目的：探究三结构域蛋白59（TRIM59）调控人皮肤黑色素瘤细胞SK-MEL-2增殖、细胞周期、凋亡及迁移侵袭的作用机制，及其与Bcl2相关转录因子1（BCLAF1)之间的关系。方法：qPCR和WB法检测人表皮黑色素细胞HEMn-LP、人皮肤黑色素瘤细胞SK-MEL-2、UACC903、A375及36例邢台市人民医院2019年2月至2021年7月收集的皮肤黑色素瘤组织中TRIM59的mRNA和蛋白表达，使用脂质体将si-con、si-TRIM59转染至SK-MEL-2细胞中，WB法检测干扰TRIM59表达对细胞中周期蛋白D1（CCND1）、细胞周期素依赖性激酶2（CDK2）、肿瘤抑制蛋白基因（TP53）和 BCLAF1 蛋白表达的影响，CCK-8法、流式细胞术、划痕愈合实验、Transwell实验检测对细胞的活性、凋亡、迁移和侵袭的影响，免疫共沉淀（Co-IP）实验检测对细胞中TRIM59蛋白与BCLAF1结合能力的影响。结果：与HEMn-LP细胞相比，SK-MEL-2、UACC903、A375细胞中TRIM59 mRNA和TRIM59、BCLAF1蛋白均呈高表达（均P<0.05），SK-MEL-2细胞中TRIM59表达水平最高。相较于si-con组和Normal组，沉默TRIM59后，SK-MEL-2细胞的活性显著降低，细胞周期阻滞于G2期，CCND1、CDK2的蛋白表达显著降低，TP53蛋白和细胞凋亡率均显著升高，划痕抑制率明显升高，迁移侵袭细胞数明显降低（均P<0.05）。免疫共沉淀实验结果显示，TRIM59与BCLAF1之间存在蛋白结合关系。TRIM59与 BCLAF1 在肿瘤组织中的表达呈显著的正相关（r=0.878，P<0.001）。结论：干扰TRIM59表达能够抑制人皮肤黑色素瘤SK-MEL-2细胞的增殖、迁移和侵袭而促进凋亡，抑制SK-MEL-2细胞的恶性生物学行为，其机制可能与TRIM59结合BCLAF1有关。

Objective: To explore the mechanism of tripartite motif-containing 59 (TRIM59) regulating the proliferation, cell cycle,apoptosis, migration and invasion of human skin melanoma cells SK-MEL-2, and its relationship with Bcl2-associated transcription factor (BCLAF1). Methods: qPCR and WB assay were used to measure the mRNA and protein expression of TRIM59 in human epidermal melanocytes HEMN-LP, human skin melanoma cells SK-MEL-2, UACC903, A375, and 36 cases of human skin melanoma tissues collected from February 2019 to July 2021 in Xingtai people's Hospital. Si-con and si-TRIM59 were transfected into SK-MEL-2 cells using liposomes. WB assay was used to detect the effects of interference with the expression of TRIM59 on cyclin D1 (CCND1),cyclin-dependent kinase 2 (CDK2), tumor suppressor protein gene (TP53) and BCLAF1 protein expression. CCK-8 assay, flow cytometry, scratch test and Transwell test were used to detect cell activity, apoptosis, migration and invasion. The binding ability of TRIM59 protein and BCLAF1 was detected by Co-IP assay. Results: Compared with the HEMN-LP group, the mRNA and protein expression of TRIM59 BCLAF1 protein in SK-MEL-2, UACC903and A375 cells were significantly increased (P<0.05). The expression level of TRIM59 in SK-MEL-2 cells were the highest. Compared with the si-con group and the Normal group, after silencing TRIM59,the activity of SK-MEL-2 cells was significantly decreased; the G2 phase of the cell cycle was blocked; the protein expressions of CCND1 and CDK2 were decreased significantly; the TP53 protein and apoptosis rate were significantly increased; the scratch inhibition rate was significantly increased, and the numbers of migration and invasion cells were significantly decreased. (all P<0.05).The results of co-immunoprecipitation experiments showed that there was a protein-binding relationship between TRIM59 and BCLAF1. There was a significant positive correlation between TRIM59 and BCLAF1 expression in tumor tissues (r=0.878, P<0.001).Conclusion: Silencing TRIM59 expression could inhibit the proliferation, migration and invasion of skin cutaneous melanoma cells,promote apoptosis and inhibit the malignant biological behaviors of SK-MEL-2 cells. The mechanism may be related to the binding of TRIM59 with BCLAF1.

邢台市重点研发计划资助项目（No. 2020ZC233）