解放军医学院学报
解放軍醫學院學報
해방군의학원학보
Academic Journal of Chinese Pla Medical School
2013年
10期
1063-1066,1100
,共5页
王涛%穆长征%王小梅%田鹤%陈修思
王濤%穆長徵%王小梅%田鶴%陳脩思
왕도%목장정%왕소매%전학%진수사
miRNA%NeuroD1%胰岛素分泌细胞%小鼠
miRNA%NeuroD1%胰島素分泌細胞%小鼠
miRNA%NeuroD1%이도소분비세포%소서
miRNA%NeuroD1%insulin-secreting cells%mice
目的：实现骨髓间充质干细胞(bone marrow mesenchymal stem cells，BMSCs)向胰岛素分泌细胞(insulin-producing cells，IPCs)的诱导分化并验证分化过程中miR-217靶向调控NeuroD1基因表达。方法分离培养BMSCs，应用大鼠胰腺损伤提取物(rat pancreatic extraction，RPE)将其诱导分化为IPCs。采用靶基因预测软件miRanda和TargetScan对miR-217和NeuroD1基因的靶向匹配关系进行预测并通过双荧光素酶报告基因系统鉴定。qRT-PCR检测诱导分化过程中miR-217和NeuroD1的表达。结果诱导分化后的细胞双硫腙染色呈猩红色，免疫荧光化学显示有胰岛素表达。生物信息学方法预测发现miR-217和NeuroD1基因二者匹配良好，通过双荧光素酶报告基因系统检测发现miR-217能结合到NeuroD1 mRNA的3'UTR并有效抑制其表达。qRT-PCR检测结果表明，miR-217的表达水平与NeuroD1表达呈负相关。结论 miR-217能调控IPCs诱导分化过程中NeuroD1的表达。
目的：實現骨髓間充質榦細胞(bone marrow mesenchymal stem cells，BMSCs)嚮胰島素分泌細胞(insulin-producing cells，IPCs)的誘導分化併驗證分化過程中miR-217靶嚮調控NeuroD1基因錶達。方法分離培養BMSCs，應用大鼠胰腺損傷提取物(rat pancreatic extraction，RPE)將其誘導分化為IPCs。採用靶基因預測軟件miRanda和TargetScan對miR-217和NeuroD1基因的靶嚮匹配關繫進行預測併通過雙熒光素酶報告基因繫統鑒定。qRT-PCR檢測誘導分化過程中miR-217和NeuroD1的錶達。結果誘導分化後的細胞雙硫腙染色呈猩紅色，免疫熒光化學顯示有胰島素錶達。生物信息學方法預測髮現miR-217和NeuroD1基因二者匹配良好，通過雙熒光素酶報告基因繫統檢測髮現miR-217能結閤到NeuroD1 mRNA的3'UTR併有效抑製其錶達。qRT-PCR檢測結果錶明，miR-217的錶達水平與NeuroD1錶達呈負相關。結論 miR-217能調控IPCs誘導分化過程中NeuroD1的錶達。
목적：실현골수간충질간세포(bone marrow mesenchymal stem cells，BMSCs)향이도소분비세포(insulin-producing cells，IPCs)적유도분화병험증분화과정중miR-217파향조공NeuroD1기인표체。방법분리배양BMSCs，응용대서이선손상제취물(rat pancreatic extraction，RPE)장기유도분화위IPCs。채용파기인예측연건miRanda화TargetScan대miR-217화NeuroD1기인적파향필배관계진행예측병통과쌍형광소매보고기인계통감정。qRT-PCR검측유도분화과정중miR-217화NeuroD1적표체。결과유도분화후적세포쌍류종염색정성홍색，면역형광화학현시유이도소표체。생물신식학방법예측발현miR-217화NeuroD1기인이자필배량호，통과쌍형광소매보고기인계통검측발현miR-217능결합도NeuroD1 mRNA적3'UTR병유효억제기표체。qRT-PCR검측결과표명，miR-217적표체수평여NeuroD1표체정부상관。결론 miR-217능조공IPCs유도분화과정중NeuroD1적표체。
Objective To identify the role of miR-217 in target regulation of NeuroD1 gene expression when bone marrow mesenchymal stem cells (BMSCs) are differentiated into insulin-secreting cells (IPCs). Methods BMSCs were isolated, cultured and differentiated into IPCs using rat pancreatic extraction (RPE). The relation between miR-217 and NeuroD1 target match was predicted using miRanda and TargetScan, and identified with the dual luciferase report system. Expression of miR-217 and NeuroD1 was detected by qRT-PCR when the BMSCs were differentiated into IPCs. Results The differentiated IPCs were stained scarlet with DTZ. Immunofluorescence cytochemistry showed the expression of insulin. The bioinformatics method showed that the miR-217 and NeuroD1 were well matched. Dual luciferase reporter system revealed that miR-217 was bound to 3'UTR of NeuroD1 mRNA and inhibited its expression. The qRT-PCR displayed that the expression of miR-217 was negatively correlated with that of NeuroD1 mRNA. Conclusion miR-217 can regulate the expression of NeuroD1 in the differentiation of IPC.
