A preliminary study on the feasibility of gene expression profile of rhesus monkey detected with human microarray
A preliminary study on the feasibility of gene expression profile of rhesus monkey detected with human microarray
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DOI：
10.1016/j.transproceed.2008.01.029
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发表时间：
2008-03-01
影响因子：
0.9
通讯作者：
Cheng, J. Q.
中科院分区：
文献类型：
--
作者：
Lu, Y. R.;Wang, L. N.;Cheng, J. Q.
目标。为了研究猪到人异种移植的结果，我们建立了猪到猴的移植模型。虽然猴子在生物学和生理学上与人类很接近，但这两个物种之间的基因差异尚不清楚。本研究旨在比较人类和恒河猴三种组织的基因表达。从肝脏、脾脏和外周血细胞中提取RNA样本，将其杂交到Illumina基因表达芯片上。选择检测到的信号大于1000且差异评分大于100的基因作为显著性结果。用Illumina软件对数据进行分析。采用半定量逆转录聚合酶链式反应（RT-PCR）分析证实mRNA表达水平。在每个基因芯片上测试的47293个转录本中，有6000多个基因在三种组织中表达。检测到的基因总数和相似比的规律与肝脏< PBC <脾脏相同。选择分析的136个iri相关基因、192个免疫相关基因和131个细胞周期相关基因的基因表达一致性率分别为82.35%、72.92%和77.10%。RT-PCR检测显示，RTN4、白细胞介素(IL)-1 β、nf - κ B1、IL-8和G0S2的mRNA表达水平与芯片检测结果相似。检测到的人与猴组织mRNA表达量的平均一致性为85.78%，表明人基因芯片可能为猴样本检测提供部分信息。因此，在猪到猴的移植模型中，猴子微阵列可以用来确定受体基因的表达。在解释实验结果时，必须考虑到人和猴子的基因差异。
Objectives. A pig-to-monkey transplant model was initiated to investigate the outcome of pig-to-human xenotransplantation. Though monkey is close to human in biology and physiology, the genetic differences between the two species remains unclear. This study sought to compare the gene expressions of three tissues from humans and rhesus monkey.Methods. RNA samples extracted from liver, spleen, and peripheral blood cells were hybridized onto Illumina gene expression microarray. Genes with detected signals greater than 1000 and diff-scores higher than 100 were selected as significant results. The data were analyzed with Illumina software. mRNA expression levels were confirmed by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) analysis.Results. Of the 47,293 transcripts tested on every gene chip, more than 6000 genes were expressed in three tissues. Total numbers of genes detected and the similarity ratios followed the same rule as liver < PBC < spleen. The 136 IRI-related genes, 192 immunological-related genes, and 131 cell cycle-related genes selected and analyzed showed gene expression concordance rates of 82.35%, 72.92%, and 77.10%, respectively. RT-PCR tests indicated similar mRNA expression levels of RTN4, interleukin (IL)-1 beta, NF-kappa B1, IL-8, and G0S2 to the results on chips.Conclusions. The detected mRNA expressions inhuman and monkey tissues showed an average consistency in 85.78%, indicating that a human microarray might provide a part of the information for monkey sample testing. Therefore, in pig-to-monkey transplant models, monkey microarray may be used to determine recipient gene expressions. The genetic difference between human and monkey must be taken into account in interpreting the experimental results.