移植肾慢性排斥反应的miRNAs差异表达研究  

Differential expression of microRNAs in transplanted kidney undergoing chronic rejection

作　　者：眭维国[1] 蓝慧娟[1] 陈怀周[1] 黄河[1] 晏强[1] 陈洁晶[1] 戴勇[1] 

机构地区：[1]解放军第181医院肾脏科,广西桂林541002

出　　处：《解放军医学杂志》2008年第12期1441-1443,共3页Medical Journal of Chinese People's Liberation Army

基　　金：广西自然科学基金资助项目(0832289)

摘　　要：目的通过microRNAs芯片技术,探讨microRNAs(miRNAs)在移植肾慢性排斥反应中的差异表达情况。方法纳入4例慢性排斥反应患者肾穿组织为实验组,3例病理检查正常的肾组织皮质为对照组。采用miRNA芯片筛选出差异表达显著的微小RNA,并用实时定量PCR技术验证芯片结果可靠性。结果紫外分光光度法和变性电泳法证明所提取样本总RNA质量良好,可用于后续的芯片实验。芯片实验结果显示CR组中出现差异表达显著的miRNAs 63个,其中表达显著上调35个,表达显著下调28个。其中有9个miRNAs集中在3个miRNAs基因簇位点:14q32.31,22q11.21,xq27.3。随机选择显著差异表达的miRNAs(hsa-miR-637,hsa-miR-648,hsa-miR-516-5p)进行实时定量PCR验证实验,其CR/NC结果(0.034,2.670,7.846)与芯片结果(0.035,2.660,7.857)相符,证明了芯片结果的可靠性。结论miRNAs在慢性排斥反应患者中出现显著差异表达,深入研究miRNAs在慢性排斥反应中的作用机制将为其预防和治疗提供新途径。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 chroniie 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 Wrizol 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 electmphoresis 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 dusters： 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.

关 键 词：微RNAS 微阵列分析 肾移植 移植物排斥 

分 类 号：R617[医药卫生—外科学]