IKBKB基因敲除对胃癌细胞NLRC5、NF-κB表达以及生物学功能的影响
Effects of IKBKB gene knockdown on NLRC5 and NF-κB expression,as well as the biological function of gastric cancer cells
张刚 1高铁铭 1吴杨 1赵芝 1曾志祥 1李哲丽1
作者信息
- 1. 定州市人民医院消化内科,定州 073000
- 折叠
摘要
目的:探究 IKBKB 基因在胃癌细胞中的表达水平与 NLRC5、NF-κB 的作用关系,明确其对胃癌细胞生物学功能影响.方法:采用 pLenti-IKBKB-sgRNA 基因敲除质粒直接对人胃癌 SGC-7901 细胞系进行转染,应用免疫印迹试验(western blot,WB)检测单克隆细胞株中IKBKB 基因敲除的效果,以及 NLRC5、NF-κB 的表达水平.划痕实验和Transwell 迁移实验检测单克隆细胞的迁移能力,流式细胞术检测细胞凋亡和细胞周期.成瘤实验观察单克隆细胞对瘤体生长的影响.在临床胃癌组织和癌旁组织中采用免疫组化染色评估NLRC5、NF-κB、IKBKB 的表达状态.明确 IKBKB 与 NLRC5、NF-κB 的相关性以及临床诊断价值.结果:敲除 IKBKB 基因后人胃癌 SGC-7901 细胞中 NLRC5、NF-κB 的表达水平显著降低,细胞迁移和侵袭能力减弱.细胞发生 G1 期阻滞,降低了 SGC-79013 细胞增殖能力,细胞总凋亡率显著上升.IKBKB 敲除后 NLRC5、NF-κB 表达水平下调影响瘤体的生长发育.NLRC5、NF-κB、IKBKB 在胃癌组织中表达水平高于癌旁组织,NLRC5、NF-κB 与 IKBKB 的表达水平呈正相关.IKBKB、NLRC5、NF-κB 三者联合诊断胃癌 AUC 为 0.921(0.876～0.967).结论:IKBKB 在胃癌细胞与NLRC5、NF-κB 存在相关作用关系,协同促进胃癌的发生和发展.
Abstract
Objective To investigate the relationship between the expression level of the IKBKB gene in gastric cancer cells and the activitiesy of NLRC5 and NF-κB.The effect of IKBKB knockdown on the biological function of gastric cancer cells was clarified.Methods The human gastric cancer SGC-7901 cell line was transfected directly with the plenti-ikkbkb-sgrna gene knockdownout plasmid.Western blot(WB)was used to detect the effect of Ikkbkb gene knockout and the expression levels of NLRC5 and NF-κB in the monoclonal cell line.Scratch test and Transwell migration assay were used to assess the migratory capacity of monoclonal cells,while flow cytometry was employed to measure apoptosis and cell cycle progression.The effect of monoclonal cells on tumor growth was observed in a tumorigenesis experiment.Immunohistochemical staining was used to evaluate the expression status of NLRC5,NF-κB,and IKBKB in clinical gastric cancer tissues and adjacent tissues.To clarify their existence,expression correlation,and clinical diagnostic value.Results After knocking out the IKBKB gene,the expres-sion levels of NLRC5 and NF-κB in human gastric cancer SGC-7901 cells were significantly reduced,and the cell migration and invasion abilities were weakened.G1 phase arrest decreased the proliferation ability of SGC-79013 cells,and the total ap-optosis rate of SGC-79013 cells significantly increased.The expression levels of NLRC5 and NF-κB were downregulated after IKBKB knockout,which impacted the growth and development of tumors.The expression levels of NLRC5,NF-κB,and IKBKB in gastric cancer tissues were higher than those in adjacent tissues.Additionally,the expression levels of NLRC5 and NF-κB were positively correlated with IKBKB.The binary logistic regression model was constructed with IKBKB,NLRC5,and NF-κB as explanatory variables.The diagnostic performance of the joint model was quantified using ROC analysis,which showed an AUC of 0.921 with a 95%confidence interval of 0.876-0.967.Conclusion IKBKB has a correlation with NLRC5 and NF-κB in gastric cancer cells,and synergistically promotes the occurrence and development of gastric cancer.
关键词
B细胞κ轻肽基因增强子抑制因子/核苷酸结合结构域受体家族含CARD结构域-5/核因子κB/胃癌/细胞生物学功能/临床诊断价值Key words
inhibitor of kappa light polypeptide gene enhancer in B-cells/NLR family CARD domain containing 5/nu-clear factor kappa-B/gastric cancer/cell biological function/clinical diagnostic value引用本文复制引用
基金项目
河北省卫健委重点科技研究计划(20220432)
出版年
2024
NETL
NSTL
万方数据