生物技术进展 ›› 2017, Vol. 7 ›› Issue (4): 331-337.DOI: 10.19586/j.2095-2341.2017.0018

• 研究论文 • 上一篇    下一篇

DDX20基因对胃癌细胞迁移能力的调控作用

刘骏1,2,周春艳2,金丽2,陈钢2,肖倩2,3*,朱焕章1*   

  1. 1.复旦大学生命科学院, 上海 200438;
    2.上海复百澳生物科技有限公司, 上海 201210;
    3.耶鲁大学医学院, 美国, 康涅狄格州 纽黑文 06520
  • 收稿日期:2017-03-21 出版日期:2017-07-25 发布日期:2017-05-02
  • 通讯作者: 肖倩，助理研究员，研究方向为细胞生物学。E-mail: qian.xiao@yale.edu;朱焕章，教授，博士生导师，研究方向为分子细胞生物学。E-mail: hzzhu@fudan.edu.cn
  • 作者简介:刘骏，硕士研究生，研究方向为分子细胞生物学。E-mail: liujun@fubio.cn。
  • 基金资助:
    中国博士后科学基金(2013M540396)资助。

Regulation of DDX20 Gene on the Migration of Gastric Carcinoma Cell

LIU Jun, ZHOU Chunyan, JIN Li, CHEN Gang, XIAO Qian, ZHU Huanzhang   

  1. 1.School of Life Science, Fudan University, Shanghai 200438, China；
    2.Fubio Biological Technology Co. Ltd., Shanghai 201210, China；
    3.School of Medicine, Yale University, State of Connecticut New Haven, the United States
  • Received:2017-03-21 Online:2017-07-25 Published:2017-05-02

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摘要： 基于细胞划痕实验研究了DDX20基因对胃癌细胞迁移能力的调控作用。首先采用实时定量PCR检测人胃粘膜细胞GES、胃癌细胞MGC803、SGC7901和MKN74中DDX20基因的表达水平，确定DDX20基因高表达和低表达细胞株；进而针对DDX20基因设计、包被过表达并干扰慢病毒，基于实时定量PCR和Western Blot筛选稳定转染细胞株；基于稳定转染细胞株进行细胞划痕实验并采集图片。结果显示，与正常的胃粘膜细胞GES相比，DDX20基因在MGC803胃癌细胞中表达水平最高、MKN74胃癌细胞中表达水平次之、SGC7901胃癌细胞中表达水平最低。转染DDX20过表达慢病毒后，DDX20基因及蛋白表达水平显著增加；转染DDX20干扰慢病毒后，DDX20基因及蛋白表达水平显著降低，特别是转染干扰组2干扰慢病毒。过表达DDX20基因能显著促进胃癌细胞的迁移，抑制DDX20基因的表达可显著降低胃癌细胞的迁移。结果表明，DDX20基因是一个肿瘤转移相关基因，通过抑制该基因的表达能有效降低肿瘤细胞的迁移，为胃癌的临床治疗提供了一个潜在靶点，具有一定的临床应用价值。

关键词: 胃癌细胞, DDX20, 稳定转染细胞株, 细胞迁移

Abstract: To study the regulation of DDX20 gene on the migration of gastric carcinoma cell based on the cell wound scratch assay, the DDX20 gene expression level assay of gastric mucosa cell GES, gastric carcinoma cell MGC803, SGC7901 and MKN74 were further detected by real-time PCR to confirm the high and low expression cell line of DDX20 gene. We designed and packaged DDX20 gene, and interfered Lentivirus. Then we screened the stable transfection cell lines by the assay of real-time quantitive polymerase chain reaction (RT-qPCR) and Western blot, and finally perform the cell wound scratch assay and gathered images based on the stable transfection cell lines. Results showed that, compared with the gastric mucosa cell GES, the expression level of DDX20 gene in the gastric carcinoma cell of MGC803 was the highest, and second in the gastric carcinoma cell of MKN74, and the minimum in the gastric carcinoma cell of SGC7901. After transfection of the DDX20 gene overexpression Lentivirus, the expression level of DDX20 gene and protein was significantly increased. After transfection of DDX20 gene intervention Lentivirus, the expression level of DDX20 gene and protein was significantly decreased, especially in the sh-RNA-02 intervention Lentivirus. In addition, overexpressed DDX20 gene could significantly accelerate the migration of gastric carcinoma cell,  inhibited  down-regulated the expression of DDX20 gene  could inhibit the migration of gastric carcinoma cell. DDX20 gene is a tumor metastasis associated gene, and the migration of gastric carcinoma could be decreased when the expression of DDX20 was inhibited, and the results provided a potential treatment target in clinic, and exhibited a certain application value of DDX20 in clinic.

Key words: gastric carcinoma cell, DDX20, stable transfection cell lines, cell migration

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