Liu Zi-Mei, Yuan Ying, Jin Lei
Department of Oncology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200336, China.
Department of Rheumatology and Immunology, Tongren Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200336, China.
Discov Oncol. 2024 Mar 21;15(1):82. doi: 10.1007/s12672-024-00943-z.
Synovial Sarcoma (SS), a highly malignant mesenchymal neoplasm, typically carries a grim prognosis for patients presenting with high-grade or metastatic disease. Although Anlotinib, a new agent for treating soft tissue sarcomas, holds promise, its underlying mechanism remains incompletely understood. This investigation aims to delineate Anlotinib's anticancer effectiveness and potential mechanistic underpinnings in patients suffering from advanced, refractory SS.
Employing microarray assay, we examined the potential downstream targets of Anlotinib in SS therapy. A shRNA-based high-content screening was performed to identify candidate genes with the greatest influence on SW982 cell proliferation. The knockdown efficacy of selected genes within SW982 cells was confirmed using RT-qPCR as well as western blot analysis. To assess the effect of putative downstream elimination of genes with synovial sarcoma cells, cell proliferation, and apoptotic assays were carried out. Gene chip microarray as well as bioinformatics techniques were utilized to scrutinize potential signaling networks associated with the candidate downstream gene.
QPCR verified high expression of FAM83D in SW982 cells, shRNA was designed to silence FAM83D by lentivirus transfection, apoptosis assay, and cell cycle arrest showing that FAM83D downregulation augments apoptosis in SW982 cells and arrests cell cycle progression in the S stage. Inhibition of FAM83D expression upregulated STAT1 while downregulated BIRC5, MCM2, and CDK1 genes in vitro.
This experimental study identified FAM83D as a critical regulator that contributes to the proliferation and progression of SS, suggesting that FAM83D-regulated signaling pathway may serve as a prospective target in SS management.
滑膜肉瘤（SS）是一种高度恶性的间充质肿瘤，对于患有高级别或转移性疾病的患者而言，其预后通常较差。尽管安罗替尼作为一种治疗软组织肉瘤的新药具有前景，但其潜在机制仍未完全明确。本研究旨在阐明安罗替尼在晚期难治性SS患者中的抗癌效果及潜在机制基础。
采用微阵列分析，我们检测了安罗替尼在SS治疗中的潜在下游靶点。进行基于短发夹RNA（shRNA）的高内涵筛选，以鉴定对SW982细胞增殖影响最大的候选基因。使用逆转录定量聚合酶链反应（RT-qPCR）以及蛋白质免疫印迹分析来确认SW982细胞内所选基因的敲低效果。为评估滑膜肉瘤细胞中假定的下游基因消除的影响，进行了细胞增殖和凋亡检测。利用基因芯片微阵列以及生物信息学技术来研究与候选下游基因相关的潜在信号网络。
定量聚合酶链反应（QPCR）证实FAM83D在SW982细胞中高表达，设计shRNA通过慢病毒转染使FAM83D沉默，凋亡检测和细胞周期阻滞表明FAM83D下调可增强SW982细胞的凋亡并使细胞周期在S期停滞。体外抑制FAM83D表达可上调信号转导和转录激活因子1（STAT1），同时下调凋亡抑制蛋白5（BIRC5）、微小染色体维持蛋白2（MCM2）和细胞周期蛋白依赖性激酶1（CDK1）基因。
本实验研究确定FAM83D是促进SS增殖和进展的关键调节因子，提示FAM83D调节的信号通路可能成为SS治疗的潜在靶点。
