miR-3175在翼状胬肉上皮-间质转化中的作用及机制研究

Pterygium is a common degenerative disease of ocular surface, which have high postoperative reoccurrence rate, however, the mechanism of it is still unclear. Recent studies indicated that the molecular mechanism of pterygium might be related with EMT (epithelial-mesenchymal transition), which was validated by our previous research. Furthermore, we found that miR-3175 is an important regulator among the EMT process in pterygium, which could be regulated by TGF- beta/Smad signaling pathway. In this study, we focus on miR-3175, in an effort to understand its function in EMT and pterygium pathogenesis. First, we transfect pterygial epithelium cells with mir-3175 expression vectors (over-expression vectors and inhibitors). Second, we investigate the potential targeted genes of miR-3175 through CLIP sequencing (cross-linking and immunoprecipitation sequencing). Finally, we will elucidate the function of miR-3175 and build a molecular regulating network of miR-3175 in EMT of pterygial epithelium by bioinformatics and molecular biology technology. The study will help us to provide a new molecular mechanism of pterygium pathogenesis, which might improve the incidence and recurrence among pterygium patients.

翼状胬肉是眼科常见的眼表疾病，然而其发病机制目前仍不明确，手术治疗也存在较高的复发率。最新的研究认为翼状胬肉的分子病理学机制可能与上皮-间质转化（EMT）有关。本项目前期研究发现，翼状胬肉上皮与正常结膜上皮相比，确实发生了显著的EMT现象，并发现miR-3175可能通过TGF-beta/Samd信号通路诱导翼状胬肉上皮细胞EMT。因此，本研究拟通过构建miR-3175表达载体，转染体外培养的翼状胬肉上皮细胞，采用交联免疫沉淀技术解析miR-3175直接作用的靶基因，利用生物信息学及分子生物学手段研究miR-3175在翼状胬肉EMT中的靶基因及其调控路径，构建miR-3175在翼状胬肉上皮EMT中的调控网络，明确其在翼状胬肉上皮EMT中的生物学功能。以期进一步揭示翼状胬肉形成的分子机制，为阻止翼状胬肉的发生和降低翼状胬肉的术后复发率提供新的作用靶点和干预策略。

翼状胬肉是眼科常见的眼表疾病，然而其发病机制目前仍不明确，手术治疗也存在较高的复发率。最新的研究认为翼状胬肉的分子病理学机制可能与上皮-间质转化有关。本项目前期研究发现，翼状胬肉上皮与正常结膜上皮相比，确实发生了显著的EMT现象。因此，本研究通过体外构建miR-3175表达载体，转染表型与翼状胬肉上皮细胞一致的CCL细胞，发现miR-3175上调可显著促进CCL细胞EMT的发生，并采用交联免疫沉淀技术解析miR-3175直接作用的靶基因，利用生物信息学及分子生物学手段确定miR-3175在CCL细胞EMT中直接作用的的靶基因Trim62，并验证了两者的关系，构建其细胞信号调控网络，明确其在翼状胬肉上皮EMT中的生物学功能。本研究进一步揭示翼状胬肉形成的分子机制，为阻止翼状胬肉的发生和降低翼状胬肉的术后复发率提供新的作用靶点和干预策略。