Institute of Biochemistry and Molecular Biology of Guangdong Medical University, No. 2 Wenming Dong Road, Xiashan District, Zhanjiang, 524023, Guangdong, China.
Department of Clinical Laboratory of Zhanjiang Central Hospital, Zhanjiang, 524023, China.
Cell Mol Biol Lett. 2020 Nov 3;25(1):49. doi: 10.1186/s11658-020-00240-x.
AIM: To explore the effect of miR-296-5p on the metastasis of nasopharyngeal carcinoma (NPC) cells and investigate the underlying mechanism. METHODS: The expressions of miR-296-5p in NPC tissues and cells were determined using GSE32920 database analysis and real-time PCR and miRNA microarray assays. An miR-296-5p mimic and inhibitor were transfected into NPC cells. Then, immunofluorescence imaging, scratch wound-healing, transwell migration and invasion assays were used to observe the effects of miR-296-5p on cell metastasis and invasion. Real-time PCR and western blotting were carried out to detect the expressions of genes and proteins related to epithelial-mesenchymal transition (EMT). A dual luciferase reporter assay was used to identify whether TGF-β is the target gene of miR-296-5p. Finally, TGF-β expression plasmids were transfected into NPC cells to verify the role of TGF-β in the miR-296-5p-mediated inhibition of nasopharyngeal carcinoma cell metastasis. RESULTS: Our results show that miR-296-5p inhibits the migratory and invasive capacities of NPC cells by targeting TGF-β, which suppresses EMT. Importantly, the miR-296-5p level was significantly lower in human NPC tissues than in adjacent normal tissues. It also negatively correlated with TGF-β and was significantly associated with the lymph node metastasis of patients with NPC. CONCLUSIONS: Our findings show that miR-296-5p represses the EMT-related metastasis of NPC by targeting TGF-β. This provides new insight into the role of miR-296-5p in regulating NPC metastasis and invasiveness.
目的：探讨 miR-296-5p 对鼻咽癌（NPC）细胞转移的影响，并探讨其潜在机制。
方法：通过 GSE32920 数据库分析和实时 PCR 及 miRNA 微阵列分析检测 NPC 组织和细胞中 miR-296-5p 的表达。转染 miR-296-5p 模拟物和抑制剂到 NPC 细胞中。然后，通过免疫荧光成像、划痕愈合、Transwell 迁移和侵袭实验观察 miR-296-5p 对细胞转移和侵袭的影响。实时 PCR 和 Western blot 检测上皮-间充质转化（EMT）相关基因和蛋白的表达。双荧光素酶报告实验鉴定 TGF-β 是否为 miR-296-5p 的靶基因。最后，转染 TGF-β 表达质粒到 NPC 细胞中，验证 TGF-β 在 miR-296-5p 介导的抑制鼻咽癌细胞转移中的作用。
结果：我们的结果表明，miR-296-5p 通过靶向 TGF-β 抑制 NPC 细胞的迁移和侵袭能力，从而抑制 EMT。重要的是，miR-296-5p 在人 NPC 组织中的水平明显低于相邻正常组织。它还与 TGF-β 呈负相关，与 NPC 患者的淋巴结转移显著相关。
结论：我们的研究结果表明，miR-296-5p 通过靶向 TGF-β 抑制 NPC 的 EMT 相关转移。这为 miR-296-5p 在调节 NPC 转移和侵袭中的作用提供了新的见解。
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