cDNA基因芯片分析人心肌细胞能量代谢相关基因

Analysis of energy metabolism related genes in human myocardial cell by cDNA microarray

目的通过对正常成年人心肌细胞 mRNA 表达差异性的分析,探讨心肌细胞能量代谢相关基因的变化,为今后全方面的研究和比较心肌的能量代谢的基因特点及变化趋势提供对比参数、为实现较早的对心肌细胞功能状态进行评价。方法将人心肌细胞相关的3 368个 cDNA 制备成基因芯片,应用 PCR、测序、荧光探针标记、双重杂交等方法,研究正常成人心肌细胞 RNA 与正常成人多组织参考 RNA 比较的基因差异表达。结果对成人已知心肌细胞3 268个基因进行功能分组分析,得到高于成人多组织 RNA 对照1.5倍的基因576个,低于成人多组织 RNA 对照1.5倍的基因有1199个。与细胞能量代谢相关的基因有194个,其中有17个高表达,4个低表达;与细胞骨架和动力相关的基因有65个,其中有18个高表达,1个低表达。结论正常成人心肌细胞中存在许多相对于成人多组织 RNA 的特异标识基因,可为研究心血管疾病不同阶段不同病理状态提供客观参考体系。

Objective We selected 3 268 genes of human myocardial cells and analyzed the variability of mRNA expression of myocardial cells in normal adults, to explore the variation of energy metabolism related genes in human myocardial cell and compared the characteristics and variation tendency of energy metabolism related genes in human myocardial cells in the early stage. Methods Microarray was prepared with 3 368 myocardial cell related cDNAs. And we studied differentially expressed genes between myocardial cells and multi-tissues in normal adults by PCR, sequence, fluorescent probe labelling and double hybridisation. Results Functional group analysis was carried out with 3 268 genes of myocardial cells in normal adults. Compared with Universal Human Reference, 576 genes from myocardial cells were upregulated （≥ 1.5 folds）. Expression of 1 199 genes were down-regulated（ ≤66. 7% ）. 194 genes were involved in cell energy metabolism. Among them, 17 genes were up-regulated and 4 genes were downregulated. 65 genes were related with cellular skeleton, 18 of them had up-regulation and one of them had down-regulation. Conclusion Compared with multi-tissues, there were many genes preferentially expressed in myocardial cells of normal adults, which can be used as a reference system to evaluate the different stages and different pathological behaviors of cardiovascular diseases.