lncRNA靶向调控miR-133b在小型猪牙胚早期发育中的作用机制

非编码RNA在牙齿发育中发挥着重要的作用。我们前期在模型动物“小型猪”中证实，miR-133b是牙齿形态发生的关键miRNA，可靶向抗凋亡分子Mcl-1；还发现5个特异性表达的lncRNA与miR-133b表达趋势相反，并与之存在碱基互补，可能为竞争性内源RNA(ceRNA)。我们认为：某些“关键lncRNA”可发挥ceRNA活性，通过拮抗miR-133b调控Mcl-1的作用，进而影响凋亡，参与牙齿形态发生的调控。据此，本课题拟采用RNA pull-down、REMSA、RIP等方法，验证miR-133b通过Mcl-1影响牙胚细胞凋亡的信号通路，筛选对miR-133b有抑制性作用的“关键lncRNA”，探索Mcl-1是否进入细胞核及与“关键 lncRNA”的关系。力图揭示lncRNA/miR-133b/Mcl-1三者在牙胚早期发育中的相互作用机制，为基于发育模式的牙再生提供新的理论依据。

Non encoding RNAs play important roles in the tooth development. Our previous studies confirmed that miR-133b is the key miRNA during tooth morphogenesis by targeting anti-apoptotic molecule Mcl-1in the animal model of " miniature swine ". Five specific expression of LncRNAs were also found to have the opposite expression trends compared with miR-133b with the presence of nucleotide complementary which may act as the competition of endogenous RNA (ceRNA). We suggest that some of the "key lncRNAs" can act as ceRNA, participate in the regulation of tooth morphogenesis through targeting miR-133b, and affect apoptosis by regulation of Mcl-1. Accordingly, this study intends to use RNA pull-down, REMSA, RIP to validate key lncRNAs which suppress the miR-133b, further concentrate on the miR-133b/ Mcl-1 apoptosis mechanisms, and explore whether Mcl-1 can switch to the nucleus and interact with the key lncRNAs. Thus we can reveal the mechanisms of LncRNA/miR-133b/Mcl-1 during early development of tooth germ and provide a new theoretical basis for the regeneration of tooth based on development model.