METTL14 和 lncRNA MALAT1 的 m6A 修饰在口腔鳞状细胞癌细胞增殖中的作用机制。

Mechanism of METTL14 and m6A modification of lncRNA MALAT1 in the proliferation of oral squamous cell carcinoma cells.

机构信息

Department of Gastroenterology, 923 Hospital of the Joint Logistics Support Force of the Chinese People's Liberation Army, Nanning City, Guangxi Province, China.

Cancer Biology and Immunology Laboratory, College of Dental Medicine, Columbia University Irving Medical Center, New York, New York, USA.

出版信息

Oral Dis. 2023 Jul;29(5):2012-2026. doi: 10.1111/odi.14220. Epub 2022 May 17.

DOI:10.1111/odi.14220

PMID:35467063

Abstract

OBJECTIVES

Methyltransferase-like 14 (METTL14) plays an epigenetic role in various cancer through N6-methyladenosine (m6A) modification. This study sought to analyze the mechanism of METTL14 in oral squamous cell carcinoma (OSCC) cell proliferation.

METHODS

Expression levels of METTL14, lncRNA metastasis associated with lung adenocarcinoma transcript 1 (lncRNA MALAT1), microRNA (miR)-224-5p, and histone lysine demethylase 2A (KDM2A) in OSCC tissues (N = 40), and cell lines (FaDu, SCC-25, CAL-27, and SCC-15) were detected. Cell viability and colony formation capacity were assessed. m6A level, stability, and subcellular localization of lncRNA MALAT1 were determined. Nude mouse xenograft tumor assay was performed to confirm the role of METTL14 in vivo.

RESULTS

METTL14 and lncRNA MALAT1 were upregulated, and miR-224-5p was downregulated in OSCC tissues and cells. Silencing METTL14 repressed OSCC cell viability and colony formation. Overexpression of MALAT1 and KDM2A or miR-224-5p downregulation reversed the inhibition of silencing METTL14 on OSCC cell proliferation. METTL14 induced m6A modification of MALAT1 to upregulate MALAT1. MALAT1 is comparatively bound to miR-224-5p to promote KDM2A transcription. In vivo, METTL14 promoted tumor growth via regulating MALAT1/miR-224-5p/ KDM2A.

CONCLUSIONS

Overall, our findings verified the therapeutic role of silencing METTL14 in OSCC treatment through the MALAT1/miR-224-5p/KDM2A axis.

摘要

目的

甲基转移酶样蛋白 14（METTL14）通过 N6-甲基腺苷（m6A）修饰在各种癌症中发挥表观遗传作用。本研究旨在分析 METTL14 在口腔鳞状细胞癌（OSCC）细胞增殖中的作用机制。

方法

检测 40 例 OSCC 组织和细胞系（FaDu、SCC-25、CAL-27 和 SCC-15）中 METTL14、长链非编码 RNA 肺癌转移相关转录本 1（lncRNA MALAT1）、微小 RNA（miR）-224-5p 和组蛋白赖氨酸去甲基化酶 2A（KDM2A）的表达水平。检测细胞活力和集落形成能力。测定 lncRNA MALAT1 的 m6A 水平、稳定性和亚细胞定位。进行裸鼠异种移植肿瘤实验以证实 METTL14 在体内的作用。

结果

METTL14 和 lncRNA MALAT1 在 OSCC 组织和细胞中上调，miR-224-5p 下调。沉默 METTL14 抑制 OSCC 细胞活力和集落形成。过表达 MALAT1 和 KDM2A 或下调 miR-224-5p 可逆转沉默 METTL14 对 OSCC 细胞增殖的抑制作用。METTL14 诱导 MALAT1 的 m6A 修饰以上调 MALAT1。MALAT1 与 miR-224-5p 结合以促进 KDM2A 转录。在体内，METTL14 通过调节 MALAT1/miR-224-5p/KDM2A 轴促进肿瘤生长。