title: Establishment and characterization of two 5-fluorouracil-resistant hepatocellular             carcinoma cell lines.
abstract: 5-Fluorouracil (5-FU) chemotherapy is the first choice treatment for advanced             hepatocellular carcinoma (HCC), and resistance is the major obstacle to successful             treatment. Recent studies have reported that epithelial-to-mesenchymal transition             (EMT) is associated with chemoresistance in cancers. We speculated that EMT and             5-FU metabolism are related to the mechanism of 5-FU resistance. First, two 5-FU-resistant             cell lines, HLF-R4 and HLF-R10, were established from the HLF undifferentiated             human HCC cell line. Whereas cell growth was similar in the HLF and HLF-R cell             lines, HLF-Rs are about 4- and 10-fold more resistant compared with the HLF cells;             thus, we named these cell lines HLF-R4 and HLF-R10, respectively. The terminal             deoxyribonucleotidyl transferase-mediated dUTP nick end labeling assay also showed             a dramatically decreased number of apoptotic cells in the HLF-Rs after treatment             with 5-FU. We next assessed the characteristics of the HLF, HLF-R4 and HLF-R10             cells. Consistent with our hypothesis, the HLF-Rs had typical morphologic phenotypes             of EMT, loss of cell-cell adhesion, spindle-shaped morphology and increased formation             of pseudopodia. Real-time quantitative reverse transcriptase polymerase chain             reaction data showed downregulated E-cadherin and upregulated Twist-1 and also             indicated that EMT changes occurred in the HLF-Rs. We also found decreased ribonucleotide             reductase and increased multidrug resistance protein 5 genes in the HLF-R cells.             Our results suggested that the metabolism of EMT and 5-FU has important roles             in 5-FU chemoresistance in the HLF-R cells, and that the HLF-R cells would be             useful in vitro models for understanding the 5-FU-resistant mechanisms in HCC.
authors.affiliation: Department of Clinical Education and Research, Graduate School                     of Pharmaceutical Sciences, Chiba University, Chiba 260-8670, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Uchibori
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kasamatsu
authors.forename: Masahiko
authors.initials: M
authors.lastname: Sunaga
authors.forename: Satoshi
authors.initials: S
authors.lastname: Yokota
authors.forename: Tomoya
authors.initials: T
authors.lastname: Sakurada
authors.forename: Eriko
authors.initials: E
authors.lastname: Kobayashi
authors.forename: Masaharu
authors.initials: M
authors.lastname: Yoshikawa
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Uzawa
authors.forename: Shiro
authors.initials: S
authors.lastname: Ueda
authors.forename: Hideki
authors.initials: H
authors.lastname: Tanzawa
authors.forename: Nobunori
authors.initials: N
authors.lastname: Sato
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D048708:Cell Growth Processes / Q000187:drug effects; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm; D058750:Epithelial-Mesenchymal Transition; D005472:Fluorouracil / Q000494:pharmacology*; D006801:Humans; D020287:In Situ Nick-End Labeling / Q000379:methods; D008113:Liver Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology
chemical_list: D000964:Antimetabolites, Antineoplastic; D005472:Fluorouracil