吉林大学学报(医学版) ›› 2024, Vol. 50 ›› Issue (2): 371-378.doi: 10.13481/j.1671-587X.20240210

• 基础研究 • 上一篇    

沉默FOXK1基因对胃癌HGC-27细胞增殖、迁移和侵袭的影响

陈爽,李红()   

  1. 锦州医科大学基础医学院生物化学与分子生物学教研室,辽宁 锦州 121000
  • 收稿日期:2023-03-21 出版日期:2024-03-28 发布日期:2024-04-28
  • 通讯作者: 李红 E-mail:7109919@qq.com
  • 作者简介:陈 爽(1996-),女,黑龙江省安达市人,在读硕士研究生,主要从事胃癌发病机制方面的研究。
  • 基金资助:
    辽宁省教育厅面上项目(LJKZ0803)

Effect of silencing FOXK1 gene on proliferation, migration, and invasion of gastric cancer HGC-27 cells

Shuang CHEN,Hong LI()   

  1. Department of Biochemistry and Molecular Biology,School of Basic Medical Sciences,Jinzhou Medical University,Jinzhou 121000,China
  • Received:2023-03-21 Online:2024-03-28 Published:2024-04-28
  • Contact: Hong LI E-mail:7109919@qq.com

摘要:

目的 探讨沉默叉头框K1(FOXK1)对胃癌HGC-27细胞增殖、迁移和侵袭的影响,并阐明其可能的机制。 方法 基于基因表达水平值的交互式分析平台(GEPIA)数据库查询在胃癌组织和正常胃组织中FOXK1 mRNA表达水平;利用化学合成的si-FOXK1体外转染胃癌HGC-27细胞,实验分为空白对照组、nc-FOXK1组和si-FOXK1组,采用Western blotting法评估各组的转染效率,MTT法检测si-FOXK1转染后各组胃癌HGC-27细胞的增殖能力,克隆形成实验检测si-FOXK1转染后各组胃癌HGC-27细胞的克隆形成数,细胞划痕实验检测si-FOXK1转染后各组胃癌HGC-27细胞的迁移率,Transwell小室实验检测si-FOXK1转染后各组胃癌HGC-27细胞的细胞迁移数和细胞侵袭数,Western blotting法检测si-FOXK1转染后各组胃癌HGC-27细胞中核因子κB(NF-κB)通路相关蛋白[NF-κB p65和磷酸化核因子κB p65 (p-NF-κB p65)]的表达水平。 结果 GEPIA数据库查询,在胃癌组织中FOXK1 mRNA表达水平高于正常胃组织(P<0.05);Western blotting法检测,在胃癌HGC-27细胞中FOXK1蛋白表达水平高于人正常胃黏膜GES-1细胞(P<0.01);与空白对照组和nc-FOXK1组比较,si-FOXK1组FOXK1蛋白表达水平明显降低(P<0.01);MTT法检测,与空白对照组和nc-FOXK1组比较,si-FOXK1组胃癌HGC-27细胞的增殖能力降低(P<0.05);克隆形成实验检测,与空白对照组和nc-FOXK1组比较,si-FOXK1组胃癌HGC-27细胞克隆形成数减少(P<0.05);细胞划痕实验检测,与空白对照组和nc-FOXK1组比较,si-FOXK1组胃癌细胞的迁移率降低(P<0.05);Transwell小室实验检测,与空白对照组和nc-FOXK1组比较,si-FOXK1组胃癌HGC-27细胞的迁移细胞数和侵袭细胞数明显减少(P<0.05);Western blotting法检测,与空白对照组和nc-FOXK1组比较, si-FOXK1组胃癌HGC-27细胞中p-NF-κB p65蛋白表达水平降低(P<0.05)。 结论 FOXK1在胃癌HGC-27细胞中高表达,沉默FOXK1可抑制胃癌HGC-27细胞的增殖、迁移和侵袭能力,其作用机制可能与NF-κB通路有关。

关键词: 叉头框K1, 胃肿瘤, 细胞增殖, 细胞迁移, 细胞侵袭, 核因子κB

Abstract:

Objective To discuss the effect of silencing Forkhead box K1 (FOXK1) on the proliferation, migration, and invasion of the gastric cancer HGC-27 cells, and to clarify the possible mechanism. Methods The expression levels of FOXK1 mRNA in gastric cancer tissue and normal gastric tissue were consulted based on the Gene Expression Profiling Interactive Analysis (GEPIA) Database; the synthetic si-FOXK1 was used to transfect the gastric cancer HGC-27 cells in vitro, and the cells were divided into blank control group, nc-FOXK1 group, and si-FOXK1 group. Western blotting method was used to detect the transfection efficiencies of the cells in various groups; MTT assay was used to detect the proliferation abilities of the HGC-27 gastric cancer cells in various groups after transfected with si-FOXK1. The clone formation assay was used to detect the number of clone-forming of the HGC-27 gastric cancer cells after transfected with si-FOXK1;wound healing assay was used to detect the migration rates of the gastric cancer HGC-27 cells in various groups after transfected with si-FOXK1;Transwell chamber assay was used to detect the numbers of migration and invasion cells in various groups;Western blotting method was used to detect the expression levels of NF-κB pathway-related proteins [nuclear factor κB p65 (NF-κB p65) and phosphorylated NF-κB p65 (p-NF-κB p65)] in the gastric cancer HGC-27 cells in various groups after transfected with si-FOXK1. Results The GEPIA Database results showed that compared with normal gastric tissue, the expression level of FOXK1 mRNA in gastric cancer tissue was increased (P<0.05). The Western blotting method results showed that the expression level of FOXK1 protein in the gastric cancer HGC-27 cells was higher than that in normal gastric mucosal GES-1 cells (P<0.01). Compared with blank control group and nc-FOXK1 group, the expression level of FOXK1 protein in the cells in si-FOXK1 group was significantly decreased (P<0.01). The MTT assay results showed that compared with blank control group and nc-FOXK1 group,the proliferation ability of the gastric cancer HGC-27 cells in si-FOXK1 group was decreased (P<0.05). The clone formation assay results showed that compared with blank control group and nc-FOXK1 group, the number of clone-forming of the gastric cancer HGC-27 cells in si-FOXK1 group was decreased (P<0.05). The cell scratch healing results showed that compared with blank control group and nc-FOXK1 group, the migration rate of the gastric cancer HGC-27 cells in si-FOXK1 group was decreased (P<0.05).The Transwell chamber assay results showed that compared with blank control group and nc-FOXK1 group, the number of invasion cells of the gastric cancer HGC-27 cells in si-FOXK1 group was decreased (P<0.05). The Western blotting results showed that compared with blank control group and nc-FOXK1 group, the expression level of p-NF-κB p65 protein in the cells in si-FOXK1 group was decreased (P<0.05). Conclusion FOXK1 is highly expressed in the gastric cancer HGC-27 cells, and silencing FOXK1 can inhibit the proliferation, migration, and invasion abilities of the HGC-27 gastric cancer cells; its mechanism is possibly associated with the NF-κB pathway.

Key words: Forkhead box k1, Gastric neoplasm, Cell proliferation, Cell migration, Cell invasion, Nuclear factor-κB

中图分类号: 

  • R735.2