Discovery and identification of serum biomarkers for postmenopausal osteoporosis based on TMT labeling and HPLC-MS/MS technology

作者:Shi, Xiaolin; Liang, Bocheng; Shi, Zhenyu; Wang, Bo; Wu, Peng; Kong, Lingcheng; Yao, Jianliang; Li, Chunwen*
来源:International Journal of Clinical and Experimental Medicine, 2017, 10(1): 334-346.

摘要

Objective: To screen the serum protein molecular markers of postmenopausal osteoporosis by the proteomics analysis using Tandem Mass Tag (TMT) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Methods: Serum protein samples were recruited from 10 cases of postmenopausal patients with osteoporosis and 10 cases of postmenopausal women without osteoporosis and the high abundance ratios protein was removed, differentiation protein was extracted and labeled with TMT reagent. Then, mass spectrometric detection, data analysis of differentially expressed proteins, and analysis of biological information were carried out. Results: 87 significantly differentially expressed proteins were screened from the differentiated protein expression profile by LC-ESI-MS/MS combined with TMT labeling, including 50 proteins up-regulated and 37 proteins down-regulated. Differentially expressed proteins were analyzed by GO annotation, these proteins are mainly involved in 15 kinds of biological processes, seven kinds of cellular component and six kinds of molecular function. RAB7A, TSP1, GAS6, SPP24 were screened as candidate proteins which were related to the mechanism of bone remodeling of osteoporosis. By STRING10.0 protein interaction network analysis tools, RAB7A, TSP1, GAS6 were located in the center of the interaction network. SPP24 was located at edge of the network, but it is directly related to the protein BMP2 of bone remodeling. Conclusion: These results provide that the proteomics analysis by using TMT combined with LC-ESI-MS/MS was a feasible method for screening the molecular biomarkers. It suggests that RAB7A, TSP1, GAS6 and SPP24 may be a useful biomarker which can be used in diagnosis and treatment of postmenopausal osteoporosis.