Screening Study Of Differentially Expressed Micrornas In Primary And Recurrence Giant Cell Tumor Of Spine

ObjectiveUsing the microRNA microarray technology to analysis the differential expressed microRNAs in primary and recurrence tissue of giant cell tumor(GCT) of spine and followed with bioinformatics analysis to supply new evidences for realizing the molecular pathogenesis in occurrence, development and recurrence of giant cell tumor of spine.MethodsCollect 10 primary tumor tissue of GCT,10 recurrence tissue of GCT,5 normal bone tissue and 5 muscle tissue.Total RNA of tissues were extracted by Trizol and hybridized with the microarray.The slides were scanned and followed by data extraction.After statistical analysis we got the list of differential expressed microRNAs.Target gene prediction was made and their potential physiological functions were generated from the bioinformatic analysis.ResultsA list of differential expressed microRNAs was made by setting the hreshold of up/down regulated microRNAs as Foldchange>=1.5 and P value<=0.5. The differential expressed microRNAs in primary and recurrence tissue of GCT were as follow: 8 microRNAs were up-regulated——hsa-miR-22*, hsa-miR-1909, hsa-miR-218-2*,hsa-miRPlus-E1015,hsa-miRPlus-E1215,hsa-miR-29a*,hsa-miR-1323,hsa-miR-520a-5p;2 microRNAs were down-regulated——hsa-miR-944, hsa-miR-331-3p. Through the online target gene analysis tool,we found that hsa-miR-22* has 261 target genes, hsa-miR-1909 has 163 target genes, hsa-miR-218-2*has 233 target genes, hsa-miR-29a* has 340 target genes,hsa-miR-1323 has 460 target genes, hsa-miR-520a-5p has 649 target genes,hsa-miR-944 has 624 target genes, hsa-miR-331-3p has 390 target genes.Further more,the significant Gene Ontology(GO) classifications and Kyoto Encyclopedia Gene & Genomes(KEGG) pathways that were over–represented among these target genes were found out.And hsa-miR-1323, hsa-miR-520a-5p, hsa-miR-944,hsa-miR-331-3p were found being related with Pathways in cancer and VEGF(vascular endothelial growth factor) signaling pathway,which played important role in tumor formation. Conclusion1.MicroRNA microarray technology can be used for high-throughput screening of miRNA.2.A list of differential expressed microRNAs was made. The differential expressed microRNAs in primary and recurrence tissue of GCT were as follow: 8 microRNAs were up-regulated——hsa-miR-22*, hsa-miR-1909, hsa-miR-218-2*, hsa-miRPlus-E1015, hsa-miRPlus-E1215,hsa-miR-29a*,hsa-miR-1323,hsa-miR-520a-5p;2 microRNAs were down-regulated——hsa-miR-944, hsa-miR-331-3p.3. Through the online target gene analysis tool,we found target genes of the differential expressed microRNAs in primary and recurrence tissue of GCT.4.The significant Gene Ontology(GO) classifications and Kyoto Encyclopedia Gene & Genomes(KEGG) pathways that were over–represented among these target genes were found out.And hsa-miR-1323, hsa-miR-520a-5p, hsa-miR-944,hsa-miR-331-3p were found being related with Pathways in cancer and VEGF(vascular endothelial growth factor) signaling pathway,which played important role in tumor formation.5.The function of many miRNAs are not yet well understood including the differential expressed microRNAs in primary and recurrence tissue of GCT.Thus the practical application of miRNAs have not been carried out in clinical practice.The loss -of-function and gain -of-function of specific miRNA and their effect on the expression pattern of the encoding proteins of miRNA target genes will be a new direction.