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目的:利用基因芯片技术研究人主动脉夹层组织与正常主动脉组织基因表达谱的差异。方法:选取主动脉夹层病例标本5例,正常主动脉标本4例。提取总RNA,并反转录成cDNA、体外转录合成aRNA后与芯片进行杂交,对结果进行分析。同时对表达谱筛选出MYLK、PKD1、MYH11、SOD3、FLNA、TAGLN 6个差异基因进行基因转录水平的定量验证。结果:人主动脉夹层组与正常主动脉组比较基因表达差异倍数大于2的基因共有1 661个,其中有997个基因上调表达,664个基因下调表达。6个基因RT-qPCR验证结果表明,与正常组相比,6个基因都下调表达,其中MYLK、PKD1、MYH11、SOD3、TAGLN基因下调表达是有显著性差异的(P≤0.05),FLNA基因没有显著性差异(P>0.05)。结论:通过全基因组表达谱芯片可以筛选主动脉夹层与正常主动脉表达差异基因,同时结合RT-qPCR验证,为研究主动脉夹层提供新的思路。
OBJECTIVE: To study the gene expression profiling of human aortic dissection and normal aorta using gene chip technique. Methods: Five cases of aortic dissection and four cases of normal aorta were selected. Total RNA was extracted and reverse transcribed into cDNA. After in vitro transcription aRNA was hybridized with the chip, and the results were analyzed. At the same time, six differential genes including MYLK, PKD1, MYH11, SOD3, FLNA and TAGLN were screened for the quantitative verification of gene transcription level. RESULTS: A total of 1 661 genes were found in the human aortic dissection group compared with the normal aorta group, with 997 genes up-regulated and 664 genes down-regulated. The results of RT-qPCR analysis of 6 genes showed that 6 genes were down-regulated compared with normal group, of which MYLK, PKD1, MYH11, SOD3 and TAGLN genes were significantly down-regulated (P≤0.05) No significant difference (P> 0.05). Conclusion: Differential gene expression in aortic dissection and normal aorta can be screened by genome-wide cDNA microarray. Simultaneously RT-qPCR and RT-qPCR can provide a new idea for the study of aortic dissection.