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目的:分析高表达Aurora-A的细胞株KYSE150/GFP-Aur与对照细胞株KYSE150/GFP之间的基因表达谱差异,探讨Aurora-A高表达与相关基因表达差异之间的相关性。方法:按一步法抽提KYSE150/GFP-Aur与KYSE150/GFP细胞的总RNA,将其纯化后逆转录,以Cy5和Cy3标记cDNA作为探针,在表达谱芯片上进行杂交。经激光扫描仪扫描荧光信号图像,采用GenePix Pro4.0图像分析软件对芯片图像进行数字化处理和分析。结果:按差异显著性标准筛选出差异表达基因139个,其中呈上调表达的基因97个,呈下调表达的基因42个。Aurora-A高表达导致了多个肿瘤发生发展相关基因的表达变化,主要涉及细胞凋亡、增殖、侵袭、转移、DNA修复和细胞周期调控等5类基因。结论:对该表达谱芯片的分析能够高通量筛选一组Aurora-A高表达后发生表达改变的基因,为揭示肿瘤发生发展的分子机制提供了新的研究思路。
OBJECTIVE: To analyze the difference of gene expression profiles between KYSE150 / GFP-Aur cells and KYSE150 / GFP cells, and to explore the relationship between Aurora-A high expression and related gene expression differences. Methods: The total RNA of KYSE150 / GFP-Aur and KYSE150 / GFP cells was extracted by one-step method. After purification, the total RNA was reverse transcribed. Cy5 and Cy3-labeled cDNAs were used as probes to perform hybridization on the expression microarray. Fluorescent signal was scanned by laser scanner and digitized and analyzed by GenePix Pro4.0 image analysis software. RESULTS: A total of 139 differentially expressed genes were screened by the criteria of difference significance, among which 97 were up-regulated genes and 42 were down-regulated genes. Aurora-A overexpression leads to the expression of several genes involved in tumor development and progression, mainly involving five types of genes: apoptosis, proliferation, invasion, metastasis, DNA repair and cell cycle regulation. CONCLUSIONS: The analysis of the expression profiling chip can screen a group of high-throughput genes whose expression changes after high expression of Aurora-A, providing a new research idea for revealing the molecular mechanism of tumorigenesis.