基因芯片分析鱼藤素对斑马鱼胚胎发育的影响

Deguelin treatment delays embryonic development of zebrafish: A gene chip analysis

背景:鱼藤素具有延缓斑马鱼胚胎发育的作用,但其具体作用机制尚未明确。目的:分析鱼藤素处理斑马鱼胚胎的基因与信号传导通路表达差异。方法:斑马鱼胚胎来自于根据Zebrafish book中描述的方法养的殖斑马鱼。用0.6μmol/L鱼藤素处理2-细胞期斑马鱼胚胎,以二甲基亚砜孵化液处理2-细胞期斑马鱼胚胎为对照。4 hpf时期提取二甲基亚砜组及鱼藤素组总RNA,利用基因芯片技术检测鱼藤素处理斑马鱼胚胎后基因的差异表达,采用实时荧光定量PCR验证基因芯片结果,结合聚类分析及通路分析来探索鱼藤素的作用机制。结果与结论:芯片结果显示,鱼藤素处理斑马鱼胚胎后差异表达上调3倍以上的基因407个,下调的461个。PCR验证结果与芯片结果相符。经KEGG通路分析,涉及14条通路。鱼藤素可能通过干扰细胞的代谢生长及影响其分化起作用。

BACKGROUND： Anticancer drug deguelin delays the embryonic development of zebrafish, but its exact mechanism is not yet clear. OBJECTIVE： To explore expression differences in genes and signaling pathways in deguelin-treated zebrafish embryos. METHODS： Zebrafish embryos were harvested from zebrafish fed according to the Zebrafish Book. Zebraflsh embryos at 2-cell stage were selected and divided into two groups： control group treated with dimethyl sulfoxide incubator liquid, and experimental group treated with 0.6 pmol/L deguelin. Total RNA was extracted from the two groups at sphere stage. Then, gene chip technique was used to detect differentially expressed genes in the deguelin-treated zebraflsh embryos. Real-time quantitative PCR was employed to validate microarray cluster analysis and pathway analysis to explore the mechanism of action of deguelin. RESULTS AND CONCLUSION： Chip results showed that 407 genes were upregulated more than three times and 461 genes were downregulated more than three times after deguelin treatment. PCR validation results were consistent with those of the chip. Fourteen pathways were identified by KEGG pathway analysis. Deguelin may play an important role by intervening cell metabolism growth and differentiation.