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目的研究转染叉头蛋白O1(FOXO1)后A549细胞基因表达谱的改变,为深入探讨FOXO1在急性肺损伤中的作用机制提供线索。方法应用肿瘤坏死因子α(TNF-α)刺激A549细胞,应用脂质体介导的转染方法将真核表达载体GV230-FOXO1转染入经TNF-α诱导的A549细胞,再提取细胞的总RNA,应用基因芯片技术进行分析。结果成功构建并验证了FOXO1真核表达载体,提取了高质量的mRNA进行芯片分析,并通过RNA质量控制(RNA QC)验证。芯片分析结果显示差异表达基因中上调的基因有317个,下调的基因有237个。这些差异表达的基因功能涉及细胞增殖、凋亡、分化等多个方面。结论应用基因芯片技术可成功筛选转染FOXO1基因后A549细胞的差异表达基因。FOXO1在急性肺损伤中可能通过改变细胞的增殖、凋亡与分化等水平来影响疾病的进程。
Objective To investigate the gene expression profiles of A549 cells transfected with forkhead O1 (FOXO1), and to provide clues for further study on the mechanism of FOXO1 in acute lung injury. Methods A549 cells were stimulated with tumor necrosis factor-α (TNF-α) and transfected into A549 cells induced by TNF-α by liposome-mediated transfection method. RNA, using gene chip technology for analysis. Results The FOXO1 eukaryotic expression vector was successfully constructed and verified. High-quality mRNA was extracted for chip analysis and verified by RNA quality control (RNA QC). Chips analysis showed that there were 317 up-regulated genes among the differentially expressed genes and 237 down-regulated genes. These differentially expressed genes involved in many aspects of cell proliferation, apoptosis, differentiation and so on. Conclusion The differentially expressed genes of A549 cells transfected with FOXO1 gene can be successfully screened by gene chip technology. FOXO1 may affect the course of the disease in acute lung injury by altering cell proliferation, apoptosis and differentiation.