骨形态发生蛋白2诱导成骨分化与长链非编码RNA AK007000的影响

Osteogenic differentiation induced by bone morphogenetic protein 2 and long non-coding RNA AK007000

背景:长链非编码RNA调控一系列生理过程,被认为在发育、分化和代谢的基因调控中发挥重要的作用。MC3T3-E1、C2C12和C3H10T1/2细胞可向骨细胞、肌细胞等多个方向分化,用于肌肉骨骼等运动系统相关疾病的研究。目的:观察长链非编码RNA在骨形态发生蛋白2诱导成骨分化中的作用。方法:对MC3T3-E1、C2C12和C3H10T1/2细胞在骨形态发生蛋白2诱导下,成骨分化过程中的长链非编码RNA表达变化进行芯片分析,找到在3株细胞中同时变化的长链非编码RNA,siRNA干扰方法观察长链非编码RNA对骨形态发生蛋白2诱导成骨分化的影响,采用Real-Time PCR与碱性磷酸酶染色检测成骨相关指标。结果与结论:骨形态发生蛋白2诱导MC3T3-E1、C2C12和C3H10T1/2成骨分化过程中,相应成骨指标增高,成肌指标肌细胞生成素降低。筛选出骨形态发生蛋白2诱导成骨分化过程中出发挥作用的长链非编码RNA AK007000。AK007000被干扰后成骨分化指标碱性磷酸酶、骨钙素、RUNX2、SP7表达下降,肌细胞生成素表达上升。因此,AK007000可能具有促进成骨抑制成肌作用。

BACKGROUND：Long non-coding RNA （lncRNA） regulates a series of physiological processes and it is considered to play important roles in the gene regulation of development, differentiation and metabolism. MC3T3-E1, C2C12 and C3H10T1/2 cells are able to differentiate into different celllineages, such as bone cells and muscle cells, and they can be used in the study of musculoskeletal diseases. OBJECTIVE：To study the role of lncRNA in osteogenic differentiation induced by bone morphogenetic protein 2. METHODS：Osteogenic differentiation of MC3T3-E1, C2C12 and C3H10T1/2 cells was induced by bone morphogenetic protein 2, and microarray expression profiling of lncRNA was undertaken in osteogenic differentiation. LncRNA simultaneous changes in three cells were found out. The siRNA interference of the lncRNA was used to study its effects on the osteogenic differentiation induced by bone morphogenetic protein 2. Real-time PCR and alkaline phosphatase staining were applied to detect osteogenesis related indicators. RESULTS AND CONCLUSION：In the process of osteogenic differentiation induced by bone morphogenetic protein 2, osteogenic differentiation indicators were increased, while myogenic differentiation indicator myogenin was reduced. LncRNA AK007000 was screened out to play a role in osteogenic differentiation induced by bone morphogenetic protein 2. Knockdown of lncRNA AK007000 decreased the expression of osteogenic differentiation indicators, while increased the expression of myogenin. Therefore, AK007000 may play a role in promoting osteogenic differentiation and inhibiting myogenic differentiation.