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Tumor Biology

, Volume 36, Issue 5, pp 3735–3743 | Cite as

Construction of a plasmid for overexpression of human circadian gene period2 and its biological activity in osteosarcoma cells

  • An-yuan Cheng
  • Yan Zhang
  • Hong-jun Mei
  • Shuo Fang
  • Peng Ji
  • Jian Yang
  • Ling Yu
  • Wei-chun Guo
Research Article

Abstract

Beyond their established role in the mammalian circadian clock, recent studies have confirmed that the circadian genes have been implicated in tumor onset and progression. Currently, the biological effects of circadian genes on osteosarcoma cells’ proliferation and migration are not well understood. Period2 (Per2) is one of the core circadian genes that act as master regulators of development and is frequently dysregulated in several cancers. However, the effects of human Per2 (hPer2) on the biological behavior of osteosarcoma cells are rarely reported. In the present study, to address the expression of hPer2 in osteosarcoma cells, the pEGFP-N1-hPer2 eukaryotic expression vector was constructed and transfected into cultured MG63 cells using Lipofectamine™ 2000. The overexpression of hPer2 in MG63 cells was verified by qRT-PCR and Western blotting, respectively. Finally, we investigated the effects of hPer2 protein overexpression on MG63 cells’ viability, cycle, apoptosis, and invasive ability. In conclusion, the recombinant pEGFP-N1-hPer2 plasmid had been constructed successfully and expressed effectively in MG63 cells. Furthermore, results also showed that the viability, proliferation, and invasive abilities were suppressed, and the apoptosis was enhanced in MG63 cells. This preliminary study provides ground work for further research on the roles of circadian gene hPer2 in osteosarcoma cells MG63 and would offer promise for the development of novel therapeutic strategies in the treatment of osteosarcoma.

Keywords

Biological rhythm Circadian gene Per2 Osteosarcoma MG63 Proliferation Cell cycle Invasion 

Notes

Acknowledgments

We thank the Central Laboratory of the First Affiliated Hospital of Wuhan University. We also want to thank Ji-Shuang Zhu for his assistance in primer designing and Dr. Ling Yu for his careful review of the manuscript.

Conflicts of interest

None.

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • An-yuan Cheng
    • 1
  • Yan Zhang
    • 2
  • Hong-jun Mei
    • 3
  • Shuo Fang
    • 1
  • Peng Ji
    • 1
  • Jian Yang
    • 1
  • Ling Yu
    • 1
  • Wei-chun Guo
    • 1
  1. 1.Department of OrthopedicRenmin Hospital of Wuhan UniversityWuhanPeople’s Republic of China
  2. 2.Department of CardiologyThe General Hospital of Fengfeng Group. Co, LtdHandanPeople’s Republic of China
  3. 3.Department of OrthopedicWuhan No.5 HospitalWuhanPeople’s Republic of China

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