Journal Article

Down-regulated <i>HS6ST2</i> in osteoarthritis and Kashin–Beck disease inhibits cell viability and influences expression of the genes relevant to aggrecan metabolism of human chondrocytes

Wei Wang, Bo Zhong, Jian Sun, Junling Cao, Juan Tian, Nannan Zhong, Wenxiang Zhao, Lifang Tian, Peng Xu, Dawei Guo, Xichi Ju, Wei Ma, Meng Li, Weikun Hou and Shemin Lu

in Rheumatology

Volume 50, issue 12, pages 2176-2186
Published in print December 2011 | ISSN: 1462-0324
Published online October 2011 | e-ISSN: 1462-0332 | DOI: https://dx.doi.org/10.1093/rheumatology/ker230
Down-regulated HS6ST2 in osteoarthritis and Kashin–Beck disease inhibits cell viability and influences expression of the genes relevant to aggrecan metabolism of human chondrocytes

Show Summary Details

Preview

Objective. Primary OA and Kashin–Beck disease (KBD) show similar pathological changes in articular cartilage. The objective was to screen differentially expressed genes between OA and normal cartilage, confirm the candidate gene expression among OA, KBD and normal cartilage, and then clarify its role in vitro.

Methods. Differentially expressed genes in OA cartilage were screened by suppression subtractive hybridization (SSH) and verified by real-time quantitative PCR (Q-PCR) analysis. Heparan sulphate 6-O-sulphotransferase 2 (HS6ST2) expression was identified by Q-PCR and immunohistochemistry. After suppressing HS6ST2 by RNA interference in C28/I2 human chondrocyte line, the effects were analysed through determining the cell viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), the aggrecan contents by toluidine blue staining and the mRNA expression levels of SRY-type high mobility group box 9 (SOX9), AGC1, MMP3, a disintegrin and metalloproteinase domain with thrombospondin motifs 4 (ADAMTS4) and ADAMTS5 by Q-PCR.

Results. HS6ST2 in the reverse subtraction library was identified as a down-regulated gene in OA and KBD at both mRNA and protein levels. The percentage of safranion O staining area was correlated positively with the percentage of HS6ST2-positive chondrocytes in OA and KBD cartilage. After HS6ST2-specific short interfering RNA (siRNA) transfection to C28/I2 cells, the cell viability was inhibited significantly, and the mRNA expression levels of SOX9 and AGC1 were reduced markedly, while MMP3 expression was increased significantly.

Conclusion. HS6ST2 down-regulation was identified in both OA and KBD cartilage. The findings first suggest that HS6ST2 may participate in the pathogenesis of OA and KBD by influencing aggrecan metabolism.

Keywords: Osteoarthritis; Kashin–Beck disease; Suppression subtractive hybridization; Heparan sulphate 6-O-sulphotransferase 2

Journal Article.  5169 words.  Illustrated.

Subjects: Rheumatology

Full text: subscription required

How to subscribe Recommend to my Librarian

Users without a subscription are not able to see the full content. Please, subscribe or login to access all content. subscribe or purchase to access all content.