Cadherin-11 cooperates with inflammatory factors to promote the migration and invasion of fibroblast-like synoviocytes in pigmented villonodular synovitis
Cadherin-11 cooperates with inflammatory factors to promote the migration and invasion of fibroblast-like synoviocytes in pigmented villonodular synovitis
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DOI：
10.7150/thno.48666
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发表时间：
2020-01-01
期刊：
影响因子：
12.4
通讯作者：
Ao, Yingfang
中科院分区：
文献类型：
--
作者：
Cao, Chenxi;Wu, Fei;Ao, Yingfang
基本原理：色素沉着绒毛结节性滑膜炎（PVNS）是一种发生在滑膜组织中的破坏性良性肿瘤样增生性疾病。成纤维细胞样滑膜细胞 (FLS) 是构成 PVNS 滑膜衬里层结构的主要细胞类型。由于PVNS复发率高、侵袭、迁移和软骨破坏能力强，给患者造成较大损害，手术切除的疗效并不理想。因此，迫切需要探索PVNS的发病机制并确定新的治疗靶点。目前，PVNS的发病机制尚不清楚，关于PVNS是炎症性疾病还是肿瘤性疾病尚存在不确定性和争议。 Cadherin-11 是一种经典分子，可介导 FLS 中血友病细胞间粘附，并在正常滑膜衬里层形成中发挥重要作用。本研究旨在探讨炎症和钙粘蛋白11在PVNS发病机制中的作用，并确定钙粘蛋白11作为PVNS治疗分子靶点的作用。方法：FLS主要从关节镜滑膜切除术中的PVNS患者中培养。使用人抗体阵列评估 PVNS 滑液中的细胞因子水平。通过 qPCR、Western blot、组织免疫组织化学和细胞免疫荧光检测 PVNS FLS 的 Cadherin-11 表达。在有或没有炎症因子刺激的情况下，Cadherin-11 通过 siRNA 下调或通过质粒上调，并且 LY294002 抑制 PI3K/Akt。使用 Transwell 和伤口愈合试验测试 PVNS FLS 的迁移和侵袭能力。通过蛋白质印迹检测核因子 kappaB (NF-kappa B) 和丝裂原激活蛋白激酶 (MAPK) 通路的激活。通过共培养测定评估 PVNS FLS 造成的软骨细胞损伤。结果：PVNS 患者滑液中的炎症因子（IL-1β 和 TNF-α）显着上调。 Cadherin-11在PVNS患者的FLS中高表达，并且与PVNS的复发、关节外迁移和软骨破坏呈正相关。敲除cadherin-11可抑制PVNS FLS的迁移和侵袭。此外，炎症因子上调cadherin-11的表达，激活NF-kappa B和MAPK信号通路并导致软骨破坏。钙粘蛋白-11的抑制可阻断IL-1β和TNF-α诱导的上述通路的激活、PVNS FLS的迁移和侵袭以及软骨细胞的损伤。此外，PVNS FLS中cadherin-11的表达升高以及PVNS FLS的迁移和侵袭通过抑制PI3K/Akt信号通路而下调。结论：Cadherin-11在PVNS的发病机制中发挥着重要作用，并与炎症因子形成正反馈环，进一步激活NF-kappa B和MAPK通路，引发炎症级联反应。 Cadherin-11介导的炎症导致PVNS具有高复发性、侵袭性和较强的软骨破坏能力，最终促进PVNS从最初的炎症性疾病向肿瘤性疾病转变。因此，抑制cadherin-11及其相关的炎症反应代表了PVNS的新治疗策略。
Rationale: Pigmented villonodular synovitis (PVNS) is a destructive benign tumor-like hyperplastic disease that occurs in synovial tissue. Fibroblast-like synoviocytes (FLS) are the predominant cell type comprising the structure of the PVNS synovial lining layer. Due to a high recurrence rate, high invasion, migration, and cartilage destruction ability, PVNS causes substantial damage to patients and the efficacy of surgical resection is not satisfactory. Therefore, exploring the pathogenesis and identifying novel therapeutic targets for PVNS are urgently required. Currently, the pathogenesis of PVNS remains unclear, and there is uncertainty and controversy regarding whether PVNS is an inflammatory or a neoplastic disease. Cadherin-11 is a classical molecule that mediates hemophilic cell-to-cell adhesion in FLS and plays an important role in the normal synovium lining layer formation. This study aimed to explore the role of inflammation and cadherin-11 in PVNS pathogenesis and determine the effects of cadherin-11 as a molecular target for PVNS treatment.Methods: FLS were primarily cultured from PVNS patients during arthroscopic synovectomy. The level of cytokines in the PVNS synovial fluid was evaluated using a human antibody array. Cadherin-11 expression of PVNS FLS was detected by qPCR, Western blots, tissue immunohistochemistry, and cell immunofluorescence. Cadherin-11 was down-regulated by siRNA or up-regulated with a plasmid, with or without inflammatory factor stimulation, and PI3K/Akt was inhibited with LY294002. The capacity of migration and invasion of PVNS FLS was tested using Transwell and wound-healing assays. Activation of the nuclear factor-kappaB (NF-kappa B) and mitogen-activated protein kinase (MAPK) pathways was detected by Western blots. Chondrocyte damage by PVNS FLS was assessed with a co-culture assay.Results: Inflammatory factors (IL-1 beta and TNF-alpha) in the synovial fluid of PVNS patients were significantly up-regulated. Cadherin-11 was highly expressed in the FLS of PVNS patients, and positively correlated with recurrence, extra-articular migration, and cartilage destruction of PVNS. Knocking down of cadherin-11 inhibited the migration and invasion of PVNS FLS. Moreover, inflammatory factors up-regulated the expression of cadherin-11, which activated the NF-kappa B and MAPK signaling pathways and led to cartilage destruction. Inhibition of cadherin-11 blocked IL-1 beta- and TNF-alpha-induced activation of the above pathways, migration and invasion of PVNS FLS, and damage of chondrocyte. In addition, the elevation of cadherin-11 expression, together with the migration and invasion, of PVNS FLS was down-regulated by the inhibition of the PI3K/Akt signaling pathway.Conclusions: Cadherin-11 plays an important role in the pathogenesis of PVNS and forms a positive feedback loop with inflammatory factors, which further activates the NF-kappa B and MAPK pathways to trigger an inflammatory cascade. Cadherin-11-mediated inflammation results in PVNS with high recurrence, invasiveness, and strong cartilage destruction ability, and eventually promotes the transformation of PVNS from the initial inflammatory disease to neoplastic disease. Thus, inhibition of cadherin-11 together with its related inflammatory reaction, represents a new therapeutic strategy for PVNS.