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目的筛选BMP-2诱导成骨分化过程中表达上调LncRNAs,并进行功能研究。方法用BMP-2分别诱导MC3T3-E1、C2C12和C3H10T1/2细胞进行成骨分化,并利用芯片技术分析这一过程中差异表达LncRNAs与mRNA,找到在三株细胞中同时上调1.5倍的LncRNAs,应用基因本体项目分析和Fisher精确测试预测LncRNAs调控的靶基因。RT-qPCR验证芯片结果,采用RNAi方法下调相应LncRNAs表达水平后分析其与BMP-2诱导成骨分化的关系。结果筛选出在三株细胞中同时上调1.5倍增LncRNAs 39个,RT-qPCR验证结果与芯片检测结果符合率为50.9%。干扰AK051397、AK039899后成骨分化标记基因ALP、OC、SP7表达明显下降(P<0.05)。结论 AK051397与AK039899是BMP-2诱导成骨分化过程中的正向调节因子。
OBJECTIVE: To screen and up-regulate LncRNAs induced by BMP-2 during osteogenic differentiation and to study their function. Methods The osteogenic differentiation of MC3T3-E1, C2C12 and C3H10T1 / 2 cells was induced by BMP-2, and LncRNAs and mRNA were differentially expressed in this process by using chip technology. LncRNAs that were simultaneously up-regulated by 1.5 times in three cells were found, Prediction of target genes regulated by LncRNAs using gene ontology project analysis and Fisher exact test. RT-qPCR verification of the results of the chip, RNAi method down-regulated LncRNAs expression levels were analyzed BMP-2 induced osteogenic differentiation relationship. RESULTS: Thirty-nine LncRNAs were screened at the same time in 39 cells, 39 of them were screened out. The coincidence rate of RT-qPCR and chip test was 50.9%. After AK051397 and AK039899 were interfered, the expressions of ALP, OC and SP7 gene were significantly decreased (P <0.05). Conclusion AK051397 and AK039899 are positive regulators of osteogenic differentiation induced by BMP-2.