沉默FOXK1基因对胃癌HGC-27细胞增殖、迁移和侵袭的影响
Effect of silencing FOXK1 gene on proliferation,migration,and invasion of gastric cancer HGC-27 cells
陈爽 1李红1
作者信息
- 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
- 折叠
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.
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.
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κBKey words
Forkhead box k1/Gastric neoplasm/Cell proliferation/Cell migration/Cell invasion/Nuclear factor-κB引用本文复制引用
出版年
2024
国家科技期刊平台
NSTL
维普
万方数据