摘要
目的探讨正常与激素干预的大鼠成骨细胞的差异表达miRNA,并预测其靶基因。方法取SD乳鼠的成骨细胞进行培养、传代,取第3代成骨细胞分为空白组和激素组,空白组用含10%大鼠血清的L-DMEM培养基培养,激素组用含10^(-5)mol/L甲泼尼龙、10%大鼠血清的L-DMEM培养基;分别提取总RNA并进行质检;采用RNA标记与芯片杂交,采集数据及标准化,找出两组间的差异表达miRNA并对其表达谱进行分析。利用miranda、microcosm、mirdb 3个靶点预测软件对最明显的miRNA进行靶基因预测。结果与空白组相比,激素组中倍数>2.0(表达上调)的miRNA有4个,倍数<0.5(表达下调)的miRNA有2个,其中以上调的miR-672-5p和下调的miR-146a-5p最明显,其分别具有4个和11个可能靶基因。结论激素干预后大鼠成骨细胞的miRNA表达发生了明显变化,以miR-672-5p和miR-146a-5p最为显著。这表明它们有可能参与成骨细胞的凋亡和股骨头坏死的发生。
Objective To investigate the differential expression of miRNA between normal osteoblasts and those after hormone intervention in rats, and to predict their target RNA. Methods The osteoblasts of neonatal SD rats were cultured and passaged. The third generation osteoblasts were divided into the blank group and hormone group. The blank group was cultured in L - DMEM medium with 10% rats serum, while the hormone group was cultured in L - DMEM me- dium with 10-5 mol/L methylprednisolone and 10% rats serum. The total RNA was extracted and the quality control was performed. The data were collected and standardized by RNA marker and chip hybridization to identify differentially expressed mRNA, whose expression profile was analyzed. Miranda, microcom and mirdb target point prediction softwares were used for the most obvious miRNA target gene prediction. Results Compared with the blank group, 4 miRNAs were up - regulated （ ratio 〉 2. 0） and 2 miRNAs were down - regulated （ ratio 〈 0. 5 ） in the hormone group, the most obvious being up -regulation of miR -672 -5p and down -regulation of miR - 146a -5p. The numbers of predicting target genes in miR -672 -5p and miR - 146a -5p were 4 and 11, respectively. Conclusion The expression of miRNA in osteo- blasts of rats after hormone intervention is significantly changed, and miR - 672 - 5p and miR - 146a - 5p are the most evident. This suggests that it may be involved in the apoptosis of osteoblasts and occurrence of steroid induced osteonecro- sis of the femoral head.
出处
《广东医学》
CAS
北大核心
2016年第9期1261-1264,共4页
Guangdong Medical Journal
基金
国家自然科学基金资助项目(编号:81373652)
