Jiang Shan, Zheng Ziang, Yuan Bo, Yan Rushan, Yao Qijun, Chen Haoran, Zhang Yongxun, Lei Yue, Liang Haidong
Department of Bone and Soft Tissue Repair and Reconstructive Surgery, The Second Hospital of Dalian Medical University, Dalian, China.
Transl Cancer Res. 2024 Jun 30;13(6):2847-2859. doi: 10.21037/tcr-23-2035. Epub 2024 Jun 27.
Osteosarcoma (OS) is a malignancy originating from mesenchymal tissue. Microfibril-associated protein 2 () plays a crucial role in cancer, notably promoting epithelial-mesenchymal transition (EMT). However, its involvement in OS remains unexplored.
was silenced in U2OS cells using shRNA targeting (sh-MFAP2) and validated by quantitative real-time polymerase chain reaction (qRT-PCR). We extracted gene chip data of from multiple databases (GSE28424, GSE42572, and GSE126209). Correlation analyses between and the Notch1 pathway identified through the gene set variation analysis (GSVA) enrichment analysis were conducted using the Pearson correlation method. Cellular behaviors (viability, migration, and invasion) were assessed via the Cell Counting Kit-8 (CCK-8), wound healing, and Transwell assays. EMT markers (N-cadherin, vimentin, and β-catenin) and Notch1 levels were examined by western blotting and qRT-PCR. Cell morphology was observed microscopically to evaluate EMT. Finally, the role of in OS was validated through a xenograft tumor model.
OS cell lines exhibited higher mRNA expression than normal osteoblasts. knockdown in U2OS cells significantly reduced viability, migration, and invasion, along with downregulation of N-cadherin and vimentin, as well as upregulation of β-catenin. significantly correlated with the Notch1 pathway in OS and its knockdown inhibited Notch1 protein expression. Furthermore, Notch1 activation reversed the inhibitory effects of knockdown on the malignant characteristic of U2OS cells. Additionally, knockdown inhibited tumor growth, expression levels of EMT markers, and Notch1 expression in OS tumor tissues.
Our study revealed that was an upstream regulator of the Notch1 signaling pathway to promote EMT in OS. These findings suggested as a potential OS therapy target.
骨肉瘤（OS）是一种起源于间充质组织的恶性肿瘤。微原纤维相关蛋白2（MFAP2）在癌症中起关键作用，尤其促进上皮-间质转化（EMT）。然而，其在骨肉瘤中的作用仍未得到探索。
使用靶向MFAP2的短发夹RNA（sh-MFAP2）在U2OS细胞中沉默MFAP2，并通过定量实时聚合酶链反应（qRT-PCR）进行验证。我们从多个数据库（GSE28424、GSE42572和GSE126209）中提取了MFAP2的基因芯片数据。使用Pearson相关方法对通过基因集变异分析（GSVA）富集分析确定的MFAP2与Notch1信号通路之间进行相关性分析。通过细胞计数试剂盒-8（CCK-8）、伤口愈合和Transwell实验评估细胞行为（活力、迁移和侵袭）。通过蛋白质免疫印迹和qRT-PCR检测EMT标志物（N-钙黏蛋白、波形蛋白和β-连环蛋白）以及Notch1水平。通过显微镜观察细胞形态以评估EMT。最后，通过异种移植肿瘤模型验证MFAP2在骨肉瘤中的作用。
骨肉瘤细胞系的MFAP2 mRNA表达高于正常成骨细胞。U2OS细胞中MFAP2敲低显著降低了活力、迁移和侵袭能力，同时N-钙黏蛋白和波形蛋白下调，β-连环蛋白上调。MFAP2在骨肉瘤中与Notch1信号通路显著相关，其敲低抑制了Notch1蛋白表达。此外，Notch1激活逆转了MFAP2敲低对U2OS细胞恶性特征的抑制作用。另外，MFAP2敲低抑制了骨肉瘤肿瘤组织中的肿瘤生长、EMT标志物表达水平和Notch1表达。
我们的研究表明，MFAP2是Notch1信号通路的上游调节因子，可促进骨肉瘤中的EMT。这些发现提示MFAP2作为骨肉瘤潜在的治疗靶点。
