Chinese Journal of Tissue Engineering Research ›› 2015, Vol. 19 ›› Issue (40): 6408-6412.doi: 10.3969/j.issn.2095-4344.2015.40.003

Previous Articles     Next Articles

Establishment of osteosarcoma cell models: effects of NS-398 on invasive ability of MG-63 cells under hypoxic condition 

Wang Yi-fan1, Xu Yong-tao2   

  1. 1Medical School of Yangtze University, Jingzhou 434020, Hubei Province, China; 2Third Department of Orthopedics, Jingzhou Central Hospital, Jingzhou 434020, Hubei Province, China
  • Online:2015-09-30 Published:2015-09-30
  • Contact: Xu Yong-tao, Third Department of Orthopedics, Jingzhou Central Hospital, Jingzhou 434020, Hubei Province, China
  • About author:Wang Yi-fan, Medical School of Yangtze University, Jingzhou 434020, Hubei Province, China

PDF

342

Abstract:

BACKGROUND: In anoxic environment, invasive ability of tumor cells significantly increased. The increase in above invasive ability is possibly associated with the changes in urokinase or matrix metalloproteinases. Studies on hypoxia and tumor therapy especially the effect on chemotherapy have made great progress, and found that hypoxia-inducible factor 1 is a key nuclear factor mediating cellular hypoxia response, and also an important factor for regulating cell apoptosis during hypoxia.

OBJECTIVE: To observe the effects of cyclooxygenase-2 inhibitor NS-398 on invasive ability of MG-63 cells under hypoxic condition in cell models of hypoxia osteosarcoma.

METHODS: Chemical hypoxia inducer cobalt chloride was used to establish chemical hypoxia environment of concentration gradient of 0, 100, 200, 400 μmol/L. Under 200 μmol/L cobalt chloride, concentration gradient of 30, 60, 90 μmol/L of NS-398 were set. Osteosarcoma cells were determined after 48 hours of culture. Quantitative PCR was used to detect the expression of matrix metalloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA. Western blot assay was applied to measure hypoxia-inducible factor 1α expression. Flow cytometry was utilized to examine cell apoptosis in each group.

RESULTS AND CONCLUSION: PCR and western blot assay results demonstrated that with increased concentration of cobalt chloride, matrix metalloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA and protein expression increased (P < 0.05), showing a concentration-dependent manner. Under cobalt chloride concentration of 200 μmol/L and hypoxia, with increased concentration of NS-398, matrix metalloproteinases 9 mRNA and hypoxia-inducible factor 1α mRNA and protein expression gradually reduced, showing a concentration-dependent manner (P < 0.05). Flow cytometry results showed that under 400 μmol/L cobalt chloride concentration, cell apoptosis noticeably reduced. Under cobalt chloride concentration of 200 μmol/L and hypoxia, with increased concentration of NS-398, cell apoptosis increased (P < 0.05). Results confirm that anaerobic environment is an important factor for increased invasive ability of osteosarcoma. Under the anaerobic environment, NS-398 could effectively suppress the invasive ability of osteosarcoma and promote cell apoptosis.

Key words: Tissue Engineering, Anoxia, Osteosarcoma, Apoptosis

CLC Number: