【摘要】 目的:基于细胞划痕实验研究DDX20基因对胃癌细胞迁移能力的调控作用。方法:首先采用实时定量PCR检测人胃黏膜细胞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基因设计并包装过表达及干扰慢病毒,基于实时定量PCR和Western印迹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:与正常的胃黏膜细胞GES相比,DDX20基因在MGC803胃癌细胞中表达水平最高,在MKN74胃癌细胞中表达水平次之,在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,特别是sh-RNA-02干扰慢病毒后,DDX20基因及蛋白表达水平显著降低。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。结论: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。更多还原

【Abstract】 Objective: To study the effect of DDX20[DEAD(Asp-Glu-Ala-Asp) box polypeptide 20] gene on the migration of gastric carcinoma cell based on the cell wound scratch assay. Methods: The DDX20 gene expression level was assayed in gastric mucosa cell GES, gastric carcinoma cell MGC803, SGC7901 and MKN74 to determine the high or low expression cell line of DDX20 gene. The overexpression and intervention lentivirus of DDX20 gene were designed, packaged and tansfected cell lines, and the stable transfection cell lines were identified by RT-q PCR and Western blot. The cell wound scratch assay was performed and images were gathered based on the these stable transfection cell lines. Results: Compared with the gastric mucosa cell GES, the expression level of DDX20 gene in the gastric carcinoma cell of MGC803 is 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, the migration of gastric carcinoma cell was significantly accelerated when DDX20 gene was overexpressed, and inhibited when down-regulation of DDX20 gene. Conclusion: DDX20 gene is a tumor metastasis associated gene, and the migration of gastric carcinoma could be decreased as its expression inhibition, providing a potential treatment target in clinic, and exhibiting a certain application value of it in clinic.更多还原