HOXC6同型蛋白2对根尖牙乳头干细胞成骨/成牙分化功能的影响  

The role of homeobox C6 isoform 2 in the osteogenic and dentinogenic differentiation of stem cells from apical papilla

作　　者：张琛[1] 李文芝[1] 葛丽华[2] 范志朋[2] 

机构地区：[1]首都医科大学口腔医学院牙体牙髓科,北京100050 [2]首都医科大学口腔医学院口腔医学研究所,北京100050

出　　处：《北京口腔医学》2016年第4期181-185,共5页Beijing Journal of Stomatology

基　　金：国家自然科学基金(81200783);北京市卫生系统高层次卫生技术人才培养计划(2013-3-039)

摘　　要：目的探讨同源盒基因C6同型蛋白2对牙源性间充质细胞成骨/成牙分化的影响。方法成骨诱导培养基诱导人根尖牙乳头干细胞体外成骨/成牙分化;逆转录病毒转染构建过表达HOXC6-ISO2稳定转染牙乳头干细胞进行HOXC6-ISO2获得性功能研究;碱性磷酸酶活性实验检测成骨/成牙早期分化指标-碱性磷酸酶活性;茜素红染色及钙离子定量分析检测干细胞体外矿化能力;实时荧光定量反转录PCR检测成骨/成牙分化相关基因-Sp7转录因子、Runt相关转录因子2和骨钙素及同源盒基因C6下游基因胰岛素结合蛋白5的表达。结果结果显示过表达同源盒基因C6同型蛋白2明显抑制牙乳头干细胞的碱性磷酸酶活性;茜素红染色及钙离子定量分析结果显示过表达同源盒基因C6同型蛋白2明显抑制牙乳头干细胞体外矿化能力;实时荧光定量反转录PCR显示过表达同源盒基因C6同型蛋白明显抑制骨钙素的表达,并且抑制下游基因胰岛素结合蛋白5的表达。结论同源盒基因C6同型蛋白2通过抑制下游基因胰岛素结合蛋白5抑制牙乳头干细胞体外成骨/成牙分化的功能。Objective To investigate the role of homeobox C6 isoform 2 （HOXC6-ISO2） in the osteogenic and dentinogenic differentiation potential of stem cells from apical papilla （SCAPs）. Methods Osteogenic-inducing medium was used to induce the osteogenic and dentinogenic differentiation of SCAPs. Retroviral HA-HOXC6-ISO2 was used to establish the stable cell and over-express HOXC6-ISO2 for gain-of-function study. The early marker of osteogenic and dentinogenic differentiation-ALP activity was detected by alkaline phosphatase （ALP） activity assay. Alizarin-red staining and quantitative analysis of calcium were used to investigate the mineralization potentials of SCAPs in vitro. Real time RT-PCR was used to detect the osteogenesis and dentinogensis related genes expressions, Sp7 transcript factor （osterix, OSX）, runt- related transcription factor 2 （RUNX2） and osteocalcin （ OCN）, and the downstream target gene of HOXC6, insulin-like growth factor binding protein 5 （IGFBP5）. Results Over-expression of HOXC6-ISO2 inhibited ALPactivity and mineralization in SCAPs in vitro. Over-expression of HOXC6-ISO2 inhibited the osteogenesis and dentinogensis related genes expression of OCN in SCAPs. Over-expression of HOXC6-ISO2 also inhibited the downstream target gene, IGFBPS. Conclusion HOXC6-ISO2 repressed the osteogenic and dentinogenic differentiation potential of SCAPs via IGFBPS.

关 键 词：HOXC6 同型蛋白2 根尖牙乳头间充质干细胞 成骨分化 

分 类 号：R781[医药卫生—口腔医学]