肺栓塞-深静脉血栓形成相关基因群表达谱变化的研究

An study on screening the gene clusters associated with pulmonary embolism-deep venous thrombosis by oligo microarray

目的研究肺栓塞患者与正常对照者基因表达谱的差异,探讨肺栓塞发生的分子机制。方法收集9例肺栓塞患者和33例正常对照的新鲜血样,分离血总 RNA。合成9个 Cy3标记的肺栓塞患者 cRNA 探针和1个 Cy5标记的标准对照探针,与44K 人全基因组(Oligo)芯片杂交,且行实时荧光 PCR 验证。结果 9张芯片筛选出共同差异表达基因434条,涉及血液凝固36条、免疫炎症20条、代谢29条、分化凋亡26条、细胞生长维持25条、细胞信号传递22条、细胞骨架和运动14条、离子通道和运输15条、转录14条、RNA/DNA 结合6条及其他。随机选取其中11条基因进行FQ-PCR 扩增,结果与芯片相符。结论肺栓塞-深静脉血栓形成的发生和演变过程中,体内有多种基因出现了差异性表达。

Objective To find out the gene clusters associated with pulmonary embolism-deep venous thrombosis（ PE-DVT） and elucidate the molecular genetic mechanism of PE. Methods Peripheral venous blood samples were collected from 9 PE patients and 33 normal controls. The total RNA was extracted and purified. Nine PE cRNA probes labeled with cyanine 3 were constructed, and one standard cRNA probe labeled with cyanine 5 was synthesized based on the total RNA mixture of the controls. Hybridization with Agilent Whole Human Genome Oligo Microarray was performed. Eleven of the genes screened were randomly selected to be amplified by fluorescence quantitative PCR （FQ-PCR）. Results 434 differential expression genes were screened from the 9 microarrays, including 36 gene transcripts. Associated with blood coagulation, 20 with immune or inflammatory response, 29 with metabolism, 26 with cell differentiation and apoptosis, 25 with cell growth/maintenance, 22 with cell-cell signaling or signal transduction, 14 with cytoskeleton or motility, 15 with ion channel or ion transport, 14 with transcription, and 6 with DNA/RNA binding. These 11 of these genes were randomly selected to be amplified by FQ-PCR. The differential expression of the 11 selected genes was consistent with the microarray. Conclusion The differential expression of a lot of genes in the body may play a role in the process of initiation and development of PE- DVT.