牙周膜细胞内miR-23a、miR-30a发挥靶向调控作用的基因筛选  

Screening of Genes Targeting the Regulation of MicroRNA-23a and MicroRNA-30a in Periodontal Ligament Cells

作　　者：王献刚[1] 郝志红[1] 袁清敏 冯保静[1] Wang Xiangang;Hao Zhihong;Yuan Qingmin;Feng Baojing(Department of Stomatology of Anyang Sixth People's Hospital(Stomatological Hospital),Anyang 455000,China)

机构地区：[1]河南省安阳市第六人民医院(口腔医院)口腔修复科

出　　处：《四川解剖学杂志》2019年第4期1-4,共4页Sichuan Journal of Anatomy

摘　　要：目的筛选测定牙周膜细胞内miR-23a、miR-30a发挥靶向调控作用的基因。方法采用miRanda、PicTar及TargetSan三个靶基因预测软件预测miR-23a、miR-30a的候选靶基因;通过体外炎性微环境模拟检测及炎症牙周膜细胞的miR-23a、miR-30a表达水平干预,采用实时定量PCR仪检测miR-23a、miR-30a及其候选靶基因表达情况,筛选miR-23a、miR-30a可能的靶基因;并采用免疫印迹法对靶蛋白进行进一步鉴定。结果预测软件提示,miR-23a、miR-30a有Runx2、CAMTA1、STAT1、STATB1、CNTTBIP、STAT5B、PPPAR七个共同的候选靶基因;实时定量PCR检测提示,STAT5B与Runx2可能是miR-23a、miR-30a的靶基因;免疫印迹检测提示STAT5B与Runx2蛋白表达水平变化趋势与实时定量PCR检测结果具有一致性,提示STAT5B可能是miR-23a的靶基因,Runx2可能是miR-30a的靶基因。结论 miR-23a、miR-30a均参与了炎性微环境下牙周膜细胞的生物学活性表达,且STAT5B、Runx2分别可能是miR-23a、miR-30a的靶基因。Objective This study aimed to screen and determine the genes targeting the expression of microRNA-23a and microRNA-30a in periodontal ligament cells. Methods The candidate target genes were predicted by three software of microRNAanda, PicTar and TargetSan. The expression levels of microRNA23a,microRNA30a and their possible candidate target genes of inflammatory periodontal ligament cells were detected by real-time quantitative PCR under in vitro inflammatory microenvironment simulation and the intervention. The expression of microRNA23a, microRNA30a and their candidate target proteins were screened and identified by Western blotting. Results The predictive software indicated that there were seven common candidate target genes Runx2,CAMTA1,STAT1,STATB1,CNTTBIP,STAT B and PPPAR for microRNA-23a and microRNA-30a. Real-time quantitative PCR indicated that STAT5B and Runx2 might be the target genes of microRNA-23a and microRNA-30a. Western blotting showed that the expression of STAT5B and Runx2 was consistent with the results of real-time quantitative PCR, suggesting that STAT5B and Runx2 might be the target genes of microRNA-23a and microRNA-30a, respectively. Conclusion MicroRNA-23a and microRNA-30a are involved in the biological activity changes of periodontal ligament cells of inflammatory microenvironment, and STAT5B and Runx2 have targeted regulatory relationship with microRNAs-23a and microRNAs-30a respectively.

关 键 词：牙周膜细胞 miR-23a miR-30a 靶向调控 基因筛选 

分 类 号：R781[医药卫生—口腔医学]