上海交通大学学报（医学版）

上海交通大学学报（医学版）

• 论著（基础研究） • 上一篇    下一篇

DNMT1在胃癌组织中的表达及其对胃癌细胞增殖和迁移的影响

周燚，秦俭，李旭，朱麟，李继坤   

  1. 上海交通大学  医学院附属第一人民医院普外科， 上海 200080
  • 出版日期:2016-07-28 发布日期:2016-08-31
  • 通讯作者: 李继坤, 电子信箱: jkli65975@163.com。
  • 作者简介:周燚（1987—）， 男， 硕士生; 电子信箱: zhouyi8040@163.com。
  • 基金资助:

     国家自然科学基金（81472236）；上海市科委基金（14ZR1433400）

Expression of DNMT1 in gastric carcinoma tissue and its effects on proliferation and migration of gastric carcinoma cells

ZHOU Yi, QIN Jian, LI Xu, ZHU Lin, LI Ji-kun   

  1. Department of General Surgery， Shanghai General Hospital， Shanghai Jiao Tong University School of Medicine， Shanghai 200080， China
  • Online:2016-07-28 Published:2016-08-31
  • Supported by:

    National Natura Science Foundation of China， 81472236； Foundation of Science and Technology Commission of Shanghai Municipality， 14ZR1433400

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摘要：

目的 探讨DNA甲基转移酶1（DNMT1）对胃癌细胞增殖和迁移能力的影响及可能机制。方法 采用实时定量PCR技术检测60例胃癌患者癌组织及癌旁组织中DNMT1的mRNA水平。利用慢病毒包装建立稳定表达DNMT1-shRNA的MKN45、SGC7901、MKN28细胞系，同时设未处理的未转染组、加入慢病毒阴性对照的阴性对照组。CCK8法检测各组细胞的增殖情况，Western blotting检测CDC25B的表达水平，Transwell实验检测DNMT1对胃癌细胞迁移能力的影响。结果 52例（86.7%，52/60）患者的胃癌组织中DNMT1的mRNA水平较癌旁组织上调，其中17例（28.3%，17/60）上调2倍以上；有淋巴结转移、有癌旁血管浸润及TNM分期Ⅲ~Ⅳ期者其转录水平明显上调（P<0.05）。转染DNMT1-shRNA后3 d，MKN45、SGC7901以及MKN28细胞生长明显受抑，转染组的3株细胞吸光度值均明显低于未转染组及阴性对照组（P=0.000）。转染组CDC25B蛋白表达降低，迁移力下降（均P<0.05）。结论 DNMT1在胃癌组织中的表达明显高于癌旁组织,其具有促进胃癌细胞增殖和远处转移的作用，DNMT1可能通过调控CDC25B来发挥致癌作用。

关键词: 胃癌, 细胞增殖, 细胞迁移, DNA甲基转移酶1, CDC25B

Abstract:

Objective To explore the effects of DNA methyl transferase 1 (DNMT1) on the proliferation and migration of gastric carcinoma cells and its possible mechanisms. Methods The mRNA levels of DNMT1 in cancer tissues and adjacent normal tissues of 60 patients with gastric carcinoma were detected with qRT-PCR. Lentivirus packing was used to establish MKN45, SGC7901, and MKN28 gastric carcinoma cell lines expressing stable DNMT1-shRNA, which served as the transfection group. The untreated MKN45, SGC7901, and MKN28 cell lines served as the untransfected group and the MKN45, SGC7901, and MKN28 cell lines treated with lentivirus control served as the negative control group. Proliferation of MKN45, SGC7901, and MKN28 cells was detected by CCK8 method. The expression of CDC25B was measured by Western blotting. The effect of DNMT1 on the migration of gastric carcinoma cells was detected by cell migration assay. Results The mRNA levels of DNMT1 in 52 patients (86.7%，52/60) were up-regulated in cancer tissues than in adjacent normal tissues and the mRNA levels of DNMT1 in 17 of them (28.3%, 17/60) were up-regulated more than two times. The transcription level of DNMT1 in cancer tissues of patients with lymph node metastasis, vascular invasion, and TNM stage Ⅲ-Ⅳ was up-regulated significantly (P＜0.05). The growth of MKN45, SGC7901, and MKN28 cells was significantly inhibited 3 d after DNMT1-shRNA transfection. The absorbance values of three cell lines in the transfection group were significantly lower than those in the untransfected group and the negative control group (P=0.000). The protein expression of CDC25B and the migration ability decreased in the transfection group as compared with the untransfected group and the negative control group (both P＜0.05). Conclusion The expression of DNMT1 is significantly higher in gastric carcinoma tissues than that in adjacent normal tissues. DNMT1 can promote the proliferation and metastasis of gastric carcinoma cells, which may be associated with the regulation on CDC25B.

Key words: stomach neoplasms, cell proliferation, cell migration, DNMT1, CDC25B