Experimental Oncology Unit, Department of Microbiology, Immunology and Parasitology, University of São Paulo, São Paulo, Brazil.
Recepta Biopharma São Paulo, Brazil.
Sci Rep. 2017 Jan 12;7:40585. doi: 10.1038/srep40585.
Silencing of SOCS1 protein with shRNAi lentivirus (shR-SOCS1) led to partial reversion of the tumorigenic phenotype of B16F10-Nex2 melanoma cells. SOCS1 silencing inhibited cell migration and invasion as well as in vitro growth by cell cycle arrest at S phase with increased cell size and nuclei. Down-regulation of SOCS1 decreased the expression of epidermal growth factor receptor, Ins-Rα, and fibroblast growth factor receptors. The present work aimed at analyzing the SOCS1 cell signaling and expression of proteins relevant to tumor development. An RNA microarray analysis of B16F10-Nex2 melanoma cells with SOCS1 silenced by shRNAi-SOCS1 was undertaken in comparison with cells transduced with the empty vector. Among 609 differentially expressed genes, c-Kit, Met and EphA3 cytokine/tyrosine-kinase (TK) receptors were down regulated. A significant decrease in the expression of TK receptors, the phosphorylation of mediators of ERK1/2 and p38 pathways and STAT3 (S727) were observed. Subcutaneous immunization with shR-SOCS1-transduced viable tumor cells rendered protection against melanoma in a syngeneic model, with decreased expression of PD-L1 and of matrix metallo-proteinases (MMPs) and CD-10 in those cells. The present work shows the role of SOCS1 in murine melanoma development and the potential of SOCS1-silenced tumor cells in raising an effective anti-melanoma immune response.
利用 shRNAi 慢病毒沉默 SOCS1 蛋白（shR-SOCS1）导致 B16F10-Nex2 黑色素瘤细胞的致瘤表型部分逆转。SOCS1 沉默通过 S 期细胞周期阻滞抑制细胞迁移和侵袭以及体外生长，同时增加细胞大小和细胞核。SOCS1 的下调降低了表皮生长因子受体、Ins-Rα 和成纤维细胞生长因子受体的表达。本研究旨在分析 SOCS1 细胞信号转导和与肿瘤发展相关的蛋白质表达。通过 RNA 微阵列分析，比较 shRNAi-SOCS1 沉默 SOCS1 的 B16F10-Nex2 黑色素瘤细胞与转导空载体的细胞。在 609 个差异表达基因中，c-Kit、Met 和 EphA3 细胞因子/酪氨酸激酶（TK）受体下调。观察到 TK 受体表达、ERK1/2 和 p38 通路以及 STAT3（S727）磷酸化的调节剂显著减少。用 shR-SOCS1 转导的活肿瘤细胞进行皮下免疫接种，在同基因模型中对黑色素瘤具有保护作用，这些细胞中 PD-L1 和基质金属蛋白酶（MMPs）和 CD-10 的表达降低。本研究表明 SOCS1 在小鼠黑色素瘤发展中的作用以及沉默 SOCS1 的肿瘤细胞在引发有效抗黑色素瘤免疫反应中的潜力。
