The specific RIP1 inhibitor necrostatin-1 ameliorated degradation of ECM in human SW1353 cells

Abstract

Abnormal destruction of the components of the articular extracellular matrix (ECM) such as type II collagen and aggrecan caused by advanced glycation end products (AGEs) has been considered as one of the pathological characteristics of osteoarthritis (OA). Receptor-interacting protein 1 (RIP1), an important serine/threonine kinase, possesses a variety of biological functions including cell proliferation, survival and death. The physiological roles of RIP1 in OA have not been reported before. Here, we found that AGEs increased the expression of RIP1 in human chondrosarcoma cell line SW1353 cells. Importantly, we found that antagonism of RIP1 using its specific inhibitor necrostatin-1 (Nec-1) ameliorated AGE-induced degradation of type II collagen and aggrecan in SW1353 cells. We also found that treatment with Nec-1 reduced the expression of MMP-3 and MMP-13 but restored the expression of Tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2. Also, our results indicate that Nec-1 inhibited AGE-induced expression of ADAMTS-4 and ADAMTS-5. Mechanistically, we found that Nec-1 treatment inhibited the activation of JNK and the transcriptional factor AP-1 by reducing the expressions of c-Fos and c-Jun, the two main components of AP-1. Additionally, we found that Nec-1 treatment abolished AGE-induced activation of the transcriptional factor NF-jB by suppressing the nuclear translocation of p65. These findings suggest that RIP1 might be an important therapeutic target of OA.

A long non-coding RNA, HOTAIR, promotes cartilage degradation in osteoarthritis by inhibiting WIF-1 expression and activating Wnt pathway

“…To understand the link between HOTAIR and OArelated gene expression, we studied the effects of HOTAIR silencing and over-expression in chondrocytes by using SW1353 cells as an established chondrocytic cell model and a reliable transfection host [42]. Although SW1353 is a chondrosarcoma cell line, the cells exhibit similar responses as primary chondrocytes to a variety of catabolic cytokines relevant to OA, and have been used as the cell line of choice in many studies to investigate the molecular mechanisms of OA pathogenesis [43][44][45]. We found that HOTAIR over-expression in SW1353 cells matched the expression profile of OA-related genes in human primary osteoarthritic chondrocytes, characterised by the upregulation of catabolic enzymes (MMP-13, ADAMTS5) and cartilage repair markers (BMP-2), and downregulation of anabolic markers (TIMP3, SOX9) and cartilage matrix proteins (ACAN).…”

MicroRNA-1 Modulates Chondrocyte Phenotype by Regulating FZD7 of Wnt/ β-Catenin Signaling Pathway

“…To elucidate the link between miR-1 expression and OA-associated gene transcription, we studied the effects of miR-1 over-expression and under-expression in chondrocytes by transfecting SW1353 cells with a miR-1 agonist and antagonist, respectively. Although SW1353 is a chondrosarcoma cell line with limited similarities in gene expression profile compared to primary human chondrocytes, they have similar responses as primary chondrocytes to catabolic cytokines such as IL-1β, and have been used in many studies to investigate protease expression and regulation in chondrocytic cells [34], or molecular mechanisms of pathogenesis in OA [35][36][37]. In our study, we chose SW1353 cells for use as a reliable transfection host that replicated some of the key characteristics of chondrocytic cells.…”

A long non-coding RNA, HOTAIR, promotes cartilage degradation in osteoarthritis by inhibiting WIF-1 expression and activating Wnt pathway

“…To understand the link between HOTAIR and OA-related gene expression, we studied the effects of HOTAIR silencing and over-expression in chondrocytes by using SW1353 cells as an established chondrocytic cell model and a reliable transfection host (42). Although SW1353 is a chondrosarcoma cell line, the cells exhibit similar responses as primary chondrocytes to a variety of catabolic cytokines relevant to OA, and have been used as the cell line of choice in many studies to investigate the molecular mechanisms of OA pathogenesis (43)(44)(45). We found that HOTAIR over-expression in SW1353 cells matched the expression pro le of OA-related genes in human primary osteoarthritic chondrocytes, The roles of WIF-1 in skeletal tissue development has been investigated in other studies.…”