Differential expression of microRNAs in transplanted kidney undergoing chronic rejection / 解放军医学杂志

ABSTRACT

Objective To investigate the differential expression of microRNAs(miRNAs) in transplanted kidney undergoing chronic rejection by the technique of RNA microarray.Methods Four biopsy specimens from transplanted kidney undergoing chronic rejection were harvested as test group(CR),and 3 biopsy specimens were obtained from normal renal cortex as normal control group(NC).Total RNA of each sample was extracted using Trizol reagent.miRNAs were isolated and differential expression of miRNAs were screened by miRNA array analysis.The results of miRNA array were validated by RT-PCR.The quantity and quality of all the RNA samples were checked by gel electrophoresis and absorbance at A260/280,respectively.Results It was confirmed that the isolated RNA was of appropriate quality.The results of miRNA array analysis showed that there were 63 differential expression miRNAs in CR group,of which 35 were up-regulated and 28 down-regulated.There were 9 differential expression miRNAs which distributed in 3 gene clusters: 14q32.31,22q11.21 and xq27.3.The miRNAs hsa-miR-637,hsa-miR-648 and hsa-miR-516-5p were randomly selected for relative quantification by real-time PCR.It was showed that the expression ratios of hsa-miR-637,hsa-miR-648 and hsa-miR-516-5p in AR/NC,when detected by RT-PCR,were 0.034,2.670 and 7.846,while the ratios were 0.035,2.660 and 7.857 when analyzed by miRNAs array.The results from two methods were not significantly different,so the method of miRNAs array was reliable.Conclusions It is notable that the differential expression of miRNAs existed in the transplanted kidney undergoing chronic rejection.miRNAs might be helpful in protecting the patients undergoing kidney transplantation against chronic rejection.