骨桥蛋白通过LGALS3BP调控PI3K/AKT通路促进肝癌细胞迁移OACSTPCD
Osteopontin activates the PI3K/AKT pathway by upregulating LGALS3BP in promotion of hepatoma cell migration
目的 探讨骨桥蛋白(OPN)通过半乳糖凝集素3结合蛋白(LGALS3BP)促进肝癌细胞迁移的作用及机制.方法 体外培养人肝癌细胞系SMMC-7721、稳定转染空真核表达载体的人肝癌细胞系SMMC-P及稳定转染OPN基因的人肝癌细胞系SMMC-OPN.RT-qPCR检测细胞中OPN及LGALS3BP mRNA表达情况,Western blot检测细胞中OPN、LGALS3BP和PI3K/AKT通路蛋白表达水平,细胞划痕实验检测细胞迁移能力.SMMC-OPN细胞中分别转染靶向LGALS3BP的小干扰RNA(si-LGALS3BP)及阴性对照(si-NC)后,检测细胞迁移能力变化和PI3K/AKT通路蛋白相对表达水平变化.结果 相较于亲本细胞SMMC-7721和转染空载体的细胞SMMC-P,高表达OPN的细胞SMMC-OPN迁移能力明显增强,而且细胞中LGALS3BP mRNA及蛋白表达水平显著上调,同时p-PI3K/PI3K及p-AKT/AKT蛋白相对表达量均显著升高.沉默LGALS3BP表达后,SMMC-OPN细胞的迁移能力受到明显抑制,同时p-PI3K/PI3K及p-AKT/AKT蛋白相对表达水平均显著降低.结论 OPN可以通过上调LGALS3BP的表达激活PI3K/AKT通路,从而促进肝癌细胞迁移.
Objective To investigate the effect and mechanism of osteopontin(OPN)in hepatoma cell migration through galectin-3 binding protein(LGALS3BP).Methods Human hepatoma cell lines SMMC-7721,SMMC-P(stably transfected with empty eukaryotic expression vectors),and SMMC-OPN(stably transfected with the OPN gene)were cultured.mRNA expression levels of OPN and LGALS3BP were measured by RT-qPCR.Western blot assays were used to analyze the relative protein expression of OPN and LGALS3BP and PI3K/AKT pathway.Wound healing assays were performed to explore the cell migration ability.After transfection with LGALS3BP-targeting small interfering RNA(si-LGALS3BP)or negative control small RNA(si-NC)into SMMC-OPN cells,cell migration and relative expression of PI3K/AKT pathway-related proteins were assessed.Results Compared with SMMC-7721 and SMMC-P,the migratory ability of SMMC-OPN cells was significantly reinforced,and expression of LGALS3BP was obviously upregulated at both mRNA and protein levels.Moreover,relative expression of p-PI3K/PI3K and p-AKT/AKT proteins was significantly increased.Wound healing assays showed that the si-LGALS3BP obviously suppressed the migratory ability of SMMC-OPN cells.Furthermore,relative expression of p-PI3K/PI3K and p-AKT/AKT proteins in SMMC-OPN cells was significantly decreased after transfection of si-LGALS3BP.Conclusions OPN activates the PI3K/AKT pathway by upregulating LGALS3BP expression to promote hepatoma cell migration.
邓林林;安日问;赵方新;林婷;刘翠华;红梅;武建强;张烜
内蒙古医科大学基础医学院,呼和浩特 010059
骨桥蛋白LGALS3BPPI3K/AKT通路肝癌细胞迁移
osteopontinLGALS3BPPI3K/AKT pathwayliver cancercell migration
《中国比较医学杂志》 2024 (002)
9-15 / 7
国家自然科学基金(81660482);内蒙古自治区自然科学基金(2022MS08009);内蒙古自治区卫生健康科技计划项目(202202128).
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