miR-30a-5p通过靶向Runx2基因抑制成骨细胞分化  

miR-30a-5p Inhibits Osteogenic Differentiation via Targeting Runx2 Gene

作　　者：王如然[1] 黄胜男[2] 曹戬 许华颖[1] 李宏燕[1] 黄浩然 赵杉[1] 王玉红 冯艳华[4] 徐国营[1] 韩兵[1] WANG Ru-Ran;HUANG Sheng-Nan;CAO Jian;XU Hua-Ying;LI Hong-Yan;HUANG Hao-Ran;ZHAO Shan;WANG Yu-Hong;FENG Yan-Hua;XU Guo-Ying;HAN Bing(Surgery Department,Guang'anmen Hospital,China Academy of Chinese Medical Science,Beijing 102618;Pharmacy Department,Guang'anmen Hospital,China Academy of Chinese Medical Science,Beijing 102618;Central Lab,Guang'anmen Hospital,China Academy of Chinese Medical Science,Beijing 102618;Hospital Infection-Control Department,Guang'anmen Hospital,China Academy of Chinese Medical Science,Beijing 102618;Surgery Department,Xi⁃hongmen Hospital,Daxing District,Beijing 100076,China)

机构地区：[1]中国中医科学院广安门医院南区外科,北京102618 [2]中国中医科学院广安门医院南区药剂科,北京102618 [3]中国中医科学院广安门医院南区中心实验室,北京102618 [4]中国中医科学院广安门医院南区院感科,北京102618 [5]北京市大兴区西红门医院外科,北京100076

出　　处：《生物技术通讯》2020年第2期142-147,共6页Letters in Biotechnology

基　　金：北京市中医药科技发展资金(JJ2018-59)。

摘　　要：目的:构建绝经后骨质疏松大鼠模型,分析miR-30a-5p在成骨细胞分化中的功能。方法:将SD大鼠分为雌激素组、假手术组和模型组;取胫骨组织进行miRNA测序并差异表达分析;检测miR-30a-5p在各组大鼠骨中的表达含量;通过qRT-PCR检测miR-30a-5p表达对成骨分化标记相关蛋白胶原Ⅰ、骨钙素(OCN)和骨桥蛋白(OPN)表达的影响,同时检测对碱性磷酸酶(ALP)活性的影响;双萤光素酶报告基因检测miR-30a-5p对RUNT相关转录因子2(Runx2)的靶向调控作用。结果:miRNA测序分析结果显示,相对于假手术组、雌激素组,模型组有33种共同miRNA表达上调,选取miR-30a-5p作为本研究对象;qRT-PCR进一步验证miR-30a-5p在模型组骨组织中高表达;miR-30a-5p靶向抑制Runx2的表达,且能够抑制OCN、OPN和胶原Ⅰ的表达,降低ALP活性;而同时过表达Runx2后,OCN、OPN和胶原Ⅰ的表达升高,ALP活性增加,Runx2过表达部分缓解miR-30a-5p对成骨细胞分化的抑制作用。结论:miR-30a-5p在骨质疏松大鼠骨组织中表达上调,并能够靶向抑制Runx2的表达,从而抑制成骨细胞分化,促进骨质疏松进展,为骨质疏松症的诊断提供新的生物标志物。Objective:To establish a postmenopausal osteoporosis rat model and explore the function of miR-30 a-5 p in osteoblast differentiation.Methods:SD rats were separated into estrogen group,sham-operated group and osteoporosis rat model group.miRNA sequencing was performed on rats tibia tissue,and the data analysis on differential expression in these three groups was carried out,and the expression of miR-30 a-5 p in rats tibia tissues was investigated by qRT-PCR.The effects of miR-30 a-5 p on the expression of osteoblast differentiation-associated markers,collagenⅠ,osteocalcin(OCN)and osteopontin(OPN)were detected.Dual luciferase gene reporter assay was used to determine whether RUNT-related transcription factor 2(Runx2)was the target gene of miR-30 a-5 p.Results:Venn diagram showed that the model group had 33 miRNAs that were up-regulated compared with the sham-operated group and the estrogen group.miR-30 a-5 p was chosen as the research object in this study.The high expression of miR-30 a-5 p in the tibia tissues of the model group was further verified by qRT-PCR.miR-30 a-5 p down-regulated the expression Runx2 via binding its 3’UTR,and inhibited the expression of collagenⅠ,OCN and OPN,as well as decreased the activity of alkaline phosphatase(ALP),which can be partially rescued by Runx2 overexpression.Conclusion:miR-30 a-5 p is up-regulated in the tibia tissues of osteoporosis rats and regulate the expression of Runx2.Consequently,miR-30 a-5 p promotes the progression of osteoporosis by inhibiting osteogenic differentiation via targeting Runx2,which provides a new biomarker for the diagnosis of osteoporosis.

关 键 词：miR-30a-5p RUNT相关转录因子2(Runx2) 成骨分化 骨质疏松 

分 类 号：Q78[生物学—分子生物学]