title: Oxaliplatin induces different cellular and molecular chemoresistance patterns in colorectal cancer cell lines of identical origins.
abstract: BACKGROUND
  Cancer cells frequently adopt cellular and molecular alterations and acquire resistance to cytostatic drugs. Chemotherapy with oxaliplatin is among the leading treatments for colorectal cancer with a response rate of 50%, inducing intrastrand cross-links on the DNA. Despite of this drug's efficiency, resistance develops in nearly all metastatic patients. Chemoresistance being of crucial importance for the drug's clinical efficiency this study aimed to contribute to the identification and description of some cellular and molecular alterations induced by prolonged oxaliplatin therapy. Resistance to oxaliplatin was induced in Colo320 (Colo320R) and HT-29 (HT-29R) colorectal adenocarcinoma cell lines by exposing the cells to increasing concentrations of the drug. Alterations in morphology, cytotoxicity, DNA cross-links formation and gene expression profiles were assessed in the parental and resistant variants with microscopy, MTT, alkaline comet and pangenomic microarray assays, respectively.
  
  
  RESULTS
  Morphology analysis revealed epithelial-to-mesenchymal transition in the resistant vs parental cells suggesting alterations of the cells' adhesion complexes, through which they acquire increased invasiveness and adherence. Cytotoxicity measurements demonstrated resistance to oxaliplatin in both cell lines; Colo320 being more sensitive than HT-29 to this drug (P < 0.001). The treatment with oxaliplatin caused major DNA cross-links in both parental cell lines; in Colo320R small amounts of DNA cross-links were still detectable, while in HT-29R not. We identified 441 differentially expressed genes in Colo320R and 613 in HT-29R as compared to their parental counterparts (at least 1.5 -fold up- or down- regulation, p < 0.05). More disrupted functions and pathways were detected in HT-29R cell line than in Colo320R, involving genes responsible for apoptosis inhibition, cellular proliferation and epithelial-to-mesenchymal transition. Several upstream regulators were detected as activated in HT-29R cell line, but not in Colo320R.
  
  
  CONCLUSIONS
  Our findings revealed a more resistant phenotype in HT-29R as compared to Colo320R and different cellular and molecular chemoresistance patterns induced by prolonged treatment with oxaliplatin in cell lines with identical origins (colorectal adenocarcinomas).
authors.affiliation: The Oncology Institute Prof.Dr.I. Chiricuta, 400015 Republicii Str,, nr, 34-36, Cluj-Napoca, Romania. vpiroska@yahoo.com
authors.forename: Piroska
authors.initials: P
authors.lastname: Virag
authors.forename: Eva
authors.initials: E
authors.lastname: Fischer-Fodor
authors.forename: Maria
authors.initials: M
authors.lastname: Perde-Schrepler
authors.forename: Ioana
authors.initials: I
authors.lastname: Brie
authors.forename: Corina
authors.initials: C
authors.lastname: Tatomir
authors.forename: Loredana
authors.initials: L
authors.lastname: Balacescu
authors.forename: Ioana
authors.initials: I
authors.lastname: Berindan-Neagoe
authors.forename: Bogdan
authors.initials: B
authors.lastname: Victor
authors.forename: Ovidiu
authors.initials: O
authors.lastname: Balacescu
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects / Q000235:genetics; D015179:Colorectal Neoplasms / Q000473:pathology*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D006801:Humans; D009361:Neoplasm Invasiveness; D009944:Organoplatinum Compounds / Q000494:pharmacology*; D000077150:Oxaliplatin; D010641:Phenotype; D015203:Reproducibility of Results; D013997:Time Factors; D059467:Transcriptome / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D003432:Cross-Linking Reagents; D009944:Organoplatinum Compounds; D000077150:Oxaliplatin