Tumor Biology

, Volume 37, Issue 2, pp 1591–1598 | Cite as

Oxidative stress induced by low-dose doxorubicin promotes the invasiveness of osteosarcoma cell line U2OS in vitro

  • Seung Han Shin
  • Young Joon Choi
  • Hyewon Lee
  • Han-Soo Kim
  • Sung Wook Seo
Research Article


Reactive oxygen species (ROS) are known to mediate doxorubicin (DOX)-induced apoptosis and are the major cause of DOX toxicity. We introduce a novel in vitro phenomenon of osteosarcoma (OS) cell line caused by low-dose DOX-induced oxidative stress. Human osteosarcoma cell line U2OS was used for the experiments. Hydrogen peroxide (H2O2) and the antioxidant compound N-acetylcysteine (NAC) were used to investigate the involvement of oxidative stress. In proliferation assays, low dose of DOX (below 200 nM) did not affect U2OS proliferation significantly for up to 48 h. In MatrigelTM invasion assay, DOX increased the invasiveness of U2OS at around 100 nM, which is a subclinical concentration. Quantitative real-time polymerase chain reaction and gelatin zymography showed increased MMP-9 expression and increased MMP-9 enzymatic activity, respectively, in the presence of DOX doses that increased the invasiveness of U2OS. H2O2, a representative source of ROS, also increased the invasiveness of U2OS as DOX did, with similar patterns. However, when the cells were pre-treated with NAC, no DOX- or H2O2-mediated increase of invasiveness or MMP-9 expression was evident. The results suggest that oxidative stress induced by low-dose DOX promotes the invasiveness of osteosarcoma cell line U2OS in vitro, through MMP-9 induction.


Oxidative stress Reactive oxygen species Doxorubicin Osteosarcoma Invasiveness MMP-9 



This study was supported by a grant of the Korean Health Technology R&D Project, Ministry of Health & Welfare, Republic of Korea (HI12C1065).

Conflicts of interest



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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  1. 1.Department of Orthopedic Surgery, Seoul St. Mary’s Hospital, College of MedicineThe Catholic University of KoreaSeoulSouth Korea
  2. 2.Department of Health Sciences and Technology, SAIHSTSungkyunkwan UniversitySeoulSouth Korea
  3. 3.Department of Orthopedic SurgerySeoul National University Hospital, Seoul National University College of MedicineSeoulSouth Korea
  4. 4.Department of Orthopedic Surgery, Samsung Medical CenterSungkyunkwan University School of MedicineSeoulSouth Korea

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