title: Protein expression changes induced in a malignant melanoma cell line by the curcumin analogue compound D6.
abstract: We have previously demonstrated that the hydroxylated biphenyl compound D6 (3E,3'E)-4,4'-(5,5',6,6'-tetramethoxy-[1,1'-biphenyl]-3,3'-diyl)bis(but-3-en-2-one), a structural analogue of curcumin, exerts a strong antitumor activity on melanoma cells both in vitro and in vivo. Although the mechanism of action of D6 is yet to be clarified, this compound is thought to inhibit cancer cell growth by arresting the cell cycle in G2/M phase, and to induce apoptosis through the mitochondrial intrinsic pathway. To investigate the changes in protein expression induced by exposure of melanoma cells to D6, a differential proteomic study was carried out on D6-treated and untreated primary melanoma LB24Dagi cells.
  
  
  
  Proteins were fractionated by SDS-PAGE and subjected to in gel digestion. The peptide mixtures were analyzed by liquid chromatography coupled with tandem mass spectrometry. Proteins were identified and quantified using database search and spectral counting. Proteomic data were finally uploaded into the Ingenuity Pathway Analysis software to find significantly modulated networks and pathways.
  
  
  
  Analysis of the differentially expressed protein profiles revealed the activation of a strong cellular stress response, with overexpression of several HSPs and stimulation of ubiquitin-proteasome pathways. These were accompanied by a decrease of protein synthesis, evidenced by downregulation of proteins involved in mRNA processing and translation. These findings are consistent with our previous results on gene expression profiling in melanoma cells treated with D6.
  
  
  
  Our findings confirm that the curcumin analogue D6 triggers a strong stress response in melanoma cells, turning down majority of cell functions and finally driving cells to apoptosis.
authors.affiliation: Institute of Biomolecular Chemistry, National Research Council of Italy, Traversa la Crucca, 3, 07100, Sassari, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Pisano
authors.affiliation: Proteomics Laboratory, Porto Conte Ricerche, Tramariglio, Alghero, Italy.|Biosistema Scrl, Sassari, Italy.
authors.forename: Antonio
authors.initials: A
authors.lastname: Palomba
authors.affiliation: Proteomics Laboratory, Porto Conte Ricerche, Tramariglio, Alghero, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Tanca
authors.affiliation: Proteomics Laboratory, Porto Conte Ricerche, Tramariglio, Alghero, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Pagnozzi
authors.affiliation: Proteomics Laboratory, Porto Conte Ricerche, Tramariglio, Alghero, Italy.
authors.forename: Sergio
authors.initials: S
authors.lastname: Uzzau
authors.affiliation: Proteomics Laboratory, Porto Conte Ricerche, Tramariglio, Alghero, Italy.
authors.forename: Maria Filippa
authors.initials: MF
authors.lastname: Addis
authors.affiliation: Institute of Biomolecular Chemistry, National Research Council of Italy, Traversa la Crucca, 3, 07100, Sassari, Italy.
authors.forename: Maria Antonietta
authors.initials: MA
authors.lastname: Dettori
authors.affiliation: Institute of Biomolecular Chemistry, National Research Council of Italy, Traversa la Crucca, 3, 07100, Sassari, Italy.
authors.forename: Davide
authors.initials: D
authors.lastname: Fabbri
authors.affiliation: Institute of Biomolecular Chemistry, National Research Council of Italy, Traversa la Crucca, 3, 07100, Sassari, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Palmieri
authors.affiliation: Institute of Biomolecular Chemistry, National Research Council of Italy, Traversa la Crucca, 3, 07100, Sassari, Italy. carla.rozzo@icb.cnr.it.
authors.forename: Carla
authors.initials: C
authors.lastname: Rozzo
mesh_terms: D001713:Biphenyl Compounds / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003474:Curcumin / Q000031:analogs & derivatives* / Q000494:pharmacology; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D053263:Gene Regulatory Networks / Q000187:drug effects*; D006801:Humans; D008545:Melanoma / Q000188:drug therapy / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015398:Signal Transduction / Q000187:drug effects
chemical_list: D001713:Biphenyl Compounds; D003474:Curcumin
keywords: Curcumin; Hydroxylated biphenyls; Melanoma cells; Molecular pathways analysis; Proteomic profiling