天津医药

天津医药 ›› 2021, Vol. 49 ›› Issue (12): 1250-1255.doi: 10.11958/20211084

• 细胞与分子生物学 • 上一篇    下一篇

DNMT1通过下调SOX1表达参与宫颈癌生长和转移的机制研究

智艳芳，张婷，刘慧，曾宪旭，班振英，张威   

  1. 郑州大学第三附属医院病理科（邮编450052）
  • 收稿日期:2021-05-06 修回日期:2021-09-02 出版日期:2021-12-15 发布日期:2021-12-27
  • 通讯作者: 智艳芳 E-mail:zhiyanfang1980@163.com
  • 作者简介:智艳芳（1980），女，硕士，副教授，主要从事妇科肿瘤研究。E-mail：zhiyanfang1980@163.com
  • 基金资助:
    2019年度河南省医学科技攻关计划（联合共建）项目（LHGJ20190392）

Research on the mechanism of DNMT1 involved in the growth and metastasis of cervical cancer by down-regulating the expression of SOX1

ZHI Yan-fang, ZHANG Ting, LIU Hui, ZENG Xian-xu, BAN Zhen-ying, ZHANG Wei   

  1. Department of Pathology, the Third Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China
  • Received:2021-05-06 Revised:2021-09-02 Published:2021-12-15 Online:2021-12-27

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摘要： 目的 探讨DNA甲基转移酶1（DNMT1）通过调节性别决定区Y-框1（SOX1）表达对宫颈癌（CC）生长和转 移的影响。方法 采用亚硫酸氢盐定量PCR测定CC组织和细胞及CC癌旁组织和正常宫颈细胞中SOX1甲基化水 平，Western blot检测DNMT1、SOX1蛋白表达。将HeLa229和SW756细胞分为DNMT1-NC组、DNMT1-siRNA组和空 白组，采用亚硫酸氢盐定量 PCR 测定 SOX1 甲基化比例，Western blot 检测 DNMT1、SOX1、c-Myc、Cyclin D1 及 β- catenin蛋白表达，细胞克隆和裸鼠成瘤实验测定细胞增殖，Transwell实验测定细胞迁移和侵袭。结果 CC组织较癌 旁组织DNMT1表达水平及SOX1甲基化比例增高，SOX1蛋白表达降低（P＜0.05）；宫颈癌HeLa229、ME-180、SiHa、 SW756细胞较宫颈上皮Ect1/E6E7细胞DNMT1表达水平及SOX1甲基化比例增高，SOX1蛋白表达降低（P＜0.05）。 与空白组、DNMT1-NC组比较，DNMT1-siRNA组HeLa229和SW756细胞体外克隆形成数、迁移和侵袭数量减少，裸 鼠内瘤体质量减轻，DNMT1、c-Myc、Cyclin D1、核/质β-catenin蛋白表达及SOX1甲基化比例降低，SOX1蛋白表达增 加（P＜0.05）。结论 DNMT1在CC组织和细胞中高表达，可能通过维持SOX1启动区高甲基化并抑制其蛋白表达， 促进CC生长和转移。

关键词: 宫颈肿瘤, 细胞增殖, 细胞运动, 甲基转移酶类, 性别决定区Y蛋白质, 细胞侵袭, DNA甲基转移酶1, 性 别决定区Y-框1

Abstract: Objective To investigate the effect of DNA methyltransferase 1 (DNMT1) on the growth and metastasis of cervical cancer (CC) by regulating the expression of sex determining region Y-box 1 (SOX1). Methods Bisulfite quantitative PCR was used to determine the SOX1 methylation level in CC tissues and cells, CC adjacent tissues and normal cervical cells. Western blot assay was used to detect the protein expression levels of DNMT1 and SOX1. HeLa229 and SW756 cells were divided into the DNMT1-NC group, the DNMT1-siRNA group and the blank group. Bisulfite quantitative PCR was used to determine the SOX1 methylation ratio. Western blot assay was used to detect the DNMT1, SOX1, c-Myc, Cyclin D1 and β-catenin protein expression. The cell cloning and nude mouse tumor formation experiments were used to measure cell proliferation, and Transwell experiment was used to measure cell migration and invasion. Results The expression level of DNMT1 and the methylation ratio of SOX1 were higher in CC tissues than those in adjacent tissues, while the expression of SOX1 protein was significantly lower (P＜0.05). The expression level of DNMT1 and the methylation ratio of SOX1 in cervical cancer HeLa229, ME-180, SiHa, SW756 cells were significantly higher than those in cervical epithelial Ect1/E6E7 cells, while the expression of SOX1 protein was significantly lower (P＜0.05). Compared with the blank group and the DNMT1-NC group, the numbers of in vitro clone formation, migration and invasion of HeLa229 and SW756 cells decreased in the DNMT1-siRNA group, and the tumor mass was reduced in nude mice. The protein expression and SOX1 methylation ratio of DNMT1, c-Myc, Cyclin D1, and nuclear/plasma β -catenin decreased, SOX1 protein expression increased (P＜0.05). Conclusion DNMT1 is highly expressed in CC tissues and cells, and it may promote the growth and metastasis of CC by maintaining SOX1 promoter hypermethylation and inhibiting the protein expression.

Key words: uterine cervical neoplasms, cell proliferation, cell movement, methyltransferases, sex-determining region Y protein, cell invasion, DNA methyltransferase 1, sex determining region Y-box 1

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