METTL3通过m6A甲基化修饰影响葡萄膜黑色素瘤细胞增殖、迁移和侵袭OA北大核心
METTL3 Regulates the Proliferation,Migration,and Invasion of Uveal Melanoma Cell via Modifying m6A Methylation
[目的]探讨METTL3对葡萄膜黑色素瘤细胞的恶性生物学行为影响,并验证其是否和N6-甲基腺苷(m6A)甲基化相关.[方法]用慢病毒转染构建敲低、过表达METTL3以及点突变m6A相关催化位点的葡萄膜黑色素瘤细胞模型,通过Transwell了解细胞迁移、侵袭情况,利用CCK-8检测细胞增殖能力,流式细胞仪分析细胞凋亡及周期的变化.[结果]敲低METTL3的C918和MUM-2B细胞在增殖、迁移和侵袭能力方面明显下降(P<0.001),细胞凋亡率增加,两种细胞中G1期的比例显著上升,而S期细胞的比例显著下降;过表达METTL3的C918和MUM-2B细胞在增殖、迁移和侵袭能力方面明显增强(P<0.001),细胞凋亡率减少;C918细胞中处于G1期的细胞比例显著下降,而S期细胞的比例显著上升;MUM-2B细胞周期的比例没有明显变化;点突变m6A相关催化位点后细胞的增殖、迁移和侵袭能力下降,细胞凋亡率增加;MUM-2B细胞中G1期细胞的百分比显著提高,S期细胞的百分比显著下降,G2期细胞的百分比稍下降;C918细胞中G1期细胞的百分比显著提高,S期和G2期细胞比例无明显变化.[结论]葡萄膜黑色素瘤细胞的增殖、侵袭和转移能力与METTL3的表达呈正相关,细胞凋亡率与METTL3的表达呈负相关;METTL3含量的变化对不同细胞的周期干扰不一;METTL3对葡萄膜黑色素瘤细胞的增殖、迁移和侵袭的相关影响和m6A甲基化修饰相关.
[Objectives]To investigate the effect of METTL3 on the malignant biological behaviors of uveal melanoma cells and to verify whether this effect is related to m6A methylation.[Methods]Uveal melanoma cell models with METTL3 knockdown,overexpression,and point mutations at m6A-related catalytic sites were constructed via lentivirus transfection.Transwell assays were used to assess cell migration and invasion;CCK8 assays were used to measure cell proliferation and flow cytometry was used to analyze apoptosis and cycle changes.[Results]The proliferation,migration and invasion abilities of C918 and MUM-2B cells with METTL3 knockdown were significantly decreased(P<0.001),while the apoptosis rate was increased.The proportion of cells in G1 phase significantly increased,whereas the proportion in the S phase significantly decreased.Cells overexpressing METTL3 showed significantly enhanced proliferation,migration and invasion abilities(P<0.001),along with a decreased apoptosis rate.In C918 cells,the proportion of cells in G1 phase decreased significantly,while the proportion in S phase increased significantly.The cell cycle distribution of MUM-2B cells did not change remarkably.Following point mutation of m6A-related catalytic sites,cell proliferation,migration and invasion decreased,and the apoptosis rate increased.In MUM-2B cells,the percentage of cells in G1 phase significantly increased;the percentage in S phase significantly decreased and the percentage in G2 phase slightly decreased.In C918 cells,the percentage of G1 phase cells significantly increased,with no significant changes in the proportions of S and G2 phases.[Conclusions]The proliferation,invasion and metastasis of uveal melanoma cells were positively correlated with the expression of METTL3,while the apoptosis rate was negatively correlated.Changes in METTL3 levels differentially affect the cell cycles in different cell lines.The effects of METTL3 on the proliferation,migration and invasion of uveal melanoma cells are related to m6A methylation modification.
王美岚;赵天齐;师齐;郝莉莉;刘莲
暨南大学附属第一医院眼科,广东 广州 510630暨南大学附属第一医院眼科,广东 广州 510630暨南大学附属第一医院眼科,广东 广州 510630暨南大学附属第一医院眼科,广东 广州 510630暨南大学附属第一医院眼科,广东 广州 510630
医药卫生
葡萄膜黑色素瘤甲基转移酶样蛋白3N6-甲基腺苷增殖侵袭
uveal melanomamethyltransferase-like protein 3N6-methyladenosineproliferationinvasion
《中山大学学报（医学科学版）》 2025 (4)
628-638,11
广州市科技计划项目(202201020015,2025A04J3477)
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