m6A去甲基化酶FTO在甲状腺癌细胞中的作用及机制研究

doi:10.3969/j.issn.1004-616x.2023.04.002

癌变·畸变·突变 ›› 2023, Vol. 35 ›› Issue (4): 253-260.doi: 10.3969/j.issn.1004-616x.2023.04.002

m⁶A去甲基化酶FTO在甲状腺癌细胞中的作用及机制研究

田润¹, 温思妮¹, 刘翰元¹, 韩菲^1,2, 朱小年^1,2, 谭盛葵^1,2, 张慧霞^1,2

1. 桂林医学院公共卫生学院, 广西桂林 541199;
2. 广西环境暴露与全生命周期健康科技重点实验室, 广西桂林 541199

收稿日期:2023-03-26 修回日期:2023-05-15 出版日期:2023-07-30 发布日期:2023-08-04
通讯作者: 张慧霞
作者简介:田润，E-mail：tianrun@stu.scu.edu.cn。
基金资助:
2021年大学生创新创业训练计划项目(S202110601110)；2020年桂林医学院硕士研究生科研项目(GYYK2021005)；2020年联合培养基地研究生教育创新计划项目桂林医学院硕士研究生科研项目(GWXY202002，GWXY202010)

Involvement of m⁶A demethylase FTO in thyroid cancer cells

TIAN Run¹, WEN Sini¹, LIU Hanyuan¹, HAN Fei^1,2, ZHU Xiaonian^1,2, TAN Shengkui^1,2, ZHANG Huixia^1,2

1. School of Public Health, Guilin Medical College, Guilin 541199;
2. Guangxi Key Laboratory of Environmental Exposure and Life Cycle Health Science and Technology, Guilin 541199, Guangxi, China

Received:2023-03-26 Revised:2023-05-15 Online:2023-07-30 Published:2023-08-04

摘要/Abstract

摘要： 目的：探讨N⁶-甲基腺苷(m⁶A)去甲基化酶脂肪质量和肥胖相关蛋白(FTO)对甲状腺癌细胞m⁶A甲基化和增殖、转移、侵袭等生物学行为的影响，及其对MAPK-JNK信号通路的调控机制，为甲状腺癌的防治提供理论基础。方法：使用FTO敲低质粒分别转染人甲状腺乳头状癌细胞TPC-1、人甲状腺鳞癌细胞SW-579、人甲状腺未分化癌细胞8505C后，经m⁶A甲基化定量检测试剂盒检测甲状腺癌细胞m⁶A甲基化修饰水平；分别采用平板集落形成实验、CCK-8实验，划痕实验，Transwell实验检测甲状腺癌细胞的增殖、迁移及侵袭能力；Western blot法检测MAPK-JNK信号通路相关蛋白表达水平的变化。结果：与未敲低FTO的细胞系比较，FTO敲低可提高甲状腺癌TPC-1、SW-579和8505C细胞的m⁶A甲基化修饰水平，并促进甲状腺癌细胞的增殖、侵袭和迁移能力(P<0.01)。Western blot实验结果显示，FTO敲低后甲状腺癌细胞中E-cadherin和β-catenin的表达水平降低，N-cadherin和vimentin的表达升高，MAPK信号通路中的C-JUN和MLK3表达明显上调(均为P<0.05)。结论：在甲状腺癌细胞中，FTO敲低后促进甲状腺癌细胞的增殖、迁移和侵袭能力，并激活了MAPK信号通路，从而影响甲状腺癌的发生发展。

关键词: 甲状腺癌, N⁶-甲基腺苷, 去甲基化酶, 脂肪质量和肥胖相关基因, MAPK信号通路

Abstract: OBJECTIVE： To explore the effect of m⁶A demethylase FTO on m⁶A methylation，proliferation，metastasis，invasion and other biological behaviors of thyroid cancer cells，and its involvement with the MAPK-JNK signaling pathway. METHODS： The FTO knockdown plasmid was transfected into human thyroid papillary carcinoma cells (TPC-1)，thyroid squamous carcinoma cells (SW-579)，and undifferentiated thyroid carcinoma cells (8505C). Expression of m⁶A in the cancer cells were measured using a m⁶A methylation quantification detection kit. Their proliferation，migration，and invasion abilities were evaluated using the colony formation，CCK-8，wound healing，and Transwell assays. Changes in expression levels of proteins in the MAPK-JNK signaling pathway were examined using Western blot analysis. RESULTS： FTO knockdown increased m⁶A expression levels in the TPC-1，SW-579，and 8505C cancer cells，and promoted their proliferation，invasion，and migration abilities (compared to cells without FTO knockdown，P<0.01). Western blot analysis revealed that FTO knockdown led to decreased expression of E-cadherin and β-catenin，increased expression of N-cadherin and vimentin，and upregulated expression of C-JUN and MLK3 in the MAPK signaling pathway (compared with cells without FTO knockdown，all P<0.05). CONCLUSION： FTO knockdown enhanced proliferation，migration，and invasion abilities of thyroid cancer cells in vitro，and activated the MAPK signaling pathway，thereby potentially influenced development of thyroid cancer. These findings provide insights into the role of FTO demethylase in m⁶A methylation and in thyroid cancer progression.

Key words: thyroid cancer, m⁶A, demethylase, FTO, MAPK signaling pathway

中图分类号:

田润, 温思妮, 刘翰元, 韩菲, 朱小年, 谭盛葵, 张慧霞. m⁶A去甲基化酶FTO在甲状腺癌细胞中的作用及机制研究[J]. 癌变·畸变·突变, 2023, 35(4): 253-260.

TIAN Run, WEN Sini, LIU Hanyuan, HAN Fei, ZHU Xiaonian, TAN Shengkui, ZHANG Huixia. Involvement of m⁶A demethylase FTO in thyroid cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2023, 35(4): 253-260.

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m⁶A去甲基化酶FTO在甲状腺癌细胞中的作用及机制研究

Involvement of m⁶A demethylase FTO in thyroid cancer cells

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