RUVBL1表达下调对胃癌细胞生物学行为的影响及其机制  

Effects of down-expression of RUVBL1 on biological behaviors of gastric cancer cells and its mechanism

作　　者：于亭亭 徐萍 姜中华 王建华 YU Tingting;XU Ping;JIANG Zhonghua;WANG Jianhua(Department of Gastroenterology,The First People's Hospital of Yancheng,Yancheng 224006,China)

机构地区：[1]盐城市第一人民医院消化内科,江苏盐城224006

出　　处：《山东医药》2025年第1期53-58,共6页Shandong Medical Journal

基　　金：江苏省自然科学基金项目(BK20211116);江苏省盐城市基础研究计划指导性项目(YCBK202212)。

摘　　要：目的探讨RUVBL1表达下调对胃癌细胞增殖、迁移和侵袭的影响及其作用机制。方法收集胃癌组织及配对癌旁正常组织各102例,采用免疫组织化学法检测胃癌及癌旁组织RUVBL1蛋白。将人胃腺癌细胞系AGS、SGC-7901分别分为对照组及RUVBL1敲低组,对照组正常培养不进行转染,RUVBL1敲低组使用RUVBL1敲低慢病毒进行慢病毒转染。采用CCK-8实验、克隆形成实验、小鼠荷瘤实验观察各组细胞在体内外的增殖能力,采用细胞划痕实验、Transwell小室实验观察各组细胞的迁移、侵袭能力。采用转录组测序筛选各组间差异表达基因,KEGG功能富集分析差异表达基因富集的通路,发现PI3K-AKT信号通路为RUVBL1作用于胃癌细胞的关键信号通路。将AGS、SGC-7901细胞分为对照组、RUVBL1敲低组,对照组正常培养不进行处理,RUVBL1敲低组转染RUVBL1敲低慢病毒,采用Western blotting法检测细胞PI3K、AKT、p-PI3K、p-AKT蛋白。在RUVBL1敲低组的基础上加入AKT激动剂SC79作为RUVBL1敲低+AKT激动剂组,采用Western blotting法检测细胞AKT、p-AKT蛋白。采用CCK-8实验及克隆形成实验观察各组细胞增殖能力。结果胃癌组织RUVBL1蛋白表达高于癌旁组织(P<0.05)。RUVBL1敲低组在AGS、SGC-7901细胞中细胞A值、集落形成数量、荷瘤体积、荷瘤重量均低于对照组(P均<0.05),AGS、SGC-7901细胞中对照组、RUVBL1敲低组划痕愈合率及穿膜细胞数比较差异均无统计学意义(P均<0.05)。AGS、SGC-7901细胞中RUVBL1敲低组细胞p-PI3K、p-AKT蛋白表达均低于对照组,RUVBL1敲低+AKT激动剂组p-AKT蛋白表达均高于RUVBL1敲低组;RUVBL1敲低组细胞A值、集落形成数量均低于对照组、RUVBL1敲低+AKT激动剂组(P均<0.05)。结论RUVBL1在胃癌中高表达,RUVBL1表达下调可抑制胃癌细胞的增殖能力,其机制可能与激活PI3K/AKT信号通路有关。Objective To investigate the effects of down-expression of RuvB-like AAA ATPase 1(RUVBL1)on the proliferation,migration and invasion of gastric cancer cells and the mechanism of action.Methods We collected 102 cases of gastric cancer tissues and adjacent normal tissues,and then we detected the protein levels of RUVBL1 in the gastric cancer tissues and adjacent normal tissues by immunohistochemistry.AGS and SGC-7901 cells were divided into the control group and RUVBL1 knockdown group.Cells in the control group were cultured without transfection,and cells in the RUVBL1 knockdown group were transfected with RUVBL1 knockdown lentivirus.We used cell counting kit-8 assay(CCK8),colony formation assay and mouse tumor-bearing assay to observe the proliferation abilities in vivo and in vitro.We used cell Scratch assay and Transwell chamber assay to observe the migration and invasion abilities of them.We screened differentially expressed genes between groups by transcriptome sequencing,and then analyzed the pathways enriched by differentially expressed genes by KEGG functional enrichment.We found that PI3K-AKT signaling pathway were the key signaling pathway.AGS and SGC-7901 cells were divided into the control group and the RUVBL1 knockdown group.Cells in the control group were not treated,and cells in the RUVBL1 knockdown group were transfected with RUVBL1 knockdown lentivirus.Western blotting was used to detect the protein expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT),phosphorylated phosphatidylinositol 3-kinase(p-PI3K)and phosphorylated protein kinase B(p-AKT).Based on the treatment of the RUVBL1 knockdown group,the AKT agonist SC79 was added to cells,which were taken as the RUVBL1 knockdown+AKT agonist group.Western blotting was used to detect the proteins of AKT and p-AKT in cells.CCK-8 assay and colony formation assay were employed to observe the cell proliferation ability of each group.Results The expression of RUVBL1 protein in the gastric cancer tissues was higher than that in the a

关 键 词：RUVBL1蛋白 真核生物ATP酶 胃癌 细胞增殖 细胞迁移 细胞侵袭 PI3K/AKT信号通路 

分 类 号：R573[医药卫生—消化系统]