Comparative and quantitative proteomic analysis of normal and degenerated human annulus fibrosus cells

Abstract

Summary Degeneration of the intervertebral disc (IVD) is a major chronic medical condition associated with back pain. To better understand the pathogenesis of IVD degeneration, we performed comparative and quantitative proteomic analyses of normal and degenerated human annulus fibrosus (AF) cells and identified proteins that are differentially expressed between them. Annulus fibrosus cells were isolated and cultured from patients with lumbar disc herniation (the experimental group, degenerated AF cells) and scoliosis patients who underwent orthopaedic surgery (the control group, normal AF cells). Comparative proteomic analyses of normal and degenerated cultured AF cells were carried out using 2‐D electrophoresis, mass spectrometric analyses, and database searching. Quantitative analyses of silver‐stained 2‐D electrophoresis gels of normal and degenerated cultured AF cells identified 10 protein spots that showed the most altered differential expression levels between the two groups. Among these, three proteins were decreased, including heat shock cognate 71‐kDa protein, glucose‐6‐phosphate 1‐dehydrogenase, and protocadherin‐23, whereas seven proteins were increased, including guanine nucleotide‐binding protein G(i) subunit α‐2, superoxide dismutase, transmembrane protein 51, adenosine receptor A3, 26S protease regulatory subunit 8, lipid phosphate phosphatase‐related protein, and fatty acyl‐crotonic acid reductase 1. These differentially expressed proteins might be involved in the pathophysiological process of IVD degeneration and have potential values as biomarkers of the degeneration of IVD.