慢病毒介导cbfα1基因感染对人牙龈成纤维细胞成骨特性的影响

›› 2020, Vol. 40 ›› Issue (8): 682-687.

慢病毒介导cbfα1基因感染对人牙龈成纤维细胞成骨特性的影响

陈晓霞¹,颜世果²,孙钦峰²,李玉兰¹

1. 陕西省康复医院
2. 山东大学口腔医学院

收稿日期:2020-02-20 修回日期:2020-04-28 出版日期:2020-08-29 发布日期:2020-08-28
通讯作者: 李玉兰 E-mail:282564941@qq.com
基金资助:
山东省科技发展计划项目

慢病毒介导cbfα1基因感染对人牙龈成纤维细胞成骨特性的影响

Received:2020-02-20 Revised:2020-04-28 Online:2020-08-29 Published:2020-08-28

摘要/Abstract

摘要： ［摘要］　目的　研究慢病毒介导cbfα1基因感染对人牙龈成纤维细胞（hGFs）成骨特性的影响。方法：构建携带目的基因的慢病毒表达载体pLenti6.3-cbfα1-Ires-EGFP (pL-cbfα1)（实验组），通过酶切、测序验证其构建成功，利用293T细胞包装病毒，并测定病毒浓度，将病毒液感染hGFs，pLenti6.3-Ires-EGFP (pL-E)为空载体组，未被感染的正常hGFs为空白对照组，通过荧光实时定量PCR检测cbfα1的mRNA的表达，通过细胞免疫化学染色检测cbfα1蛋白的表达；通过荧光实时定量PCR检测各组成骨相关基因骨涎蛋白（BSP）、骨桥蛋白（OPN）、I型胶原（ColI）、骨钙蛋白（OC）的表达。结果：本实验慢病毒表达载体能够成功感染人牙龈成纤维细胞，并检测到cbfα1基因mRNA、蛋白的表达；实验组中，BSP、OPN、ColI、OC的表达明显高于对照组和空载体组（P<0.05）。结论：外源性基因cbfα1能够在人牙龈成纤维细胞中表达，并促进其向成骨细胞方向转化，此实验为进一步研究该基因及种子细胞在牙周组织再生工程中的作用奠定了基础。

关键词: cbfα1, 慢病毒, 人牙龈成纤维细胞

Abstract: Abstract：　Objective To study the effects of cbfα1 gene transfection on human gingival fibroblasts(hGFs). Methods To construct Lentivirus expression vector with the target gene cbfα1,and its construction was verified by enzyme digestion and sequencing. 293T cells were used to package the virus and the virus concentration was determined. The cbfα1 gene was transfected into human gingival fibroblasts by lentivirus. The hGFs were divided into three groups: Lentivirus plenti6.3-cbfα1- Ires-EGFP (pl-cbfα1) (experimental group), plenti6.3- Ires-EGFP (pl-E) (empty vector group), and the untransfected normal hGFs group. The expression of cbfα1 gene were determined by fluorescence real-time quantitative PCR and immunohistochemistry. Expressions of osteogenic genes BSP, OPN, ColI and OC in each group were detected by fluorescence real-time quantitative PCR. Results The results showed that the lentivirus with aim gene was able to infect human gingival fibroblasts successfully, and cbfα1 mRNA and protein were synthesized in the gingival fibroblasts. It was also found that the expressions of osteogenic genes BSP, OPN, ColI and OC in experimental group were significantly higher than those in the control groups(P<0.05). Conclusion The exogenous gene cbfα1 can be expressed in human gingival fibroblasts and promote its transformation to osteoblasts. This study was laying a foundation for further research on the role of this gene and seed cells in periodontal tissue regeneration engineering.

Key words: cbfα1, lentivirus, hGFs

中图分类号:

R349.64

陈晓霞颜世果孙钦峰李玉兰. 慢病毒介导cbfα1基因感染对人牙龈成纤维细胞成骨特性的影响[J]. 口腔医学, 2020, 40(8): 682-687.

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慢病毒介导cbfα1基因感染对人牙龈成纤维细胞成骨特性的影响

慢病毒介导cbfα1基因感染对人牙龈成纤维细胞成骨特性的影响

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