ALKBH5在甲状腺癌细胞中的生物学功能

doi:10.3969/j.issn.1004-616x.2024.05.003

癌变·畸变·突变 ›› 2024, Vol. 36 ›› Issue (5): 349-358.doi: 10.3969/j.issn.1004-616x.2024.05.003

• 论著 • 上一篇

ALKBH5在甲状腺癌细胞中的生物学功能

王淑君¹, 张慧霞^1,2, 温思妮¹, 张磊³, 王鑫¹, 武安琪¹, 朱小年^1,2, 张小英^1,2, 谭盛葵⁴, 韩菲^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;
3. 桂林医学院智能医学与生物技术学院, 广西桂林 541199;
4. 右江民族医学院, 广西百色 533000

收稿日期:2023-12-26 修回日期:2024-05-08 发布日期:2024-10-15
通讯作者: 韩菲
作者简介:王淑君,E-mail:shujunwang1998@163.com。
基金资助:
2021年桂林医学院大学生创新创业训练计划(S202110601110)

Intracellular and cellular functions of ALKBH5 in thyroid cancer cells

WANG Shujun¹, ZHANG Huixia^1,2, WEN Sini¹, ZHANG Lei³, WANG Xin¹, WU Anqi¹, ZHU Xiaonian^1,2, ZHANG Xiaoying^1,2, TAN Shengkui⁴, HAN Fei^1,2

1. School of Public Health, Guilin Medical University, Guilin 541199;
2. Guangxi Key Laboratory of Environmental Exposure and Whole Life Cycle Health Technology, Guilin 541199;
3. School of Intelligent Medicine and Biotechnology, Guilin Medical University, Guilin 541199;
4. Youjiang Medical University for Nationalities, Baise 533000, Guangxi, China

Received:2023-12-26 Revised:2024-05-08 Published:2024-10-15

摘要/Abstract

摘要： 目的： 探讨N⁶-甲基腺苷(m⁶A)去甲基化酶ALKB同系物5(ALKBH5)在甲状腺癌发生发展过程中的作用及其生物学机制，为甲状腺癌的治疗提供科学依据。方法： 将ALKBH5敲低质粒转染到甲状腺乳头状癌TPC-1细胞和甲状腺未分化癌8505C细胞后，采用m⁶A甲基化定量检测试剂盒分析甲状腺癌细胞中m⁶A甲基化修饰水平；分别采用平板集落形成试验、CCK-8试剂盒、平板划痕试验、Transwell实验分析甲状腺癌细胞的增殖、迁移、侵袭能力，流式细胞术检测细胞周期分布，并通过Western blot实验检测PI3K/AKT信号通路蛋白的表达水平。结果： 与未敲低ALKBH5的细胞系相比，ALKBH5敲低后甲状腺癌TPC-1细胞和8505C细胞的m⁶A甲基化修饰水平升高，甲状腺癌细胞增殖、迁移、侵袭数目均下降，细胞周期G₂/M期阻滞(P<0.05)。Western blot实验结果显示，ALKBH5低表达时，甲状腺癌细胞PI3K/AKT信号通路中的P85、AKT、P-AKT和FAK蛋白表达水平降低，下游Cyclin通路中CDK4、CDK6、Cyclin B1、Cyclin D1、Cyclin E1蛋白表达水平亦降低，而下游P53通路中，P53蛋白表达水平升高(P<0.05)。结论： 在甲状腺癌细胞中，ALKBH5敲低后抑制甲状腺癌细胞增殖、迁移、侵袭，导致细胞周期阻滞，并通过PI3K/AKT/P53与PI3K/AKT/Cyclin轴抑制甲状腺癌的发生发展。

关键词: N⁶-甲基腺苷, ALKB同系物5, 甲状腺癌, PI3K/AKT信号通路

Abstract: OBJECTIVE： To investigate the role of N⁶-methyladenosine (m⁶A) demethylase ALKBH5 on intracellular and cellular functions of thyroid carcinoma cells in vitro. METHODS： The ALKBH5 knockdown plasmid was transfected to human thyroid papillary carcinoma cells (TPC-1) and undifferentiated thyroid carcinoma cells (8505C). Expression of m⁶A in the cancer cells were measured using a m⁶A methylation quantification detection kit. Plate cloning，CCK-8，plate scratch，and Transwell were used to analyze the proliferation，migration，and invasion ability. Flow cytometry was used to detect the cell cycle of the cancer cells and the percentage of cell in each phase. The relationship between ALKBH5 and downstream pathways was analyzed by Western blot experiments. RESULTS： Compared to cells without the ALKBH5 knockdown，the knockdown increased m⁶A expression of levels in the TPC-1 and 8505C cancer cells，and decreased the number of cell proliferation，invasion，and migration decreased (P<0.05). In addition，cells were arrested at the G₂/M phase of cell cycle. Western blot experiments showed that when the expression of ALKBH5 was low，the key genes of the PI3K/AKT signaling pathway were perturbed. Protein expressions of P85，AK，P-AKT and FAK were inhibited，as well as that of CDK4，CDK6，Cyclin B1，Cyclin D1，Cyclin E1 in the downstream Cyclin pathway，but P53 was increased (P<0.05). CONCLUSION： In the thyroid papillary carcinoma cells，ALKBH5 knockdown inhibited their proliferation，migration，invasion and cell cycle，as well as the PI3K/AKT/P53 and PI3K/AKT/Cyclin axis.

Key words: m⁶A, ALKBH5, thyroid carcinoma, PI3K/AKT signaling pathway

中图分类号:

王淑君, 张慧霞, 温思妮, 张磊, 王鑫, 武安琪, 朱小年, 张小英, 谭盛葵, 韩菲. ALKBH5在甲状腺癌细胞中的生物学功能[J]. 癌变·畸变·突变, 2024, 36(5): 349-358.

WANG Shujun, ZHANG Huixia, WEN Sini, ZHANG Lei, WANG Xin, WU Anqi, ZHU Xiaonian, ZHANG Xiaoying, TAN Shengkui, HAN Fei. Intracellular and cellular functions of ALKBH5 in thyroid cancer cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2024, 36(5): 349-358.

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ALKBH5在甲状腺癌细胞中的生物学功能

Intracellular and cellular functions of ALKBH5 in thyroid cancer cells

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