长链非编码RNA- NR_028308及其靶基因在先天性小耳畸形中调控机制的研究

Detection of long chain non encoding RNA NR_028308 with a normal side auricular cartilage and in patients with congenital microtia side expression of auricular cartilage in the comparison of paired difference expression between tissue, cell lines, through the isolation and culture of normal and abnormal small ear cartilage tissue and biological characteristics analysis, LncRNA-NR_028308 and TNFRSF11B etc. the target gene for further study of several aspects of the development and differentiation of cartilage cells in the study on the relativity of biological pathways. Objective to investigate the clinical significance of the expression of NR_028308 in the diagnosis of congenital microtia.

检测长链非编码核糖核酸NR_028308在同一位患者的正常侧耳廓软骨和先天性小耳畸形侧耳廓软骨中的表达水平，比较配对组织间、细胞系之间的表达差异，通过正常和畸形小耳软骨组织的分离培养及生物学特性分析，LncRNA-NR_028308及TNFRSF11B等靶基因对软骨细胞生物学通路中软骨发育分化的相关性研究等几方面深入研究。分析表达上调的NR_028308对先天性小耳畸形的表观遗传学影响，探讨其在小耳畸形中的临床意义。

本课题组通过验证LncRNA-ENST00000449766及ZFP36L2，LncRNA NR_028308与OPG在残耳软骨中较正常耳软骨中高表达，并且敲低lncRNA后，其靶基因表达相应下调，敲低lncRNA及靶基因后，软骨外基质中的蛋白成分相应升高，软骨细胞增殖得到促进,证明LncRNA NR_028308、LncRNA-ENST00000449766及其靶基因可抑制残耳细胞分泌软骨外基质以及抑制软骨细胞增殖。我们通过进行转录组芯片检测，发现残耳软骨及正常软骨间的差异分子主要与细胞外基质合成相关，通过构建ceRNA网络，得到lncRNA与靶基因间的桥梁miRNA,通过原位免疫杂交明确lncRNA与miRNA的共定位，通过RIP-seq得到靶基因ZFP36L2可结合的RNA,结合转录组结果分析ZFP36L2基因的下游基因SEC13,明确了ZFP36L2影响残耳细胞增殖及细胞外基质合成的分子机制。