title: Methotrexate induces DNA damage and inhibits homologous recombination repair in choriocarcinoma cells.
abstract: OBJECTIVE
  The objective of this study was to investigate the mechanism of sensitivity to methotrexate (MTX) in human choriocarcinoma cells regarding DNA damage response.
  
  
  METHODS
  Two choriocarcinoma cancer cell lines, JAR and JEG-3, were utilized in this study. An MTX-sensitive osteosarcoma cell line MG63, an MTX-resistant epithelial ovarian cancer cell line A2780 and an MTX-resistant cervical adenocarcinoma cell line Hela served as controls. Cell viability assay was carried out to assess MTX sensitivity of cell lines. MTX-induced DNA damage was evaluated by comet assay. Quantitative reverse transcription polymerase chain reaction was used to detect the mRNA levels of BRCA1, BRCA2, RAD51 and RAD52. The protein levels of γH2AX, RAD 51 and p53 were analyzed by Western blot.
  
  
  RESULTS
  Remarkable DNA strand breaks were observed in MTX-sensitive cell lines (JAR, JEG-3 and MG63) but not in MTX-resistant cancer cells (A2780 and Hela) after 48 h of MTX treatment. Only in the choriocarcinoma cells, the expression of homologous recombination (HR) repair gene RAD51 was dramatically suppressed by MTX in a dose- and time-dependent manner, accompanied with the increase in p53.
  
  
  CONCLUSIONS
  The MTX-induced DNA strand breaks accompanied by deficiencies in HR repair may contribute to the hypersensitivity to chemotherapy in choriocarcinoma.
authors.affiliation: Department of Obstetrics and Gynecology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology.
authors.forename: Lisha
authors.initials: L
authors.lastname: Xie
authors.affiliation: Department of Obstetrics and Gynecology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology; Department of Obstetrics and Gynecology, Central Hospital of Wuhan, Wuhan, China.
authors.forename: Tiancen
authors.initials: T
authors.lastname: Zhao
authors.affiliation: Department of Obstetrics and Gynecology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology.
authors.forename: Jing
authors.initials: J
authors.lastname: Cai
authors.affiliation: Department of Obstetrics and Gynecology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology.
authors.forename: You
authors.initials: Y
authors.lastname: Su
authors.affiliation: Department of Obstetrics and Gynecology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology.
authors.forename: Zehua
authors.initials: Z
authors.lastname: Wang
authors.affiliation: Department of Obstetrics and Gynecology, Union Hospital,Tongji Medical College, Huazhong University of Science and Technology.
authors.forename: Weihong
authors.initials: W
authors.lastname: Dong
keywords: DNA double-strand break; RAD51; chemotherapy hypersensitivity; choriocarcinoma; p53