Varp通过Tyrp1影响黑色素瘤的发生发展及其调控机制研究

Malignant melanoma（MM）is an aggressive, therapy-resistant malignancy of melanocytes. Currently, the pathogenesis of MM remained unclear, and there was still no effective clinical treatment for it. It is critical to investigate the pathogenesis of MM and find a new treatment for it. Tyrp1 is an important regulator involved in MM progression. Intracellular transport of Tyrp1 is regulated by a novel protein Varp, whose function is largely indistinct. In our previous research, we have built an MM model B16/Varpi, in which Varp was knocked-down by specific shRNA. We demonstrated that silencing of Varp inhibits both Tyrp1 expression and cell proliferation in B16.And inhibition of Tyrp1 disappeared when treated cell with proteasome inhibitor MG132. Preliminary experiment on nude mice showed that the tumorigenicity of B16/Varpi was less than B16/Mock. All of these indicated that Varp might play an key role in regulating MM through Tyrp1. This project will verify that Varp affects cell proliferation, migration and invasion in vitro, as well as tumor growth and metastasis in nude mice. Further more, we will demonstrate that Varp regulates the pathogenesis and progression of MM through Tyrp1 with several means. This project may provide new direction for the work on research of the mechanism and the treatmant, molecular targeting therapy, of MM.

恶性黑色素瘤（malignant melanoma，MM）是一种原发于皮肤的恶性肿瘤。Tyrp1蛋白是黑色素细胞恶变过程中重要的调节因子，其在细胞中的转运受Varp蛋白的调节。我们前期研究证明Varp表达沉默可抑制MM细胞中Tyrp1的表达，降低MM在体外的增殖能力及在体内的成瘤能力。这些结果提示Varp可能通过Tyrp1对MM生长起调节作用。本项目拟新建Tyrp1表达沉默的MM细胞模型，与已有细胞模型共同利用，在体外检测Varp通过Tyrp1对MM细胞生长、迁移、侵袭的影响；建立裸鼠成瘤及转移模型，在体内检测Varp通过Tyrp1对MM成瘤以及转移的影响。本项目通过多种实验手段证明Varp通过Tyrp1影响MM的发生发展，并对其调控机制进行初步探索，为阐明MM发病机理、探索MM分子靶向治疗提供新的线索。

恶性黑色素瘤（malignant melanoma，MM）是一种原发于皮肤的恶性肿瘤。Tyrp1蛋白是黑色素细胞恶变过程中重要的调节因子，其在细胞中的转运受Varp蛋白的调节。本项目旨在利用小鼠黑色素瘤细胞B16建立细胞模型，对Varp和Tyrp1在黑色素瘤发生发展中的调节作用进行探索研究。主要研究内容是分别建立Varp和Tyrp1表达沉默的MM细胞模型，在体外运用细胞增殖实验、细胞迁移实验和细胞侵袭实验检测Varp通过Tyrp1对MM细胞生长、迁移、侵袭的影响，并初步检测细胞迁移侵袭相关蛋白表达的变化；建立裸鼠成瘤及转移模型，在体内检测Varp和Tyrp1对MM成瘤以及转移的影响；收集人类MM癌组织，检测Varp与Tyrp1蛋白在癌及癌旁组织中的表达差异。在项目执行过程中，取得了以下成果：(1)分别建立Varp表达沉默的MM细胞模型（B16/Varpi）和Tyrp1表达沉默的MM细胞模型（B16/Tyrp1i），为今后进一步研究Varp及Tyrp1在MM细胞中的功能提供基础。(2)与对照细胞相比，B16/Varpi和B16/Tyrp1i的体外增殖与迁移能力下降，细胞迁移及侵袭相关蛋白MMP-2表达降低，Transwell实验证明，与B16/Varpi相比，B16/Tyrp1i的侵袭能力显著降低。（3）与对照细胞相比，B16/Varpi和B16/Tyrp1i的体内成瘤能力和转移能力有所降低。(4)Varp与Tyrp1蛋白在人类MM癌组织中的表达较癌旁组织明显为高。本项目取得的研究成果，为阐明MM发病机理、探索MM分子靶向治疗提供了新的线索。