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
| Autor: | Meng Li, Junling Cao, Lifang Tian, Wenxiang Zhao, Dawei Guo, Weikun Hou, Xichi Ju, Juan Tian, Jian Sun, Peng Xu, Wei Wang, Nannan Zhong, Shemin Lu, Wei Ma, Bo Zhong |
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| Rok vydání: | 2011 |
| Předmět: |
Adult
Cartilage Articular Male Small interfering RNA DNA Complementary Cell Survival Down-Regulation Gene Expression Biology Transfection Chondrocyte Cell Line Chondrocytes Rheumatology Osteoarthritis Gene expression medicine Humans Pharmacology (medical) Aggrecans RNA Messenger Viability assay RNA Small Interfering Aggrecan Aged Kashin-Beck Disease Analysis of Variance Kashin–Beck disease Cartilage SOX9 Transcription Factor Middle Aged medicine.disease Molecular biology ADAM Proteins Phenotype medicine.anatomical_structure ADAMTS4 Female Matrix Metalloproteinase 3 Sulfotransferases |
| Zdroj: | Rheumatology. 50:2176-2186 |
| ISSN: | 1462-0332 1462-0324 |
| DOI: | 10.1093/rheumatology/ker230 |
| Popis: | Objective. Primary OA and KashinBeck 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-Osulphotransferase 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. |
| Databáze: | OpenAIRE |
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