Role of transient receptor potential melastatin 7 in the proliferation of human dental pulp stem cells

doi:10.3969/j.issn.2095-4344.2013.19.012

Abstract

Abstract:

BACKGROUND: Transient receptor potential melastatin 7 channel is widely distributed in tissues and cells, and it is necessary for cell survival, proliferation and maintenance of magnesium ion balance.
OBJECTIVE: To investigate the role of transient receptor potential melastatin 7 channel in the proliferation of human dental pulp stem cells.
METHODS: Human dental pulp stem cells were isolated and cultured by collagenase digestion method, and were exposed for 72 hours to 2-aminoethoxydiphenyl borate (50 and 100 μmol/L), which was widely used as an inhibitor of transient receptor potential melastatin 7. The influences of 2-aminoethoxydiphenyl borate on transient receptor potential melastatin 7 protein level and proliferation of human dental pulp stem cells were detected with western blot assay and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method, respectively. Specific lentivirus vector was constructed to inhibit the expression of transient receptor potential melastatin 7, and lentivirus was applied to transfect human dental pulp stem cells. Reverse transcription-PCR and western blot for silencing efficacy analysis were also performed. Changes of cell proliferation after specific silencing transient receptor potential melastatin 7 was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay at 1, 3, 5, and 7 days after culture.
RESULTS AND CONCLUSION: The proliferation of human dental pulp stem cells and protein expression of transient receptor potential melastatin 7 were significantly inhibited by 2-aminoethoxydiphenyl borate at 50 and 100 μmol/L (P < 0.01). After specific silencing transient receptor potential melastatin 7, the proliferation capacity of human dental pulp stem cells was significantly decreased at different time poinrs (P < 0.01). Transient receptor potential melastatin 7 plays an important role in maintaining proliferation ability of human dental pulp stem cells.

Key words: stem cells, stem cell culture and differentiation, transient receptor potential melastatin 7, human dental pulp stem cells, short-hairpin RNA, lentivirus, 2-aminoethoxydiphenyl borate, proliferation, provincial grants-supported paper, stem cell photographs-containing paper

CLC Number:

R394.2

Cui Li, Xu Shuai-mei, Wu Bu-ling. Role of transient receptor potential melastatin 7 in the proliferation of human dental pulp stem cells[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(19): 3495-3502.

Figures/Tables 6

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