title: XPA versus ERCC1 as chemosensitising agents to cisplatin and mitomycin C in prostate cancer cells: role of ERCC1 in homologous recombination repair.
abstract: Nucleotide excision repair is the principal mechanism for the removal of bulky DNA adducts caused by a range of chemotherapeutic drugs, and contributes to cisplatin resistance. In this study, we used synthetic siRNAs targeted to XPA and ERCC1 and compared their effectiveness in sensitising mismatch repair deficient prostate cancer cell lines to cisplatin and mitomycin C. Downregulation of ERCC1 sensitised DU145 and PC3 cells to cisplatin and mitomycin C. In contrast, XPA downregulation did not sensitise either cell line to mitomycin C, and only sensitised DU145 cells to cisplatin. The effects of ERCC1 downregulation may be due to its role in homologous recombination repair. Excision repair of cisplatin adducts in PC3 cells was attenuated to a similar extent by XPA and ERCC1 downregulation. Downregulation of XPA but not ERCC1 caused an increase in the number of cisplatin-induced RAD51 foci in PC3 cells, suggesting that HRR is able to substitute for NER in these cells. We observed co-localisation of ERCC1 and RAD51 in cisplatin treated PC3 cells by immunofluorescence and co-immunoprecipitation, which may represent recruitment of ERCC1/XPF to sites of recombination repair. These results indicate that ERCC1 is a broader therapeutic target than XPA with which to sensitise cancer cells to chemotherapy because of its additional role in recombination repair.
authors.affiliation: Prostate Cancer Research Centre, Institute of Urology, University College London, and St. Bartholomew's Hospital, Department of Medical Oncology, London W1W 7EJ, UK.
authors.forename: Michele
authors.initials: M
authors.lastname: Cummings
authors.forename: Karen
authors.initials: K
authors.lastname: Higginbottom
authors.forename: Claire J
authors.initials: CJ
authors.lastname: McGurk
authors.forename: Oscar Gee-Wang
authors.initials: OG
authors.lastname: Wong
authors.forename: Beate
authors.initials: B
authors.lastname: Köberle
authors.forename: R Timothy D
authors.initials: RT
authors.lastname: Oliver
authors.forename: John R
authors.initials: JR
authors.lastname: Masters
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000235:genetics*; D004720:Endonucleases / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D006801:Humans; D047468:Immunoprecipitation; D008297:Male; D016685:Mitomycin / Q000494:pharmacology*; D011471:Prostatic Neoplasms / Q000473:pathology*; D034741:RNA, Small Interfering; D011995:Recombination, Genetic; D051760:Xeroderma Pigmentosum Group A Protein / Q000235:genetics*
chemical_list: D000970:Antineoplastic Agents; D004268:DNA-Binding Proteins; D034741:RNA, Small Interfering; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein; D016685:Mitomycin; C071447:ERCC1 protein, human; D004720:Endonucleases; D002945:Cisplatin