USP15 通过去泛素化 TGF-βR1 体外增强增生性瘢痕衍生成纤维细胞的增殖、迁移和胶原沉积。
USP15 Enhances the Proliferation, Migration, and Collagen Deposition of Hypertrophic Scar-Derived Fibroblasts by Deubiquitinating TGF-βR1 In Vitro.
机构信息
From the Institute of Burn and Departments of Orthopedic Surgery and Nursing, The First Affiliated Hospital of Nanchang University.
出版信息
Plast Reconstr Surg. 2021 Nov 1;148(5):1040-1051. doi: 10.1097/PRS.0000000000008488.
BACKGROUND
Hypertrophic scar is a fibroproliferative disorder caused by skin injury. The incidence of hypertrophic scar following trauma or burns is 40 to 70 percent or 70 percent, respectively. It has been shown that transforming growth factor (TGF) β1/Smad signaling plays a crucial role in hypertrophic scar, and that USP15 can regulate the activity of TGFβ1/Smad signaling to affect the progression of the disease. However, the underlying mechanism of USP15 in hypertrophic scar remains unclear. The authors hypothesized that USP15 was up-regulated and enhanced the proliferation, migration, invasion, and collagen deposition of hypertrophic scar-derived fibroblasts by deubiquitinating TGF-β receptor I (TβRI) in vitro.
METHODS
Fibroblasts were isolated from human hypertrophic scars in vitro. The knockdown and overexpression of USP15 in hypertrophic scar-derived fibroblasts were performed using lentivirus infection. The effect of USP15 on hypertrophic scar-derived fibroblast proliferation, migration, and invasion, and the expression of TβRI, Smad2, Smad3, α-SMA, COL1, and COL3, were detected by Cell Counting Kit-8, scratch, invasion, quantitative real-time polymerase chain reaction, and Western blot assays. The interaction between USP15 and TβRI was detected by co-immunoprecipitation and ubiquitination assays.
RESULTS
The authors demonstrated that USP15 knockdown significantly inhibited the proliferation, migration, and invasion of hypertrophic scar-derived fibroblasts in vitro and down-regulated the expression of TβRI, Smad2, Smad3, α-SMA, COL1, and COL3; in addition, USP15 overexpression showed the opposite trends (p < 0.05). Co-immunoprecipitation and ubiquitination assays revealed that USP15 interacted with TβRI and deubiquitinated TβRI.
CONCLUSION
USP15 enhances the proliferation, migration, invasion, and collagen deposition of hypertrophic scar-derived fibroblasts by deubiquitinating TβRI in vitro.
背景
增生性瘢痕是一种由皮肤损伤引起的纤维增生性疾病。创伤或烧伤后增生性瘢痕的发生率分别为 40%至 70%或 70%。研究表明，转化生长因子（TGF）β1/Smad 信号转导在增生性瘢痕中起着至关重要的作用，USP15 可以调节 TGFβ1/Smad 信号转导的活性，从而影响疾病的进展。然而，USP15 在增生性瘢痕中的潜在机制尚不清楚。作者假设，USP15 通过体外去泛素化 TGF-β 受体 I（TβRI）而上调并增强增生性瘢痕衍生成纤维细胞的增殖、迁移、侵袭和胶原沉积。
方法
体外分离人增生性瘢痕成纤维细胞。采用慢病毒感染对增生性瘢痕衍生成纤维细胞中的 USP15 进行敲低和过表达。通过细胞计数试剂盒-8、划痕、侵袭实验、实时定量聚合酶链反应和 Western blot 检测 USP15 对增生性瘢痕衍生成纤维细胞增殖、迁移和侵袭的影响以及 TβRI、Smad2、Smad3、α-SMA、COL1 和 COL3 的表达。通过免疫共沉淀和泛素化实验检测 USP15 与 TβRI 的相互作用。
结果
作者证实，USP15 敲低显著抑制增生性瘢痕衍生成纤维细胞的体外增殖、迁移和侵袭，并下调 TβRI、Smad2、Smad3、α-SMA、COL1 和 COL3 的表达；相反，USP15 过表达则呈现出相反的趋势（p<0.05）。免疫共沉淀和泛素化实验表明，USP15 与 TβRI 相互作用并去泛素化 TβRI。
结论
USP15 通过体外去泛素化 TβRI 增强增生性瘢痕衍生成纤维细胞的增殖、迁移、侵袭和胶原沉积。
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