Expression of wingless-type MMTV integration site family member 10A in human dental pulp tissues and dental pulp cells
10.3760/cma.j.issn.1002-0098.2013.s1.008
- VernacularTitle:无翅型MMTV整合位点家族成员10A在人牙髓组织和细胞中的表达
- Author:
Qing-Ru GUO
1
;
Hua TIAN
;
Ping L(U)
;
Zhi-Chun ZHANG
;
Xue-Jun GAO
Author Information
1. 100081,北京大学口腔医学院·口腔医院牙体牙髓科
- Keywords:
Wnt proteins;
Cell proliferation;
Cell differentiation;
Dental pulp cells
- From:
Chinese Journal of Stomatology
2013;48(z1):34-39
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate wingless-type MMTV integration site family,member 10A (WNT10A) expression in human dental pulp tissues and human dental pulp cells (HDPC).Methods Human premolars or wisdom teeth were obtained fron healthy individuals extracted for orthodontic purpose.The expression of WNT10A and the molecules of Wnt signaling pathway in human dental pulp tissues were analyzed by immunohistochemistry and semi-quantitative reverse transcription-PCR(RT-PCR).HDPC were primarily cultured from pulp tissues of healthy premolars or wisdom teeth and passaged cells.Their growth patterns were observed by microscopy.The expression levels of Wnt molecules and odontoblast-specific biomarkers during different cell passage were determined by semi-quantitative and real-time quantitative RT-PCR.Results The immunohistochemical analysis showed WNT10A was located in the cytoplasms of odontoblasts.WNT10A,axis inhibition protein 2(AXIN2),Dickkopf(DKK1),lymphoid enhancer factors 1 (LEF1),low density lipoprotein receptor-related protein 4 (LRP4),dentin sialophoshoprotein (DSPP),alkaline phosphatase(ALP) and osteocacin were detected in the pulp tissues by RT-PCR.HDPC reached confluence in 15 days.The 4th passage of HDPC proliferated actively,with clear cytoplasms and uniformly stretches.The 9th passage of HDPC tended to die.The Wnt signaling pathway downstream molecules were expressed in HDPC throughout the subculture.DSPP expression at mRNA level was detected peaking in the 4th passage(0.4178 ±0.0076).The expression levels of WNT10A in the 5th and the 9th passage(4.97 ±2.83,4.70 ±4.06) were significantly higher than in the 2th and the 4th passage (0.84 ± 0.66,0.70 ±0.45) (P ＜ 0.05).Conclusions Wnt signaling pathway mediated by WNT1 0A was activated in pulp tissues and HDPC,indicating that WNT10A could be involved in differentitation of odontoblasts and dentin formation.
