Department of Plastic Surgery, The First Affiliated Hospital, College of Medicine, Zhejiang University, Zhejiang, China.
Department of Dermatology, The First Affiliated Hospital, College of Medicine, Zhejiang University, Zhejiang, China.
Cell Biochem Funct. 2020 Jul;38(5):604-612. doi: 10.1002/cbf.3515. Epub 2020 Feb 24.
Melanoma is a highly malignant and is a life-threatening disease with no effective treatment currently. This study aims to evaluate the significance of TUSC3, an endoplasmic reticulum stress (ERS)-inducible gene and explore its relationship with AKT/GSK3-β/β-catenin signalling pathway in melanoma cell WM451. We investigated TUSC3 expression in melanoma cell by qRT-PCR, CCK-8 and clonal formation assays were utilized to evaluate cell proliferation. Wound healing and transwell experiments detected cell migration and invasion. Flow cytometry detected the level of apoptosis. Western blot analysed MMP2, MMP9, p-AKT, p-GSK3-β, β-catenin and AKT, GSK3-β, ERS-related proteins and apoptosis-related proteins in WM451 cells. The results revealed that TUSC3 was remarkably decreased in melanoma cell lines. Overexpression of TUSC3 significantly inhibits melanoma cell WM451 biological functions and promotes expression of ERS-related proteins in WM451 cells, increases ERS in WM451 cells by inhibiting AKT/GSK3-β/β-catenin pathway. These finding suggest that TUSC3 regulates biological functions of melanoma cells WM451 and increases ERS in melanoma cells WM451 via the inhibition of the AKT/GSK3-β/β-catenin signalling pathway. SIGNIFICANCE OF THE STUDY: Melanoma is a highly malignant and is a life-threatening disease with no effective treatment currently. Therefore, studying the molecular mechanism of melanoma occurrence and metastasis is essential for the treatment of melanoma. Meanwhile, mounting studies suggest that TUSC3 is considered to be closely associated with the development of various malignancies. TUSC3 regulates proliferation, migration and epithelial-to-mesenchymal transition, but the molecular mechanism of the tumour suppressor effects of TUSC3 on melanoma cells is not well understood. Our study demonstrates that TUSC3 inhibits biological function of melanoma cells and increases ERS in melanoma cells by inhibiting AKT/GSK3-β/β-catenin pathway. And this is expected to be a new target and method for the treatment of melanoma.
黑色素瘤是一种高度恶性的疾病，目前尚无有效的治疗方法，危及生命。本研究旨在评估内质网应激（ERS）诱导基因 TUSC3 的意义，并探讨其与黑色素瘤细胞 WM451 中 AKT/GSK3-β/β-catenin 信号通路的关系。我们通过 qRT-PCR 研究了黑色素瘤细胞中 TUSC3 的表达，通过 CCK-8 和克隆形成实验评估细胞增殖，通过划痕愈合和 Transwell 实验检测细胞迁移和侵袭，通过流式细胞术检测细胞凋亡水平，通过 Western blot 分析 WM451 细胞中 MMP2、MMP9、p-AKT、p-GSK3-β、β-catenin 和 AKT、GSK3-β、ERS 相关蛋白和凋亡相关蛋白。结果表明，黑色素瘤细胞系中 TUSC3 显著降低。过表达 TUSC3 显著抑制黑色素瘤细胞 WM451 的生物学功能，并促进 WM451 细胞中 ERS 相关蛋白的表达，通过抑制 AKT/GSK3-β/β-catenin 通路增加 WM451 细胞中的 ERS。这些发现表明 TUSC3 通过抑制 AKT/GSK3-β/β-catenin 信号通路调节 WM451 黑色素瘤细胞的生物学功能，并增加 WM451 黑色素瘤细胞中的 ERS。研究意义：黑色素瘤是一种高度恶性的疾病，目前尚无有效的治疗方法，危及生命。因此，研究黑色素瘤发生和转移的分子机制对于治疗黑色素瘤至关重要。同时，越来越多的研究表明，TUSC3 被认为与各种恶性肿瘤的发生密切相关。TUSC3 调节增殖、迁移和上皮间质转化，但 TUSC3 对黑色素瘤细胞的肿瘤抑制作用的分子机制尚不清楚。我们的研究表明，TUSC3 通过抑制 AKT/GSK3-β/β-catenin 通路抑制黑色素瘤细胞的生物学功能并增加黑色素瘤细胞中的 ERS。这有望成为治疗黑色素瘤的新靶点和新方法。
