龟甲中具有潜在药理活性的miRNA初步筛选

Study on Preliminary Screening of miRNAs with Potential Pharmacological Activity in Plastrum Testudinis

目的:对龟甲中存在的miRNA进行提取并测序,并对具有潜在药理活性者进行初步筛选。方法:对生龟甲进行miRNA转录组测序,选出count值超过10 000的miRNA,并利用实时荧光定量PCR技术在龟甲、乌龟背甲、黄喉拟水龟背甲RNA中分别测定其拷贝数。结果:转录组数据中共筛选得到3条miRNA,其中1条为已知的xtr-miR-22;2条为未知,分别命名为NW-006642957-1-500和NW-006622633-1-195。荧光定量PCR结果显示xtr-miR-22在龟甲及乌龟背甲中拷贝数明显高于同属动物黄喉拟水龟,且其靶基因预测结果显示其能够靶向调控PML基因的表达。结论:生龟甲在炮制加工后存在未降解的miRNA,其中xtr-miR-22具有治疗早幼粒细胞白血病的潜在药理活性。

Objective:To extract and sequence the miRNA in Plastrum Testudinis,and screen preliminarily the potential pharmacological activity.Methods:Transcriptome analyses were carried out on Plastrum Testudinis,and the miRNAs which have counts of more than 10 000 were selected.The copy number of these miRNAs in Plastrum Testudinis,Chinemys reevesii,Mauremys mutica were measured respectively by qRT-PCR.Results:3 miRNAs were selected from the transcriptome data:xtr-miR-22,NW-006642957-1-500 and NW-006622633-1-195.The results of qRT-PCR showed that the copy numbers of xtr-miR-22 in Plastrum Testudinis,Chinemys reevesii were significantly higher than that of the counterpart in Mauremys mutica.The target gene prediction confirmed that xtr-miR-22 target expression of PML Gene.Conclusion:The miRNAs do exist in Plastrum Testudinis after deep-processing,and xtr-miR-22 which was discovered in Plastrum Testudinis have potential pharmacological activity to treat promyelocytic leukemia.