人牙源性多能干细胞重编程前后微小RNAs差异表达谱系分析  

Characterization of microRNAs profiles of induced pluripotent stem cells reprogrammed from human dental pulp stem cells and stem cells from apical papilla

作　　者：谭小兵[1] 戴青原[2] 

机构地区：[1]云南省第一人民医院口腔内科 [2]昆明医科大学第一附属医院心内科,昆明650032

出　　处：《华西口腔医学杂志》2017年第3期269-274,共6页West China Journal of Stomatology

基　　金：国家自然科学基金(81360161);云南省教育厅基金(2-015Y153)~~

摘　　要：目的对比研究两种人牙源性多能干细胞重编程前后微小RNAs(mi RNAs)差异表达,交集分析、筛选特异性mi RNAs。方法利用仙台病毒将人牙髓干细胞(DPSCs)和根尖乳头干细胞(SCAP)重编程为诱导性多潜能干细胞(i PSCs),提取总RNA,mi RNAs标记、杂交,扫描芯片、读取图像,筛选差异表达mi RNAs,交集分析。结果人DPSCs和SCAP均可重编程为i PSCs。mi RNAs芯片分析结果显示人DPSCs重编程后有68个差异表达mi RNAs(倍数>10),其中37个表达上调,31个表达下调;人SCAP重编程后有107个差异表达mi RNAs(倍数>10),其中68个表达上调,39个表达下调。二者取交集,均上调的有mi R-302e,下调的有mi R-29b-3p、mi R-181b-5p、mi R-4328、mi R-22-5p、mi R-145-5p、mi R-4324、let-7b-5p、mi R-181a-5p、mi R-27b-3p(倍数>10)。结论人DPSCs和SCAP重编程为i PSCs过程中有多种mi RNAs参与,多数与细胞周期、上皮-间充质转化、转化生长因子β信号通路等相关。Objective To compare characterization of microRNAs （miRNAs） expression profiles of induced pluripotent stem cells （iPSCs） reprogrammed from human dental pulp stem cells （DPSCs） and stem cells from apical papilla （SCAP） and screen-specific microRNA. Methods Human DPSCs and SCAP were reprogrammed into iPSCs using a Sendai virus vector. Total RNA of human DPSCs-iPSCs and SCAP-iPSCs were extracted, miRNAs were labeled and hybridized. Slides were scanned, and images were imported into GenePix Pro 6.0 for grid alignment and data extraction. Significant differentially expressed miRNAs between the two groups were identified using fold change and P-value and were analyzed. Results Both human DPSCs and SCAP were successfully reprogrammed into iPSCs. Among miRNA genes analyzed by miRNA microarray, 68 were differentially expressed by more than 10-fold in DPSCs-iPSCs; 37 of these genes were up-regulated, and 31 were down-regulated. In SCAP-iPSCs, 107 genes were differentially expressed by more than 10-fold; 68 were up-regulated, and 39 were down-regnlated. In both cells, only miR-302e was up-regulated, whereas 9 miRNAs were down-regulated： miR-29b- 3p, miR-181b-5p, miR-4328, miR-22-5p, miR-145-5p, miR-4324, let-7b-5p, miR-181a-5p, and miR-27b-3p. Conclusion Multiple miRNAs participated in reprogramming of human DPSCs and SCAP into iPSCs. Most miRNAs are related to cell cycle, transforming growth factor-β signaling pathways and epithelial-mesenchymal transition.

关 键 词：诱导性多潜能干细胞 牙髓干细胞 根尖乳头干细胞 重编程 微小RNAS 

分 类 号：R781[医药卫生—口腔医学]