华西口腔医学杂志2019，Vol.37Issue(1)：25-30,6.DOI:10.7518/hxkq.2019.01.005
腺病毒介导的人端粒酶反转录酶牙周膜细胞系的建立
Construction of human telomerase reverse transcriptase periodontal ligament cell line mediated by adenovirus
摘要
Abstract
Objective This study aims to establish an effective and stable periodontal ligament cell line stably expressing human telomerase reverse transcriptase (hTERT) gene by using the adenovirus method.Methods Polymerase chain reaction (PCR) was used to amplify the full length of hTERT gene to construct recombinant adenovirus plasmid pAd-pshuttle-cmvhTERT.Packaged adenovirus particles were used for infection of human periodontal ligament cells.The expression levels of h TERT and osteogenic genes, such as alkaline phosphatase, Runt-related transcription factor 2, bone sialoprotein, osteocalcin, osteopontin, and collagen Ⅰ mRNA, were detected by quantitative real-time PCR (qRT-PCR).The ability of osteogenic differentiation was observed by alizarin red staining, and the cell proliferation was determined by CCK-8.Results Adenovirus particles containing the hTERT gene were successfully constructed and infected with periodontal ligament cells.The infected cells were similar to normal periodontal ligament cells.The qRT-PCR results showed that hTERT and osteogenesis-associated genes were highly expressed in the periodontal ligament cell lines constructed by adenoviruses.Alizarin red staining showed that the periodontal ligament cell line had strong osteogenic differentiation capability.CCK-8 showed that the periodontal ligament cell line had strong proliferation capability.Conclusion The human periodontal ligament cell line with high efficiency and stable expression of hTERT was established by the adenovirus method, thereby providing an ideal cell line for studying the mechanism of periodontal regeneration.关键词
牙周膜细胞/人端粒酶反转录酶/腺病毒/稳定表达/成骨分化Key words
periodontal ligament cells/human telomerase reverse transcriptase/adenovirus/stable expression/osteogenesis分类
医药卫生引用本文复制引用
孙贵军,秦晓东,王世琴,胡慧珍,陆笑颜,金佳佳,吴生荣,何祥一..腺病毒介导的人端粒酶反转录酶牙周膜细胞系的建立[J].华西口腔医学杂志,2019,37(1):25-30,6.基金项目
甘肃省自然科学基金 (17JR5RA217) （17JR5RA217）
