目的：通过构建Glil基因腺病毒载体,以此上调人牙周膜干细胞（ PDLSCs）中Glil基因的表达,从而探讨其对人PDLSCs增殖及成骨分化的影响。方法将Glil目的基因亚克隆至腺病毒载体,并将含目的基因的病毒转染PDLSCs细胞。 CCK-8法检测过表达Glil对PDLSCs增殖的影响,利用Western blot检测PDLSCs中Glil的表达水平及其成骨相关标志物ALP、Runx2的表达。结果成功构建了过表达Glil目的基因的腺病毒载体并转染PDLSCs,过表达Glil的PDLSCs与空载体组比较,增殖速率减慢（P=0.003）,Western blot检测结果显示PDLSCs过表达Glil目的基因后可以高表达成骨分化标记蛋白ALP、Runx2,差异具有统计学意义（P〈0.05）。结论过表达Glil基因会导致PDLSCs的增殖减慢,成骨相关标志物增加,说明Glil基因能使PDLSCs成骨分化能力增强。
Objective To up-regulate the expression of Glil gene in periodontal ligament stem cells （ PDLSCs） and to explore the effect of Glil gene on PDLSCs proliferation and osteogenesis differentiation by establishing Glil gene adenovirus vectors. Methods Subcloned Glil to viral backbone vector Adtrack-CMV and transfered the established vector to 293T cells, which was to acquire the virus particles. Trans-fected aim cells,namely PDLSCs,with these virus. Detected its effect on PDLSCs proliferation with CCK-8 assay, and detected the expression of Glil and the bone-related markers ALP and Runx2 through Western blot. Results An adenovirus vector, which were over expressed Glil gene, was successfully constructed and transfected to PDLSCs. Compared with the empty vector group and normal group, the over expressed one had a much slower proliferation rate in CCK-8 assay （P=0. 003）. Western blot showed that ALP and Runx2 can be overexpressing os-teogenic differentiated after PDLSCs successfully transfected with the Glil gene. Conclusion Over expressing Glil gene would lead to a much slower proliferation rate in the PDLSCs and an increase of the bone-related markers. It is concluded that Glil can enhance the osteogenic dif-ferentiation capacity in PDLSCs.
Journal of Regional Anatomy and Operative Surgery
human periodontal ligament stem cells