Zhang Liang, Yu Cuntao, Chang Qian, Luo Xinjin, Qiu Juntao, Liu Shen
State Key Laboratory of Cardiovascular Disease, Aortic Surgery Center, Fuwai Hospital, National Center for Cardiovascular Disease, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100037, P.R. China.
Biomed Rep. 2016 Oct;5(4):421-427. doi: 10.3892/br.2016.740. Epub 2016 Aug 9.
The aim of the present study was to compare the gene expression profiles in aortic dissection (AD) and healthy human aortic tissue samples by DNA microarray analysis in order to screen the differential genes. In total, five AD and four healthy aortic specimens were selected; the total RNA was extracted and reverse transcribed into cDNA and transcribed into aRNA, followed by microarray hybridization for analysis. Thereafter, the transcription levels of six differential genes, myosin light chain kinase (), polycystin 1, transient receptor potential channel interacting (), myosin heavy chain 11 (), superoxide dismutase 3, extracellular (), filamin A (), and transgelin (), screened from the expression profiles were quantitatively verified. Compared with the healthy aortic specimens, a total of 1,661 genes in the AD group demonstrated more than 2-fold differential expression, of which 997 genes were upregulated and 664 genes were downregulated. Thereafter, six AD-associated genes that showed downregulation in the microarray assay were selected for quantitatively verifying the gene transcription level using reverse transcription-quantitative polymerase chain reaction (RT-qPCR), which confirmed their downregulation compared with the healthy aortic tissue genes; of the six genes, the expression levels of , , , and were significantly downregulated (P<0.05), while the expression of was not significantly downregulated (P>0.05). Thus, whole genome microarray may be used to screen differentially expressed genes between AD and healthy aortic tissues. When used in combination with RT-qPCR validation, this method may provide novel strategies for investigating AD.
本研究的目的是通过DNA微阵列分析比较主动脉夹层(AD)与健康人主动脉组织样本中的基因表达谱,以筛选差异基因。总共选取了5个AD样本和4个健康主动脉样本;提取总RNA并逆转录为cDNA,再转录为aRNA,随后进行微阵列杂交分析。此后,对从表达谱中筛选出的6个差异基因,即肌球蛋白轻链激酶()、多囊蛋白1、瞬时受体电位通道相互作用蛋白()、肌球蛋白重链11()、细胞外超氧化物歧化酶3()、细丝蛋白A()和原肌球蛋白()的转录水平进行了定量验证。与健康主动脉样本相比,AD组共有1661个基因表现出超过2倍的差异表达,其中997个基因上调,664个基因下调。此后,选择在微阵列分析中显示下调的6个与AD相关的基因,使用逆转录-定量聚合酶链反应(RT-qPCR)定量验证基因转录水平,结果证实与健康主动脉组织基因相比它们下调;在这6个基因中,、、、和的表达水平显著下调(P<0.05),而的表达下调不显著(P>0.05)。因此,全基因组微阵列可用于筛选AD与健康主动脉组织之间的差异表达基因。当与RT-qPCR验证结合使用时,该方法可为研究AD提供新策略。