Chinese Journal of Tissue Engineering Research ›› 2011, Vol. 15 ›› Issue (36): 6657-6660.doi: 10.3969/j.issn.1673-8225.2011.36.002

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Effect of DNA demethylation on telomerase reverse transcriptase expression in rat bone marrow mesenchymal stem cells

Zhao Hong-xian1, Lu Wei1, Guo Yong1, Chen Xia2   

  1. 1Department of Histology and Embryology, Luzhou Medical College, Luzhou  646000, Sichuan Province, China; 2Department of Gastroenterology, Affiliated Hospital of Luzhou Medical College, Luzhou  646000, Sichuan Province, China
  • Received:2011-03-01 Revised:2011-04-15 Online:2011-09-03 Published:2011-09-03
  • Contact: Chen Xia, Master, Department of Gastroenterology, Affiliated Hospital of Luzhou Medical College, Luzhou 646000, Sichuan Province, China flygirl0012000@yahoo.com.cn
  • About author:Zhao Hong-xian★, Master, Lecturer, Department of Histology and Embryology, Luzhou Medical College, Luzhou 646000, Sichuan Province, China zilong5276@sohu.com

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Abstract:

BACKGROUND: DNA demethylation is an important epigenetic modification, which plays a vital role in telomerase regualtion of tumor cells. It is not clear to effect of DNA demethylation on telomerase activity in bone marrow mesenchymal stem cells.
OBJECTIVE: To observe the effect of DNA demethylation on proliferation and telomerase reverse transcriptase expression in bone marrow mesenchymal stem cells.
METHODS: Bone marrow mesenchymal stem cells were isolated by using the method of adhesive culture of whole bone marrow. In the third passage bone marrow mesenchymal stem cells, 5-azacytidine was added to 0, 3, 6, 12, 24 μmol/L. CD44, CD45 and telomerase reverse transcriptase were dectcted by immunocytochemistry, and effect of DNA demethylation on proliferation was detected by MTT method at 1, 2, 3, 5, 7 days after 5-azacytidine intervention.
RESULTS AND CONCLUSION: Compared with the control group, after 24-hour intervention of 5-azacytidine, 5-azacytidine with 3, 6, 12, 24 μmol/L increased significantly cell proliferation without dose-dependence (P < 0.05). After 48-hour intervention of 5-azacytidine, 5-azacytidine with 6, 12, 24 μmol/L increased significantly cell proliferation without dose-dependence (P  < 0.05). After 72-hour intervention of 5-azacytidine, 5-azacytidine with 12, 24 μmol/L decreased significantly cell proliferation without dose-dependence (P < 0.05). After 5-day and 7-day intervention of 5-azacytidine, 5-azacytidine has no effect on cell proliferation (P > 0.05). Compared with the control group, after 48-hour intervention of 5-azacytidine, 5-azacytidine with 6, 12, 24 μmol/L increased significantly telomerase reverse transcriptase expression in bone marrow mesenchymal stem cells without dose-dependence (P  < 0.05). In certain concerntration and time, 5-azacytidine can increase proliferation and expression of telomerase reverse transcriptase in bone marrow mesenchymal stem cells.

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