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Tumor Biology

, Volume 37, Issue 7, pp 8869–8877 | Cite as

The tumor suppressive role of RASSF1A in osteosarcoma through the Wnt signaling pathway

  • Wei-Guo Wang
  • Shi-Jie Chen
  • Jin-Shen He
  • Jing-Song Li
  • Xiao-Fang Zang
Original Article

Abstract

Ras-association domain family 1 isoform A (RASSF1A) is a tumor suppressor gene and its expression is lost in numerous types of cancer cells, including primary osteosarcoma cells. However, its functional significance in osteosarcoma has not been well defined. The messenger RNA (mRNA) expression of RASSF1A in osteosarcoma tissues and corresponding non-tumoral tissues was measured by real-time PCR. Overexpression of RASSF1A was established by an adenoviral vector expressing RASSF1A. Cell migration and invasion were analyzed in transwells. Apoptosis and cell cycle were analyzed using flow cytometry. Wnt/β-catenin activity was measured by TCF reporter dual-luciferase assay. Cell viability was measured by MTT assay. Protein expression was detected by Western blot. RASSF1A mRNA expression was significantly lower in osteosarcoma tissues than that in the corresponding non-tumoral tissues. The lowered RASSF1A expression correlated with the clinical severity of osteosarcoma. rAd-RASSF1A injection significantly inhibited the growth of xenograft MNNG/HOS tumors in mice. Overexpression of RASSF1A resulted in significant inhibition of the proliferation, migration, and invasion; induced apoptosis; and arrested cell cycle at G0/G1 phase in both the MNNG/HOS and SaOS2 cells. Overexpression of RASSF1A inhibited the Wnt/β-catenin activity, decreased phosphorylation of Akt/glycogen synthase kinase-3-β (GSK3-β), and increased phosphorylation of mammalian sterile 20-like kinase 1 (MST1). Overexpression of RASSF1A downregulated the cyclin D1, c-Myc, and matrix metalloproteinase-7 (MMP-7) protein levels. RASSF1A functions as a tumor suppressor in osteosarcoma and exerts anti-cancer roles through regulating Akt/GSK-3-Wnt/β-catenin signaling.

Keywords

Osteosarcoma Wnt RASSF1A β-catenin 

Notes

Compliance with ethical standard

The study was pre-approved by the Institutional Ethics Committee of the Third Xiangya Hospital and written informed consent was obtained from all patients.

Conflicts of interest

None

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Wei-Guo Wang
    • 1
  • Shi-Jie Chen
    • 1
  • Jin-Shen He
    • 1
  • Jing-Song Li
    • 1
  • Xiao-Fang Zang
    • 1
  1. 1.Department of Orthopaedic SurgeryThe Third Xiangya Hospital of Central South UniversityChangshaPeople’s Republic of China

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