title: Proteomic analysis of the basic proteins in 5-fluorouracil resistance of human colon cancer cell line using the radical-free and highly reducing method of two-dimensional polyacrylamide gel electrophoresis.
abstract: 5-Fluorouracil (5-FU) is widely used for the treatment of patients with advanced colon cancers and it is the mainstay of chemotherapy. However, the acquisition of resistance to 5-FU is one of the most prominent obstacles to successful chemotherapy. The purpose of this study was to identify the novel biological basis of 5-FU resistance in colon cancer cells. This study is the first comparative proteomic analysis of basic proteins between the DLD-1 human colon cancer cell line and DLD-1/5-FU its 5-FU resistant sub-line using the radical-free and highly reducing method of two-dimensional polyacrylamide gel electrophoresis, which has a superior ability in the separation of basic proteins and the quantification of post-translational modification. A densitometric analysis was performed to quantify the modulated proteins, and protein spots showing significant changes were identified by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. Six basic proteins significantly modulated between DLD-1 and DLD-1/5-FU were identified. All of them showed up-regulated expression in DLD-1/5-FU in comparison to DLD-1. The six identified spots, corresponding to five different proteins included heterogeneous nuclear ribonucleoprotein G, mitochondrial transcription factor A, histone H2B, histone H4 and ribosomal protein L3. Among the 5 basic proteins, several proteins are potentially related to 5-FU resistance by protecting the cells from DNA damage.
authors.affiliation: Department of General and Gastroenterological Surgery, Osaka Medical College, Osaka, Takatsuki City, Osaka 569-8686, Japan.
authors.forename: Satoru
authors.initials: S
authors.lastname: Tanaka
authors.forename: Akiko
authors.initials: A
authors.lastname: Sakai
authors.forename: Kousei
authors.initials: K
authors.lastname: Kimura
authors.forename: Hideji
authors.initials: H
authors.lastname: Yoshida
authors.forename: Hideo
authors.initials: H
authors.lastname: Fushitani
authors.forename: Akihito
authors.initials: A
authors.lastname: Ogata
authors.forename: Akiko
authors.initials: A
authors.lastname: Miyamoto
authors.forename: Masakazu
authors.initials: M
authors.lastname: Fukushima
authors.forename: Akira
authors.initials: A
authors.lastname: Wada
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Tanigawa
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000378:metabolism*; D019008:Drug Resistance, Neoplasm / Q000502:physiology*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005472:Fluorouracil / Q000494:pharmacology*; D006801:Humans; D040901:Proteomics / Q000379:methods*; D000090822:Ribosomal Protein L3; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D000970:Antineoplastic Agents; C000716547:RPL3 protein, human; D000090822:Ribosomal Protein L3; D005472:Fluorouracil