维甲酸诱导腭裂相关wnt和成纤维细胞生长因子配体表达的动态变化

摘要

Objective To screen the wnt and fibroblast growth factor(FGF) ligands involved in palatogenesis and cleft palate, and to study the dynamic expression of them in the different stages of palatal development and cleft palate formation. Methods Mouse model of retinoic acid (RA)-induced cleft palate was set up. At embryo day (ED) 14.5, the palatal tissues of RA-treated group and wild type were collected and prepared for gene-chip analysis.According to the gene-chip results, wnt3, wnt8a, fgf9 and fgf10 were selected and their expression level was detected at ED13.5-15.5 by using semi-quantitative reverse transcription-PCR (RT-PCR). Results 1)Gene-chip analysis showed that in RA-induced cleft palate group wnt8a and fgf9 were down-regulated, wnt3 and fgf10 were up-regulated in conversely. 2)During the different stage of the control group palatogenesis, intense expression of wnt3, wnt8a, fgf9 and fgf10 were detected with a continuous dynamic pattern. 3)Compared with the control group, the expression level of wnt3, wnt8a, fgf9 and fgf10 in RA-induced cleft palate showed significant difference, respectively (P＜0.05).Conclusion wnt and FGF signaling molecules participate in the palatogenesis, and RA pathway may interact with wnt and FGF signaling pathway.%目的 筛选与维甲酸诱导腭裂相关的wnt和成纤维细胞生长因子(FGF)信号分子,观察其在正常腭突发育和维甲酸诱导腭裂形成不同阶段的表达动态变化.方法建立维甲酸诱导腭裂小鼠模型,制作基因芯片并筛选wnt和FGF信号通路相关基因.根据芯片结果选出wnt经典通路配体wnt3和wnt8a、FGF通路配体fgf9和faf10,并应用半定量逆转录-PCR(RT-PCR)技术检测其在胚胎天数(ED)13.5～15.5中表达变化.结果 1)ED14.5实验组腭突组织芯片分析结果显示:wnt3和fgf10表达水平上调,wnt8a和fsf9表达水平下降.2)在ED13.5～15.5正常对照组腭突发育的不同阶段,上述基因呈现持续性高表达,表达水平呈动态变化.3)在ED13.5～15.5的实验组腭裂发生的不同阶段,上述基因的表达水平与正常对照组均有差异(P<0.05).结论 wnt和FGF信号分子通路参与小鼠胚胎继发腭的正常发育过程,在维甲酸诱导的腭裂发生过程中,存在维甲酸通路与wnt和FGF通路的相互作用.