|Recovery of function in osteoarthritic chondrocytes induced by p16(INK4a)-specific siRNA in vitro|
|Zhou, HW; Lou, SQ; Zhang, K|
|关键词||p16(INK4a) senescence osteoarthritis siRNA|
|WOS标题词||Science & Technology|
|关键词[WOS]||RAT ARTICULAR CHONDROCYTES ; INDUCED PREMATURE SENESCENCE ; ANTERIOR CRUCIATE LIGAMENT ; NORMAL HUMAN FIBROBLASTS ; EPIDERMAL GROWTH-FACTOR ; CELL-CYCLE ; LIFE-SPAN ; REPLICATIVE SENESCENCE ; TELOMERASE ACTIVITY ; RNA INTERFERENCE|
Objective. To demonstrate the roles of p(16INK4a) 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 p(16INK4a) 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 p(16INK4a) 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 G(1)/S transition and cell proliferation were studied by Western blot, histological staining, H-3-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-lalpha (IL-1alpha) and examined for incorporation of H-3-thymidine, H-3-proline and S-35-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.
|作者单位||Peking Univ, Dept Orthopaed Surg, Hosp 3, Beijing 100083, Peoples R China|
|Zhou, HW,Lou, SQ,Zhang, K. Recovery of function in osteoarthritic chondrocytes induced by p16(INK4a)-specific siRNA in vitro[J]. RHEUMATOLOGY,2004,43(5):555-568.|
|APA||Zhou, HW,Lou, SQ,&Zhang, K.(2004).Recovery of function in osteoarthritic chondrocytes induced by p16(INK4a)-specific siRNA in vitro.RHEUMATOLOGY,43(5),555-568.|
|MLA||Zhou, HW,et al."Recovery of function in osteoarthritic chondrocytes induced by p16(INK4a)-specific siRNA in vitro".RHEUMATOLOGY 43.5(2004):555-568.|