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为了探讨应用DNA微阵列研究骨髓增生异常综合征 (MDS)的基因表达谱的可行性 ,将 2例患者骨髓单个核细胞RNA各取等量混合后进行逆转录Cy5标记 ,与Cy3标记的正常对照cDNA混合 ,与H14 1微阵列杂交、扫描、软件分析。结果H14 1s芯片中共点样 13484个基因克隆 ,其中 10 6 4个靶克隆是由针对同一基因内不同序列的cDNA片段重复点样至少 2次。重复点样检测结果表明 :在 2张芯片内各自完全一致的分别为 6 2 5个 (5 8.7% )和6 30个 (5 9.2 % ) ,而存在完全相反结果的cDNA片段分别有 2 1个 (2 .0 % )和 11个 (1.0 % )。 10 6 4个靶克隆中重复点样数据完整且分别在 2张芯片内结果一致的共有 4 11个 ,其中在 2张芯片间结果也完全一致的共有 4 0 0个(97 3% )。 2张芯片中MDS与正常对照比较存在差异表达的靶基因分别为 15 4 9和 1311个 ,而 2张芯片间表达差异一致的靶基因为 4 0 9个 ,其中 10 1个基因参与造血调控 ,主要涉及转录因子、细胞周期调节蛋白、代谢相关基因、表面黏附分子等。结论 :DNA微阵列可用于对MDS患者混合标本的表达谱分析 ,为深入研究MDS的发病分子机理提供线索 ,但需要进行重复实验以降低微阵列操作过程引起的表达偏差。
In order to investigate the feasibility of using DNA microarray to study the gene expression profiles of myelodysplastic syndrome (MDS), 2 patients were treated with the same amount of RNA mixed with Cy5 and reverse transcribed with Cy3, cDNA hybridization, hybridization with H14 1 microarray, scanning, software analysis. Results A total of 13484 gene clones were spotted in the H14 1s chip. Among them, 106 target clones were spotted at least twice with cDNA fragments targeting different sequences in the same gene. The results of repeated spot detection showed that there were 6 2 5 (57.7%) and 6 30 (5 9.2%) respectively in the two chips, while the exact cDNA fragments with the opposite result were 21 (2.0%) and 11 (1.0%). A total of 4 10 target repeats with 10 4 4 target clones were found, with 4 1 1 (97 3%) identical results across 2 chips. There were 15 4 9 and 13 11 target genes differentially expressed in two of the MDSs compared with the normal controls, while the number of target genes with the same expression difference between the two chips was 409, of which 10 1 genes were involved in hematopoietic regulation, Mainly involved in transcription factors, cell cycle regulatory proteins, metabolism-related genes, surface adhesion molecules. Conclusion: DNA microarray can be used to analyze the expression profile of mixed samples in patients with MDS, providing clues for further study on molecular pathogenesis of MDS. However, repeated experiments are needed to reduce the expression deviation caused by microarray operation.