摘要
目的:研究细胞外基质磷酸化糖蛋白(MEPE)基因启动子在小鼠前成骨细胞中表达的调控机制。方法:利用软件分析小鼠的MEPE基因启动子,得到5条启动子候选序列后,设计引物进行不同长度MEPE基因启动子的构建,并分别将其转染小鼠前成骨细胞;然后利用双荧光素酶基因检测报告系统分析不同长度MEPE基因启动子在前成骨细胞中的转录活性,并同时观察BPM2对小鼠前骨细胞中不同长度MEPE基因启动子转录活性的影响及其时间效应。结果:成功构建了P-78^+66、P-140^+66、P-300^+66、P-500^+66、P-1 081^+66的MEPE基因启动子;分别转染小鼠前成骨细胞后,不同长度MEPE基因启动子的活性两两相比均有显著性差异(P<0.05),其中以P-500^+66的活性最强,推测-140^-500区域可能为MEPE转录表达的调控元件主要结合部位;BMP2作用于不同长度MEPE基因启动子后检测发现,BMP2在启动子-140^-500区域内可明显上调MEPE基因的转录活性(P<0.05)。结论:成功构建了MEPE基因启动子,并发现BMP2调控MEPE基因启动子的主要区域在-140^-500区域。
AIM： To study the molecular mechanisms of MEPE gene promoter expression in preosteoblasts in mouse. METHODS： MEPE gene promoter of mouse preosteoblasts was analysed by software（http：//www, fruitfly. org/seq_tools/promoter, html）, five candidate sequences were obtained, the primers for the construction of different lengths of MEPE gene promoters were designed; dual luciferase reporter assay was applied to analysis the effects for transcription activity of MEPE gene promoters with different lengths. The effects of BMP2 on the transcription activity of MEPE gene promoters in preosteoblasts were observed. RESULTS ： MEPE gene promoters with the lengths of P - 78 - ＋66, P-140 - ＋66, P-300 - ＋66, P-500 - ＋66 and P-1081- ＋66 were obtained. Double luciferase genetic testing report system analysis showed that the transcription activity of different lengths MEPE gene promoters was differ- ent（P 〈0.05） . The transcription activity of MEPE gene promoter was significantly upregulated in the area of - 140 - -500 after BMP2 treatment. CONCLUSION： The location of MEPE gene promoter in mouse preosteoblasts was re- constructed. BMP2 may regulate the MEPE gene promoter at - 140 - -500 area.
出处
《牙体牙髓牙周病学杂志》
CAS
2015年第9期519-523,共5页
Chinese Journal of Conservative Dentistry
基金
国家自然科学基金(81441107)
山东省自然科学基金(ZR2012HQ036)
山东省教育厅(J12LL51)
