Screen Differentially Expressed MicroRNAs In Peripheral Blood Cells Of Primary Immune Thrombocytopenia And Systemic Lupus Erythematosus Patients

Objective：By comparing microRNAs in primary immune thrombocytopenia (ITP)and systemic lupus erythematosus (SLE) patients with differential expressionin peripheral blood cells, found that patients with ITP is specific miRNAexpression, can provide early in the disease to distinguish ITP from SLE toprovide corresponding basis.Methods：Primary immune thrombocytopenia patients and patients withsystemic lupus erythematosus all five, each peripheral venous EDTAanticoagulant2ml, Ficoll gradient centrifugation separation of peripheralblood mononuclear cells, to join the Trizol cracking, respectively a group ofpatients with ITP and SLE patients group of samples of mixture, the extractionof total RNA. Use gel electrophoresis to verify integrity of RNA and Nanodropspectrophotometer determination of RNA concentration and purity. Use miRCURYTMHy3/Hy5kit tag of micrornas. Application miRCURYTMLNA Array (v.16.0)kit for Hy3tag samples of miRNAs. With Axon GenePix4000B miRNAs chip scannerscan, GenePix Pro6.0software (Axon) to analyze the miRNAs signal strength.The miRNAs data (signal strength50or higher), standardization to the medianto express differences of miRNAs using MEV software (v4.6, TIGR) forhierarchical clustering analysis. Pick up purpose of miRNAs from the expressiondifferences of miRNAs, RT-PCR results show.Results：There were121differential expressed miRNAs, among which55onlyexpressed in primary immune thrombocytopenia group while56only expressed insystemic lupus erythematosus group.10miRNAs expressed opposite in the ITPgroup and the SLE group,of which4up-regulated in the ITP group and down-regulated in the SLE group,while6up-regulated in the ITP group and down- regulated in the SLE group. Hierarchical cluster analysis show that there arespecific miRNAs expression spectrum of ITP. RT-PCR detection results and theanalysis result is consistent with the chip.Conclusions：1. Using the chip technology compared ITP patients with SLE patients withperipheral blood cells miRNAs expression spectrum in mixed samplespecimens,found121differential expressed miRNAs, among which55onlyexpressed in primary immune thrombocytopenia group while56only expressed insystemic lupus erythematosus group.10miRNAs expressed opposite in the ITPgroup and the SLE group,of which4up-regulated in the ITP group and down-regulated in the SLE group,while6up-regulated in the ITP group and down-regulated in the SLE group. Hierarchical cluster analysis show that there arespecific miRNAs expression in patients with ITP, further RT-PCR test verifythe reliability of chip results.2. ITP and SLE peripheral blood cells of each specific miRNAs expressionabnormalities, prompt the miRNAs expression changes may be involved in thepathogenesis of ITP.3. MicroRNAs diagnostic marker of biology, is expected to become the ITPfor early in the disease to distinguish ITP from SLE to provide correspondingbasis.4. Provide the basis for targeted treatment of ITP.