Differential expression of GADD45beta in normal and osteoarthritic cartilage: potential role in homeostasis of articular chondrocytes.
Differential expression of GADD45beta in normal and osteoarthritic cartilage: potential role in homeostasis of articular chondrocytes.
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DOI：
10.1002/art.23504
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发表时间：
2008-07
影响因子：
--
通讯作者：
Goldring, Mary B.
中科院分区：
文献类型：
--
作者：
Ijiri, Kosei;Zerbini, Luiz F.;Peng, Haibing;Otu, Hasan H.;Tsuchimochi, Kaneyuki;Otero, Miguel;Dragomir, Cecilia;Walsh, Nicole;Bierbaum, Benjamin E.;Mattingly, David;van Flandern, Geoff;Komiya, Setsuro;Aigner, Thomas;Libermann, Towia A.;Goldring, Mary B.
我们的前期研究表明，生长停滞和DNA损伤诱导蛋白45β（GADD 45 β）延长了发育中的小鼠胚胎肥大软骨细胞的存活。因此，本研究旨在研究GADD 45 β是否在成人关节软骨中发挥作用。在2个独立的微阵列分析中，将晚期骨关节炎（OA）患者软骨的基因表达谱与早期OA患者和正常对照组的基因表达谱进行比较。采用Mankin评分法对软骨组织学特征进行分级，免疫组化法对GADD 45 β进行定位。用小干扰RNA（siRNA）-GADD 45 β或GADD 45 β-FLAG转导人软骨细胞。实时荧光定量RT-PCR检测GADD 45 β和COL 2A 1 mRNA水平，瞬时转染检测启动子活性。通过Hoechst 33342染色浓缩的染色质检测细胞死亡。GADD 45 β在正常供体和早期OA患者的软骨中的表达水平高于晚期OA患者的软骨。正常软骨中的所有软骨细胞核均进行GADD 45 β免疫染色。在早期OA软骨中，GADD 45 β主要分布于软骨细胞团、中深层细胞和骨赘中。相反，与早期OA或正常软骨相比，晚期OA中COL 2A 1、其他胶原基因和与骨骼发育相关的因子上调。在过表达和敲除实验中，GADD 45 β下调COL 2A 1 mRNA和启动子活性。NF-κB过表达增加了GADD 45 β启动子活性，siRNA-GADD 45 β降低了人关节软骨细胞本身的存活率，并增强了肿瘤坏死因子α诱导的细胞死亡。这些观察结果表明，GADD 45 β可能通过调节胶原基因表达和促进正常成人软骨和早期OA中的细胞存活，在调节软骨细胞稳态中发挥重要作用。
Our previous study suggested that growth arrest and DNA damage–inducible protein 45β (GADD45β) prolonged the survival of hypertrophic chondrocytes in the developing mouse embryo. This study was undertaken, therefore, to investigate whether GADD45β plays a role in adult articular cartilage. Gene expression profiles of cartilage from patients with late-stage osteoarthritis (OA) were compared with those from patients with early OA and normal controls in 2 separate microarray analyses. Histologic features of cartilage were graded using the Mankin scale, and GADD45β was localized by immunohistochemistry. Human chondrocytes were transduced with small interfering RNA (siRNA)–GADD45β or GADD45β-FLAG. GADD45β and COL2A1 messenger RNA (mRNA) levels were analyzed by real-time reverse transcriptase–polymerase chain reaction, and promoter activities were analyzed by transient transfection. Cell death was detected by Hoechst 33342 staining of condensed chromatin. GADD45β was expressed at higher levels in cartilage from normal donors and patients with early OA than in cartilage from patients with late-stage OA. All chondrocyte nuclei in normal cartilage immunostained for GADD45β. In early OA cartilage, GADD45β was distributed variably in chondrocyte clusters, in middle and deep zone cells, and in osteophytes. In contrast, COL2A1, other collagen genes, and factors associated with skeletal development were up-regulated in late OA, compared with early OA or normal cartilage. In overexpression and knockdown experiments, GADD45β down-regulated COL2A1 mRNA and promoter activity. NF-κB overexpression increased GADD45β promoter activity, and siRNA-GADD45β decreased cell survival per se and enhanced tumor necrosis factor α–induced cell death in human articular chondrocytes. These observations suggest that GADD45β might play an important role in regulating chondrocyte homeostasis by modulating collagen gene expression and promoting cell survival in normal adult cartilage and in early OA.
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影响因子：
4.8
作者：
Ijiri, K;Zerbini, LF;Goldring, MB
通讯作者：
Goldring, MB
DOI：
10.1073/pnas.011404098
发表时间：
2001-01-02
影响因子：
11.1
作者：
Li, C;Wong, WH
通讯作者：
Wong, WH
影响因子：
30.8
作者：
Kizawa, H;Kou, I;Ikegawa, S
通讯作者：
Ikegawa, S
影响因子：
2.4
作者：
Alexopoulos, LG;Williams, GM;Guilak, F
通讯作者：
Guilak, F
影响因子：
--
作者：
Aigner, T;Vornehm, SI;Bayliss, MT
通讯作者：
Bayliss, MT