title: Proteomics study of open biopsy samples identifies peroxiredoxin 2 as a predictive biomarker of response to induction chemotherapy in osteosarcoma.
abstract: We attempted to identify biomarkers that would predict responsiveness of osteosarcoma (OS) to induction chemotherapy. Tumor tissues obtained by open biopsy before induction chemotherapy were investigated. On the basis of histological observations at the time of surgery and the Huvos grading system, 7 patients were classified as good responders and the other 6 as poor responders. Protein expression profiling was performed by two-dimensional difference gel electrophoresis. Among 3494 protein spots observed, the intensity of 33 spots was found to differ significantly between the two patient groups. The proteins for these 33 protein spots were identified by mass spectrometry. The higher expression of peroxiredoxin 2 (PRDX2) in poor responders was confirmed by Western blotting. Gene silencing assay demonstrated that reduced expression of PRDX2 was associated with increased sensitivity of OS cells to chemotherapeutic drugs such as methotrexate, doxorubicin and cisplatin. Moreover, siRNA-induced silencing of PRDX2 resulted in a decrease of cell proliferation, invasion and migration. These findings indicated that PRDX2 would be a candidate biomarker of response to induction chemotherapy. Measurement of PRDX2 in open biopsy samples before treatment may contribute to risk stratification therapy for OS.
  
  
  CONCLUSIONS
  The response of osteosarcoma patients to induction chemotherapy is critical because the prognosis of responders is quite favorable, whereas that of non-responders is poor. Although there are many therapeutic options for osteosarcoma, no parameter for predicting the response to induction chemotherapy has been available. We conducted a proteomics study aimed at developing a biomarker that would predict the response of osteosarcoma to induction chemotherapy. Using open biopsy samples obtained before chemotherapy, we conducted 2D-DIGE with our originally devised large-format electrophoresis apparatus and identified peroxiredoxin 2 (PRDX2) as a novel predictive biomarker. The diagnostic performance of PRDX2 was confirmed by ROC analysis, and its functional properties were investigated in a series of in vitro functional assays. Our findings indicate the possible application of PRDX2 as a predictive biomarker in patients with osteosarcoma.
authors.affiliation: Division of Pharmacoproteomics, National Cancer Center Research Institute, Tokyo, Japan; Department of Orthopedic Surgery, Juntendo University School of Medicine, Tokyo, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Kubota
authors.forename: Kenta
authors.initials: K
authors.lastname: Mukaihara
authors.forename: Akihiko
authors.initials: A
authors.lastname: Yoshida
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Tsuda
authors.forename: Akira
authors.initials: A
authors.lastname: Kawai
authors.forename: Tadashi
authors.initials: T
authors.lastname: Kondo
mesh_terms: D000293:Adolescent; D000328:Adult; D000970:Antineoplastic Agents / Q000494:pharmacology; D014408:Biomarkers, Tumor / Q000378:metabolism*; D001706:Biopsy; D001859:Bone Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D002648:Child; D019008:Drug Resistance, Neoplasm; D005260:Female; D020869:Gene Expression Profiling; D020868:Gene Silencing; D006801:Humans; D008297:Male; D009361:Neoplasm Invasiveness; D012516:Osteosarcoma / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism*; D054464:Peroxiredoxins / Q000378:metabolism*; D040901:Proteomics; D034741:RNA, Small Interfering / Q000378:metabolism; D055815:Young Adult
chemical_list: D000970:Antineoplastic Agents; D014408:Biomarkers, Tumor; D034741:RNA, Small Interfering; D054464:Peroxiredoxins
keywords: 2D-DIGE; Induction chemotherapy; Open biopsy; Osteosarcoma; Peroxiredoxin 2