Li Lina, Fang Jie, Liu Yi, Xiao Li
Geriatric & VIP Department, Sichuan Academy of Medical Sciences & Sichuan Provincial People's Hospital, Chengdu, China.
State Key Laboratory of Oral Diseases, National Clinical Research Center for Oral Diseases, West China Hospital of Stomatology, Sichuan University, Chengdu, China.
PeerJ. 2020 Apr 15;8:e8909. doi: 10.7717/peerj.8909. eCollection 2020.
Osteogenic differentiation is an important role in dental implantation. Long no coding RNAs (lncRNAs) are a novel class of noncoding RNAs that have significant effects in a variety of diseases. However, the function and mechanisms of LOC100506178 in osteogenic differentiation and migration of bone morphogenetic protein 2 (BMP2)-induced osteogenic differentiation of human bone marrow mesenchymalstem cells (hBMSCs) remain largely unclear. BMP2 was used to induce osteogenic differentiation of hBMSCs. Quantitative real time PCR (qRT-PCR) was used to examine the expression of LOC100506178, miR-214-5p, Runt-related transcription factor 2 (RUNX2), Osterix (Osx), and Alkaline Phosphatase (ALP) in BMP2-induced osteogenic differentiation of hBMSCs. The function of LOC100506178 and miR-214-5p was explored in vitro using Alizarin Red S Staining, ALP activity, as well as in vivo ectopic bone formation. Luciferase reporter assay was performed to assess the association between LOC100506178 and miR-214-5p, as well as miR-214-5p and BMP2. The miR-214-5p sponging potential of LOC100506178 was evaluated by RNA immunoprecipitation. In the present study, the expression of LOC100506178 was found to be increased in BMP2-induced osteogenic differentiation of hBMSCs, accompanied with decreased miR-214-5p expression and increased RUNX2, Osx and ALP expression. LOC100506178 significantly induced, while miR-214-5p suppressed the BMP2-induced osteogenic differentiation of hBMSCs. Mechanistically, LOC100506178 was directly bound to miR-214-5p and miR-214-5p targeted the 3'-untranslated region of BMP2 to negatively regulate its expression. In conclusion, our data indicate a novel molecular pathway LOC100506178/miR-214-5p/BMP2 in relation to hBMSCs differentiation into osteoblasts, which may facilitate bone anabolism.
成骨分化在牙种植中起着重要作用。长链非编码RNA（lncRNAs）是一类新型的非编码RNA，在多种疾病中具有显著作用。然而，LOC100506178在骨形态发生蛋白2（BMP2）诱导的人骨髓间充质干细胞（hBMSCs）成骨分化和成骨细胞迁移中的功能及机制仍不清楚。采用BMP2诱导hBMSCs成骨分化。运用定量实时PCR（qRT-PCR）检测LOC100506178、miR-214-5p、 runt相关转录因子2（RUNX2）、osterix（Osx）和碱性磷酸酶（ALP）在BMP2诱导的hBMSCs成骨分化中的表达。通过茜素红S染色、ALP活性体外实验以及体内异位骨形成实验探究LOC100506178和miR-214-5p的功能。进行荧光素酶报告基因实验以评估LOC100506178与miR-214-5p以及miR-214-5p与BMP2之间的关系。通过RNA免疫沉淀评估LOC100506178的miR-214-5p海绵化潜力。在本研究中，发现LOC100506178在BMP2诱导的hBMSCs成骨分化中表达增加，同时miR-214-5p表达降低，RUNX2、Osx和ALP表达增加。LOC100506178显著诱导，而miR-214-5p抑制BMP2诱导的hBMSCs成骨分化。机制上，LOC100506178直接与miR-214-5p结合，miR-214-5p靶向BMP2的3'-非翻译区以负向调节其表达。总之，我们的数据表明了一条与hBMSCs向成骨细胞分化相关的新型分子途径LOC100506178/miR-214-5p/BMP2，这可能促进骨合成代谢。
