Cited by 108Osteoarthritis Treatment and MechanismsTelomeres, Telomerase, and SenescenceProteoglycans and glycosaminoglycans research

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Recovery of function in osteoarthritic chondrocytes induced by p16INK4a-specific siRNA in vitro

Zhou, H. W.·2004·Rheumatology

DOI 10.1093/rheumatology/keh127

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Abstract

OBJECTIVE: To demonstrate the roles of p16(INK4a) in the senescence of human chondrocytes and the progression of osteoarthritis (OA). METHODS: Immunohistochemistry and reverse transcriptase polymerase chain reaction (RT-PCR) were performed to examine p16(INK4a) expression in fetal, normal age-matched and OA cartilage, and Western blot was used in primary cultured chondrocytes from different origins. To explore a functional p16(INK4a) knockdown in OA chondrocytes, the primary cultured cells were treated with p16(INK4a)-specific small interfering ribonucleic acids (siRNAs). Expression of p16(INK4a), p14(ARF) and p53 was observed by Western blot and RT-PCR. The phosphorylation status of pRb, senescence-associated beta-galactosidase (SA-beta-gal), cell G1/S transition and cell proliferation were studied by Western blot, histological staining, 3H-thymidine incorporation and cell counts respectively. Expression of the collagen I, collagen II and aggrecan genes was measured by semiquantitative RT-PCR. To establish the response of chondrocytes to cytokines, cells were treated with transforming growth factor-beta1 (TGF-beta1) or interleukin-1alpha (IL-1alpha) and examined for incorporation of 3H-thymidine, 3H-proline and 35S-sulphate respectively. RESULTS: A significant increase of p16(INK4a) was detected in OA chondrocytes compared with normal age-matched and fetal chondrocytes (P<0.01) in vivo and in vitro. Treated with p16(INK4a)-specific siRNAs, OA chondrocytes displayed a significant decrease in p16(INK4a) expression with an increase of phosphorylated pRb, but no alteration of p14(ARF) and p53 expression, followed by decreases of senescent features and increases in the expression of some chondrocyte-specific genes and overall repair capacity. CONCLUSIONS: p16(INK4a) is instrumental in the senescence of human articular chondrocytes or OA. The reduction of p16(INK4a) by RNA interference (RNAi) contributed to the recovery of osteoarthritic chondrocytes, suggesting that p16(INK4a) may be a viable future therapeutic candidate.

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Authors: Zhou, H. W.
Affiliations: Peking University (CN)
Corresponding author: Zhou, H. W.
ORCID: Zhou, H. W.
Venue: Rheumatology
Publisher: Oxford University Press (OUP)
Published: 2004-01-27
Volume: 43
Issue: 5
Pages: 555-568
ISSN: 0263-7103 (ISSN-L)
1460-2172
DOI: 10.1093/rheumatology/keh127
OpenAlex: W2134414575
Cited by: 108
References: 65
Citations by year: 2026: 3 · 2025: 5 · 2024: 6 · 2023: 3 · 2022: 6 · 2021: 11 · 2020: 7 · 2019: 10 · 2018: 6 · 2017: 7 · 2016: 4 · 2015: 5 · 2014: 1 · 2013: 5 · 2012: 4
FWCI: 2.33
Type: journal article
Language: en
Open access: closed
Locations: British journal of rheumatology (publishedVersion)
PubMed (publishedVersion)
Topics: Health Sciences › Medicine › Rheumatology › Osteoarthritis Treatment and Mechanisms
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Telomeres, Telomerase, and Senescence
Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Proteoglycans and glycosaminoglycans research
Keywords: chondrocytes, p16INK4a, cellular senescence, articular cartilage, siRNA knockdown, RNA interference, retinoblastoma protein phosphorylation, type II collagen, aggrecan, TGF-β1, interleukin-1 alpha, chondrocyte proliferation, cartilage repair, p14ARF, p53, G1/S transition
MeSH terms: Aged, Aged, 80 and over, Cartilage, Articular, Cells, Cultured, Gene Expression Regulation, Humans, Middle Aged, Transfection, Tumor Suppressor Protein p53, Cytokines, Cellular Senescence, Disease Progression, Chondrocytes, Cyclin-Dependent Kinase Inhibitor p16, Reverse Transcriptase Polymerase Chain Reaction, Osteoarthritis, Knee, Tumor Suppressor Protein p14ARF, RNA, Small Interfering

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Recovery of function in osteoarthritic chondrocytes induced by p16INK4a-specific siRNA in vitro

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