Identification of suitable reference gene and biomarkers of serum miRNAs for osteoporosis.
Our objective was to identify suitable reference genes in serum miRNA for normalization and screen potential new biomarkers for osteoporosis diagnosis by a systematic study. Two types of osteoporosis models were used like as mechanical unloading and estrogen deficiency. Through a large-scale screening using microarray, qPCR validation and statistical algorithms, we first identified miR-25-3p as a suitable reference gene for both type of osteoporosis, which also showed stability during the differentiation processes of osteoblast and osteoclast. Then 15 serum miRNAs with differential expression in OVX rats were identified by microarray and qPCR validation. We further detected these 15 miRNAs in postmenopausal women and bedrest rhesus monkeys and evaluated their diagnostic value by ROC analysis. Among these miRNAs, miR-30b-5p was significantly down-regulated in postmenopausal women with osteopenia or osteoporosis; miR-103-3p, miR-142-3p, miR-328-3p were only significantly decreased in osteoporosis. They all showed positive correlations with BMD. Except miR328-3p, the other three miRNAs were also declined in the rhesus monkeys after long-duration bedrest. Their AUC values (all >0.75) proved the diagnostic potential. Our results provided a reliable normalization reference gene and verified a group of circulating miRNAs as non-invasive biomarkers in the detection of postmenopausal- and mechanical unloading- osteoporosis.
Institute of Cell and Development Biology, College of Life Sciences, Zhejiang University, Hangzhou, 3100058, China.|State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Jian
J
Chen
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Kai
K
Li
Department of Endocrinology, Peking Union Medical College Hospital, Peking Union Medical College, Beijing, 100032, China.
Qianqian
Q
Pang
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Chao
C
Yang
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Hongyu
H
Zhang
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Feng
F
Wu
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Hongqing
H
Cao
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Hongju
H
Liu
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Yumin
Y
Wan
Department of Endocrinology, Peking Union Medical College Hospital, Peking Union Medical College, Beijing, 100032, China.
Weibo
W
Xia
Institute of Cell and Development Biology, College of Life Sciences, Zhejiang University, Hangzhou, 3100058, China.
Jinfu
J
Wang
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Zhongquan
Z
Dai
State Key Laboratory of Space Medicine Fundamentals and Application, China Astronaut Research and Training Center, Beijing, 100094, China.
Yinghui
Y
Li
D000818:Animals; D015415:Biomarkers / Q000097:blood*; D004195:Disease Models, Animal; D005260:Female; D005786:Gene Expression Regulation; D006801:Humans; D008253:Macaca mulatta; D035683:MicroRNAs / Q000097:blood*; D020411:Oligonucleotide Array Sequence Analysis / Q000592:standards*; D010024:Osteoporosis / Q000097:blood / Q000235:genetics*; D017698:Postmenopause; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*
D015415:Biomarkers; C543351:MIRN25 microRNA, human; D035683:MicroRNAs