敲低 MTHFD2 通过 ERK 信号通路抑制鼻咽癌细胞的增殖和迁移。
Knockdown of MTHFD2 inhibits proliferation and migration of nasopharyngeal carcinoma cells through the ERK signaling pathway.
机构信息
Department of Otorhinolaryngology, Head and Neck Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China.
Department of Otorhinolaryngology, Head and Neck Surgery, Zhongnan Hospital of Wuhan University, Wuhan, China.
出版信息
Biochem Biophys Res Commun. 2022 Jul 23;614:47-55. doi: 10.1016/j.bbrc.2022.05.007. Epub 2022 May 6.
PURPOSE
Folate-mediated one-carbon metabolism (FOCM) plays a vital role in supporting cancer cells hyperproliferation. Malignant cells, including nasopharyngeal carcinoma (NPC) cells, are characterized by rapid proliferation and thus need large numbers of nucleotides and nutrients generated from FOCM. However, the mechanism and key genes involved in FOCM playing a vital role in NPC progression are still unclear. This study aimed to find out the key gene, and its functions in NPC and explore the potential mechanism.
METHODS
Bioinformatics analysis based on TCGA and GSEA database were performed to screen the key FOCM related gene in HNSCC. The effects of MTHFD2 on cell proliferation, apoptosis and migration were conducted through MTHFD2 knockdown cell lines in vitro experiments. Cell proliferation was explored by CCK8 assay and colony formation assay. Cell apoptosis was tested through flow cytometry. Transwell migration assay was performed to study the cell migration. The potential pathway was explored by RNA-seq and the ERK inhibitor SCH772984 and the ERK activator tBHQ were applied to verify the effect of MTHFD2 in NPC via the ERK pathway. Finally, xenograft tumor model was used to explore the tumorigenicity of NPC cells in vivo and IHC was performed to study the expression of related proteins.
RESULTS
MTHFD2 was highly expressed in NPC and associated with a poor prognosis. MTHFD2 knockdown inhibited the proliferation, migration and induced apoptosis of NPC cells in vitro. In consistent with cellular results, knockdown of MTHFD2 suppressed the tumorigenicity of NPC cells in vivo. MAPK pathway was enriched among DEGs between MTHFD2 knockdown cells and control cells. And the level of p-ERK1/2 and p-p38 MAPK was decreased in MTHFD2 knockdown cells and xenograft tumors of MTHFD2 knockdown cells. Furthermore, the application of the selective ERK inhibitor SCH772984 and the ERK activator tBHQ confirmed that MTHFD2-knockdown inhibited the proliferation and migration of NPC cells via the ERK signaling pathway.
CONCLUSION
MTHFD2 was up-regulated in NPC tissues and its high expression was linked to a poor prognosis. Knockdown of MTHFD2 inhibited proliferation and migration of NPC cells through the ERK signaling pathway, which may provide new clues and targets for the treatment of NPC.
目的
叶酸介导的一碳代谢（FOCM）在支持癌细胞过度增殖中起着至关重要的作用。包括鼻咽癌（NPC）细胞在内的恶性细胞的特征是快速增殖，因此需要大量来自 FOCM 的核苷酸和营养物质。然而，FOCM 在 NPC 进展中发挥重要作用的机制和关键基因仍不清楚。本研究旨在寻找关键基因及其在 NPC 中的功能，并探讨潜在的机制。
方法
基于 TCGA 和 GSEA 数据库的生物信息学分析筛选 HNSCC 中与 FOCM 相关的关键基因。通过体外实验中的 MTHFD2 敲低细胞系研究 MTHFD2 对细胞增殖、凋亡和迁移的影响。通过 CCK8 检测和集落形成实验研究细胞增殖。通过流式细胞术检测细胞凋亡。通过 Transwell 迁移实验研究细胞迁移。通过 RNA-seq 探索潜在通路，并应用 ERK 抑制剂 SCH772984 和 ERK 激活剂 tBHQ 通过 ERK 通路验证 MTHFD2 在 NPC 中的作用。最后，使用异种移植肿瘤模型在体内研究 NPC 细胞的致瘤性，并通过 IHC 研究相关蛋白的表达。
结果
MTHFD2 在 NPC 中高表达，与预后不良相关。MTHFD2 敲低抑制 NPC 细胞的增殖、迁移，并诱导细胞凋亡。与细胞结果一致，MTHFD2 敲低抑制 NPC 细胞在体内的致瘤性。MAPK 通路在 MTHFD2 敲低细胞和对照细胞之间的差异表达基因中富集。并且，MTHFD2 敲低细胞中 p-ERK1/2 和 p-p38 MAPK 的水平降低，MTHFD2 敲低细胞的异种移植肿瘤中也是如此。此外，应用选择性 ERK 抑制剂 SCH772984 和 ERK 激活剂 tBHQ 证实，MTHFD2 敲低通过 ERK 信号通路抑制 NPC 细胞的增殖和迁移。
结论
MTHFD2 在 NPC 组织中上调，其高表达与预后不良相关。MTHFD2 敲低通过 ERK 信号通路抑制 NPC 细胞的增殖和迁移，为 NPC 的治疗提供了新的线索和靶点。
Zotero 插件