Lin28过表达对人牙髓细胞增殖、分化及矿化能力的影响  

Effect of Lin28 overexpression on proliferation,differentiation and mineralization of human dental pulp cells

作　　者：董宁[1] 张田田[2] 郭青玉[1] 阮建平[2] DONG Ning;ZHANG Tiantian;GUO Qingyu;RUAN Jianping(Clinical Research Center of Shaanxi Province for Dental and Maxillofacial Diseases,Department of Pediatric Dentistry,College of Stomatology,Xi’an Jiaotong University,Xi’an 710004,China;Department of Preventive Dentistry,College of Stomatology,Xi’an Jiaotong University)

机构地区：[1]陕西省颅颌面精准医学研究重点实验室,西安交通大学口腔医学院儿童口腔科,西安710004 [2]西安交通大学口腔医学院口腔预防科,西安710004

出　　处：《山西医科大学学报》2019年第9期1293-1298,共6页Journal of Shanxi Medical University

基　　金：陕西省科技计划项目(2015SF-153)

摘　　要：目的 研究Lin28过表达对人牙髓细胞(HDPCs)增殖、分化及矿化能力的影响。方法 通过慢病毒载体构建Lin28稳定过表达牙髓细胞株,实验分组为:正常HDPC组(正常组)、转染空载体的HDPCs组(空载体组)及Lin28过表达组。经矿化诱导培养后,分别用CCK-8法、ATP活性测定、碱性磷酸酶(ALP)活性测定和茜素红钙染色法检测Lin28过表达对人牙髓细胞增殖、能量、分化和矿化能力的影响;qRT-PCR检测成牙本质相关基因(DMP-1、DSPP、OCN、OPN、ALP)的mRNA相对表达水平。结果 CCK-8法检测增殖活性结果显示Lin28过表达组的OD值明显高于正常组和空载体组,差异具有统计学意义( P <0.05);Lin28过表达牙髓细胞ATP及ALP活性较空载体组和正常组明显升高( P <0.05);Lin28过表达组的钙化结节形成数量显著超过正常组,同时Lin28过表达组的牙本质形成相关基因DMP-1、DSPP、OCN、OPN、ALP的mRNA相对表达水平较空载体组和正常组上调( P <0.05)。而空载体组和正常组间的差异没有明显的统计学意义( P <0.05)。结论 Lin28过表达可增强人牙髓细胞的增殖、分化及矿化能力,以及牙本质形成相关基因mRNA的相对表达水平。Objective To investigate the effect of Lin28 overexpression on the proliferation, differentiation and mineralization of human dental pulp cells. Methods Lin28 stable overexpression dental pulp cell line was constructed by lentivirus vector. The experiment included normal human dental pulp cells(HDPCs) group, empty vector transfection group and Lin28 overexpression group. After mineralization induction, the effects of Lin28 overexpression on proliferation, cellular energy, differentiation and mineralization abilities of HDPCs were detected by CCK-8, ATP activity, alkaline phosphatase activity and alizarin red calcium staining. The relative level of odontoblast-related genes(DMP-1, DSPP, OCN, OPN, ALP) mRNA expression was detected by qRT-PCR. Results The results of CCK-8 showed that OD value in Lin28 overexpression group was significantly higher than that in normal HDPCs group and empty vector transfection group( P <0.05). ATP and ALP activity in Lin28 overexpression group were significantly higher than those in empty vector group and normal HDPCs group( P <0.05). Meanwhile, the relative mRNA expression of DMP-1, DSPP, OCN, OPN and ALP was significantly higher in Lin28 overexpression group than in empty vector group and normal group( P <0.05). There was no significant difference in above mentioned indexes between empty vector group and normal group( P <0.05). Conclusion Lin28 might play an important positive regulatory role in the proliferation, differentiation and mineralization of dental pulp cells.

关 键 词：Lin28 牙髓细胞 ATP 增殖 分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]