title: Comparative proteomic analysis of the ribosomes in 5-fluorouracil resistance of a human colon cancer cell line using the radical-free and highly reducing method of two-dimensional polyacrylamide gel electrophoresis.
abstract: Many auxiliary functions of ribosomal proteins (r-proteins) have received considerable attention in recent years. However, human r-proteins have hardly been examined by proteomic analysis. In this study, we isolated ribosomal particles and subsequently compared the proteome of r-proteins between the DLD-1 human colon cancer cell line and its 5-fluorouracil (5-FU)-resistant sub-line, DLD-1/5-FU, using the radical-free and highly reducing method of two-dimensional polyacrylamide gel electrophoresis, which has a superior ability to separate basic proteins, and we discuss the role of r-proteins in 5-FU resistance. Densitometric analysis was performed to quantify modulated proteins, and protein spots showing significant changes were identified by employing matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Three basic proteins (L15, L37 and prohibitin) which were significantly modulated between DLD-1 and DLD-1/5-FU were identified. Two proteins, L15 and L37, showed down-regulated expression in DLD-1/5-FU in comparison to DLD-1. Prohibitin, which is not an r-protein and is known to be localized in the mitochondria, showed up-regulated expression in DLD-1/5-FU. These 3 proteins may be related to 5-FU resistance.
authors.affiliation: Department of General and Gastroenterological Surgery, Osaka Medical College, Takatsuki City, Osaka 569-8686, Japan.
authors.forename: Kosei
authors.initials: K
authors.lastname: Kimura
authors.forename: Akira
authors.initials: A
authors.lastname: Wada
authors.forename: Masami
authors.initials: M
authors.lastname: Ueta
authors.forename: Akihiko
authors.initials: A
authors.lastname: Ogata
authors.forename: Satoru
authors.initials: S
authors.lastname: Tanaka
authors.forename: Akiko
authors.initials: A
authors.lastname: Sakai
authors.forename: Hideji
authors.initials: H
authors.lastname: Yoshida
authors.forename: Hideo
authors.initials: H
authors.lastname: Fushitani
authors.forename: Akiko
authors.initials: A
authors.lastname: Miyamoto
authors.forename: Masakazu
authors.initials: M
authors.lastname: Fukushima
authors.forename: Toshio
authors.initials: T
authors.lastname: Uchiumi
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Tanigawa
mesh_terms: D017209:Apoptosis / Q000502:physiology; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D040901:Proteomics / Q000379:methods*; D012269:Ribosomal Proteins / Q000032:analysis*; D012270:Ribosomes / Q000737:chemistry
chemical_list: D012269:Ribosomal Proteins