRAQ(TM) quantitative proteomics approach was applied to distinguish the specific binding targets from the non-specific ones. In total, 197 proteins were confidently identified as curcumin binding targets from HCT116 colon cancer cell line. Gene Ontology analysis showed that the targets are broadly distributed and enriched in the nucleus, mitochondria and plasma membrane, and they are involved in various biological functions including metabolic process, regulation, response to stimulus and cellular process. Ingenuity Pathway Analysis(TM) (IPA) suggested that curcumin may exert its anticancer effects over multiple critical biological pathways including the EIF2, eIF4/p70S6K, mTOR signaling and mitochondrial dysfunction pathways. Functional validations confirmed that curcumin downregulates cellular protein synthesis, and induces autophagy, lysosomal activation and increased ROS production, thus leading to cell death.
authors.affiliation: Interdisciplinary Research Group in Infectious Diseases, Singapore-MIT Alliance for Research &Technology (SMART), 138602, Singapore.|Department of Biological Sciences, National University of Singapore, 117543, Singapore.|The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Sciences, Nanjing University, Nanjing, 210023, China.
authors.forename: Jigang
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 117597, Singapore.
authors.forename: Jianbin
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Department of Chemical and Biomolecular Engineering, National University of Singapore, 117585, Singapore.
authors.forename: Chong-Jing
authors.initials: CJ
authors.lastname: Zhang
authors.affiliation: Department of Biological Sciences, National University of Singapore, 117543, Singapore.
authors.forename: Yin Kwan
authors.initials: YK
authors.lastname: Wong
authors.affiliation: Department of Biological Sciences, National University of Singapore, 117543, Singapore.
authors.forename: Teck Kwang
authors.initials: TK
authors.lastname: Lim
authors.affiliation: The State Key Laboratory of Pharmaceutical Biotechnology, College of Life Sciences, Nanjing University, Nanjing, 210023, China.
authors.forename: Zi-Chun
authors.initials: ZC
authors.lastname: Hua
authors.affiliation: Department of Chemical and Biomolecular Engineering, National University of Singapore, 117585, Singapore.
authors.forename: Bin
authors.initials: B
authors.lastname: Liu
authors.affiliation: Interdisciplinary Research Group in Infectious Diseases, Singapore-MIT Alliance for Research &Technology (SMART), 138602, Singapore.|Departments of Biological Engineering &Chemistry, Massachusetts Institute of Technology, 02139, United States of America.
authors.forename: Steven R
authors.initials: SR
authors.lastname: Tannenbaum
authors.affiliation: Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, 117597, Singapore.
authors.forename: Han-Ming
authors.initials: HM
authors.lastname: Shen
authors.affiliation: Department of Biological Sciences, National University of Singapore, 117543, Singapore.
authors.forename: Qingsong
authors.initials: Q
authors.lastname: Lin
mesh_terms: D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001343:Autophagy / Q000187:drug effects*; D001665:Binding Sites; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003110:Colonic Neoplasms / Q000188:drug therapy*; D003474:Curcumin / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000378:metabolism; D045325:HCT116 Cells; D006801:Humans; D008247:Lysosomes / Q000187:drug effects; D008928:Mitochondria / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects*; D040901:Proteomics / Q000379:methods; D017382:Reactive Oxygen Species / Q000378:metabolism; D038762:Ribosomal Protein S6 Kinases, 70-kDa / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D000970:Antineoplastic Agents; D004268:DNA-Binding Proteins; C111931:ELF4 protein, human; D017382:Reactive Oxygen Species; D014157:Transcription Factors; C546842:MTOR protein, human; D038762:Ribosomal Protein S6 Kinases, 70-kDa; D058570:TOR Serine-Threonine Kinases; D003474:Curcumin