长链非编码RNA MEF2C-AS1 调控 成骨细胞功能的机制研究

Study on mechanism of lncRNA MEF2C-AS1 regulating osteoblast function

摘要:

目的 探讨长链非编码RNA MEF2C-AS1 通过肌细胞增强因子2C（MEF2C ）基因对成骨细胞 的影响。方法 培养人成骨细胞系hFOB1.19，转染过表达MEF2C-AS1 质粒，检测过表达MEF2C-AS1 组 的碱性磷酸酶（ALP）活性变化；通过小干扰RNA 沉默MEF2C-AS1，用qRT-PCR、Western blotting 检 测MEF2C-AS1 水平变化对MEF2C 、SOST 基因的mRNA、蛋白水平的影响，再同时过表达MEF2C-AS1、 MEF2C，检测SOST 基因表达水平变化。结果 过表达MEF2C-AS1 组MEF2C、SOST mRNA 和蛋白相对 表达量较对照组低（P <0.05）。过表达MEF2C-AS1 组ALP 活性较对照组高（P <0.05）。沉默MEF2C-AS1 组MEF2C、SOST mRNA 和蛋白相对表达量较对照组高（P <0.05）。过表达MEF2C-AS1 组SOST mRNA 和蛋白相对表达量较对照组低（P <0.05）。过表达MEF2C-AS1+MEF2C 组SOST mRNA 和蛋白相对表达量 较过表达MEF2C-AS1 组高（P <0.05）。结论 MEF2C-AS1 可通过抑制MEF2C 进一步抑制SOST 基因的表 达，促进成骨细胞的分化成熟。

Abstract:

Objective To investigate whether long non-coding RNA (lncRNA) MEF2C-AS1 can affect the expression of SOST gene by acting on myocyte enhancer factors 2C (MEF2C) gene, thereby affecting the function of osteoblast. Methods Human osteoblast cell line hFOB1.19 was cultured and transfected with overexpressed MEF2C-AS1 plasmid, and the ALP activity of the overexpressed MEF2C-AS1 group was detected. By silencing MEF2C-AS1 with small interfering RNA, qRT-PCR and western blot experiments were conducted to detect the effects of MEF2C-AS1 level changes on the mRNA and protein levels of MEF2C and SOST genes. Then, the overexpression of MEF2C-AS1 and MEF2C was also conducted to detect the changes of SOST gene expression levels. Results Compared with control group, the mRNA levels of MEF2C and SOST in the overexpressed MEF2CAS1 group were decreased (P < 0.05), and the protein levels were also decreased (P < 0.05), but the ALP activity was increased (P < 0.05). The mRNA levels of MEF2C and SOST in the silenced MEF2C-AS1 group were increased (P < 0.05), and the protein levels were also increased (P < 0.05). The mRNA and protein expression levels of SOST gene in the over-expressed MEF2C and MEF2C-AS1 groups were both increased compared with that in the over-expressed MEF2C-AS1 group (P < 0.05). Conclusion This experiment confirmed that MEF2C-AS1 can inhibit the expression of SOST by inhibiting MEF2C, and promoting the differentiation and maturation of osteoblasts.