USP2 promotes cell proliferation and metastasis in choroidal melanoma via stabilizing Snail,Journal of Cancer Research and Clinical Oncology

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USP2 promotes cell proliferation and metastasis in choroidal melanoma via stabilizing Snail
Journal of Cancer Research and Clinical Oncology ( IF 2.7 ) Pub Date : 2023-05-18 , DOI: 10.1007/s00432-023-04855-3
Chao Wei ₁ , Xiaofei Zhao ₂ , Han Zhang ₂ , Lijuan Wang ₃

Affiliation

Background

Choroidal melanoma (CM) is an intraocular tumor that arises from melanocytes. While ubiquitin-specific protease 2 (USP2) modulates the progression of numerous diseases, its role in CM is not known. This study aimed to determine the role of USP2 in CM and elucidate its molecular mechanisms.

Methods

MTT, Transwell, and wound-scratch assays were used to investigate the function of USP2 in the proliferation and metastasis of CM. Western blotting and qRT-PCR were used to analyze the expression of USP2, Snail, and factors associated with the epithelial-mesenchymal transition (EMT). The relationship between USP2 and Snail was explored by co-immunoprecipitation and in vitro ubiquitination assays. A nude mouse model of CM was established for verifying the role of USP2 in vivo.

Results

USP2 overexpression promoted proliferation and metastasis, and induced the EMT in CM cells in vitro, while specific inhibition of USP2 by ML364 produced the opposite effects. ML364 also suppressed CM tumor growth in vivo. Mechanistically, USP2 is known to deubiquitinate Snail, stabilizing the latter through the removal of its K48 poly-ubiquitin chains. However, a catalytically inactive form of USP2 (C276A) had no effect on Snail ubiquitination and failed to increase Snail protein expression. The C276A mutant was also unable to promote CM cell proliferation, migration, and invasion, as well as EMT progression. Furthermore, Snail overexpression partly counteracted the effects of ML364 on proliferation and migration, while rescuing the effects of the inhibitor on the EMT.

Conclusions

The findings demonstrated that USP2 modulated CM development through the stabilization of Snail and suggest that USP2 may be a useful target for the development of novel treatments for CM.

中文翻译：

USP2 通过稳定 Snail 促进脉络膜黑色素瘤的细胞增殖和转移

背景

脉络膜黑色素瘤（CM）是一种起源于黑色素细胞的眼内肿瘤。虽然泛素特异性蛋白酶 2 （USP2）可调节多种疾病的进展，但其在 CM 中的作用尚不清楚。本研究旨在确定 USP2 在 CM 中的作用并阐明其分子机制。

方法

采用 MTT 、 Transwell 和伤口划痕法研究 USP2 在 CM 增殖和转移中的作用。Western blotting 和 qRT-PCR 分析 USP2 、 Snail 和上皮-间质转化（EMT）相关因子的表达。通过免疫共沉淀和体外泛素化测定探讨 USP2 与 Snail 之间的关系。建立了 CM 的裸鼠模型以验证 USP2 在体内的作用。

结果

USP2 过表达促进增殖和转移，并在体外诱导 CM 细胞中的 EMT，而 ML364 对 USP2 的特异性抑制产生相反的效果。ML364 还在体内抑制 CM 肿瘤生长。从机制上讲，已知 USP2 可以使 Snail 去泛素化，通过去除其 K48 多泛素链来稳定后者。然而，一种催化失活形式的 USP2 （C276A）对 Snail 泛素化没有影响，并且未能增加 Snail 蛋白表达。C276A 突变体也无法促进 CM 细胞增殖、迁移和侵袭，以及 EMT 进展。此外，Snail 过表达部分抵消了 ML364 对增殖和迁移的影响，同时挽救了抑制剂对 EMT 的影响。