获得性血友病A患者血清miRNAs差异表达谱及功能miRNAs的初筛

Differential expression profile of serum miRNAs and primary screening of functional miRNAs in acquired hemophilia A patients

目的 建立获得性血友病A(AHA)患者血清miRNAs差异表达谱,并探究潜在功能miRNAs分子。方法 收集13例AHA患者和13例健康对照的血清样本、人口统计学特征、APTT、FⅧ:C、FⅧ:Ab以及基础疾病情况,利用微列阵芯片技术分析其中3例AHA患者和3例健康对照血清样本,筛选差异表达miRNAs,并建立差异表达谱。在另外20例参试者(AHA和健康对照各10例)中,利用荧光定量PCR(qRT-PCR)技术分析验证差异表达miRNAs,筛选出潜在功能miRNAs。结果 微列阵芯片分析结果表明,有23个miRNAs在AHA患者和健康对照血清中表达水平存在显著差异(P<0.05)。实时荧光定量PCR技术(qRT-PCR)分析结果表明,与健康对照相比,AHA患者血清中4个miRNAs(miR-206、miR-144-5p、miR-374b-5p、miR-432-3p)显著上调(P<0.05),1个miRNA(miR-92a-3p)显著下调(P<0.05),其余18个miRNAs表达水平的差异不明显(P>0.05)。筛选出的5个miRNAs中,miR-206和miR-374b-5p与患者FⅧ:Ab显著正相关(P<0.05),而miR-92a-3p则与FⅧ:Ab显著负相关(P<0.05)。此外,miR-206和miR-92a-3p与患者罹患风湿性疾病、产后并发症、恶性肿瘤类基础疾病也存在显著相关性(P<0.05)。结论 本研究基于AHA患者血清建立了miRNAs差异表达谱,并筛选得到5个显著差异表达的miRNAs,为未来预防、诊断和治疗AHA提供了候选分子。

Objective To construct the differential expression profile of serum miRNAs in acquired hemophilia A(AHA)patients,and screen the potential functional miRNAs.Methods The serum samples,demographic characteristics,APTT,FⅧ:C,FⅧ:Ab and basic diseases from 13 AHA patients and 13 healthy controls were collected.The differentially expressed miRNAs were screened in serum from 3 AHA patients and 3 healthy controls by microarray chip technology,and the differential expression profile was construct-ed.In addition,quantitative real-time PCR(qRT-PCR)was used to verify the differentially expressed miRNAs in the remaining 20 participants(10 cases of AHA and 10 cases of healthy controls),and screen the potential functional miRNAs.Results The results of chip showed that 23 miRNAs were differentially expressed in serum of AHA compared to healthy controls(P<0.05).The qRT-PCR results showed that four miRNAs(miR-206,miR-144-5p,miR-374b-5p,miR-432-3p)were highly expressed in AHA patients compared with healthy controls(P<0.05),while one miRNA(miR-92a-3p)was downregulated in AHA patients compared with healthy controls(P<0.05).The remaining 18 miRNAs showed no significantly differential expression between AHA patients and healthy controls(P>0.05).Among the five differentially expressed miRNAs,miR-206 and miR-374b-5p were positively correlated with FⅧ:Ab(P<0.05),while miR-92a-3p was negatively correlated with FⅧ:Ab in AHA patients(P<0.05).Additionally,miR-206 and miR-92a-3p were associated with basic diseases,such as rheumatic diseases,postpartum complications and malignant tumors in AHA patients(P<0.05).Conclusion In this study,the differential expression profile of miRNAs is established based on the serum of AHA patients,and five miRNAs with significant differential expression are screened,providing candidate molecules for the prevention,diagnosis and treatment of AHA.