title: Hypoxia promotes drug resistance in osteosarcoma cells via activating AMP-activated protein kinase (AMPK) signaling.
abstract: OBJECTIVE
  Drug resistance has been recognized to be a major obstacle to the chemotherapy for osteosarcoma. And the potential importance of hypoxia as a target to reverse drug resistance in osteosarcoma has been indicated, though the mechanism underlining such role is not clarified. The present study aims to investigate the role of hypoxia in the drug resistance in osteosarcoma cells via activating AMP-activated protein kinase (AMPK) signaling.
  
  
  METHODS
  We investigated the promotion of the resistance to doxorubicin of osteosarcoma MG-63 and U2-os cells in vitro, and then determined the role of hypoxia-inducible factor-1 (HIF-1)α and HIF-1β, the activation and regulatory role of AMPK in the osteosarcoma U2-os cells which were treated with doxorubicin under hypoxia.
  
  
  RESULTS
  It was demonstrated that hypoxia significantly reduced the sensitivity of MG-63 and U2-os cells to doxorubicin, indicating an inhibited viability reduction and a reduced apoptosis promotion. And such reduced sensitivity was not associated with HIF-1α, though it was promoted by hypoxia in U2-os cells. Interestingly, the AMPK signaling was significantly promoted by hypoxia in the doxorubicin-treated U2-os cells, with a marked upregulation of phosphorylated AMPK (Thr 172) and phosphorylated acetyl-CoA carboxylase (ACC) (Ser 79), which were sensitive to the AMPK activator, AICAR and the AMPK inhibitor, Compound C. Moreover, the promoted AMPK activity by AICAR or the downregulated AMPK activity by Compound C significantly reduced or promoted the sensitivity of U2-os cells to doxorubicin.
  
  
  CONCLUSIONS
  The present study confirmed the AMPK signaling activation in the doxorubicin-treated osteosarcoma cells, in response to hypoxia, and the chemical upregulation or downregulation of AMPK signaling reduced or increased the chemo-sensitivity of osteosarcoma U2-os cells in vitro. Our study implies that AMPK inhibition might be a effective strategy to sensitize osteocarcoma cells to chemotherapy.
authors.affiliation: Department of Orthopedics, China-Japan Union Hospital of Jilin University, No. 126 of Xiantai Street, Erdao District, Changchun 130033, China.
authors.forename: Changfu
authors.initials: C
authors.lastname: Zhao
authors.affiliation: Department of Orthopedics, China-Japan Union Hospital of Jilin University, No. 126 of Xiantai Street, Erdao District, Changchun 130033, China.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Department of Orthopedics, Second Hospital of Jilin University, Changchun 130000, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Yu
authors.affiliation: Department of Orthopedics, Second Hospital of Jilin University, Changchun 130000, China.
authors.forename: Shudong
authors.initials: S
authors.lastname: Sun
authors.affiliation: Department of Orthopedics, China-Japan Union Hospital of Jilin University, No. 126 of Xiantai Street, Erdao District, Changchun 130033, China.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, China-Japan Union Hospital of Jilin University, No. 126 of Xiantai Street, Erdao District, Changchun 130033, China.
authors.forename: Guangyao
authors.initials: G
authors.lastname: Liu
keywords: AMPK signaling; Doxorubicin; Hypoxia; Osteosarcoma cells; Resistance