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

YU Tingting; XU Ping; JIANG Zhonghua; WANG Jianhua

doi:10.3969/j.issn.1002-266X.2025.01.012

Abstract：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 gas-tric 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 en-riched 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 RU-VBL1 knockdown lentivirus.Western blotting was used to detect the protein expression levels of phosphatidylinositol 3-ki-nase(PI3K),protein kinase B(AKT),phosphorylated phosphatidylinositol 3-kinase(p-PI3K)and phosphorylated pro-tein 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 adja-cent normal tissues(P<0.05).In AGS and SGC-7901 cells,RUVBL1 knockdown significantly reduced the cell A value,colony formation number,tumor volume and tumor weight(all P<0.05).However,there were no significant differences in the rate of wound healing or the number of transmembrane cells between the control group and the RUVBL1 knockdown group(both P>0.05).The cellular p-PI3K and p-AKT protein expression levels in AGS and SGC-7901 cells were lower in the RUVBL1 knockdown group than in the control group,and the p-AKT protein expression was higher in the RUVBL1 knockdown+AKT agonist group than in the RUVBL1 knockdown group;the cellular A-value and the number of colony for-mation were lower in the RUVBL1 knockdown group than in the control group and the RUVBL1 knockdown+AKT agonist group(all P<0.05).Conclusions The expression of RUVBL1 is high in gastric cancer.Down-regulation of RUVBL1 expression can inhibit the proliferation of gastric cancer cells,and the mechanism may be related to the activation of PI3K/AKT signaling pathway.