敲低Siglec-15介导STAT1通路抑制肝癌细胞SMMC-7721增殖迁移  

Knockdown of Siglec-15-mediated STAT1 pathway inhibits the proliferation and migration of SMMC-7721 in hepatocellular carcinoma cells

作　　者：关璐璐 郭彩茹 闫家伟 金自立 文学军 亓民[1] GUAN Lulu;GUO Cairu;YAN Jiawei;JIN Zili;WEN Xuejun;QI Min(Luoyang Central Hospital,Henan Luoyang 471000,China;The First People's Hospital of Xinxiang City,Henan Xinxiang 453011,China;Xinxiang Medical College,Henan Xinxiang 453000,China;University of Virginia,Charlottesville Virginia 22903,USA)

机构地区：[1]洛阳市中心医院,河南洛阳471000 [2]新乡市第一人民医院,河南新乡453011 [3]新乡医学院,河南新乡453000 [4]美国弗吉尼亚大学,弗吉尼亚州夏洛茨维尔22903

出　　处：《现代肿瘤医学》2025年第5期756-763,共8页Journal of Modern Oncology

基　　金：河南省医学科技攻关计划联合共建项目(编号:LHGJ20230818);河南省洛阳市公益性行业医疗卫生专项(编号:2302040Y)。

摘　　要：目的:探究通过敲低Siglec-15介导STAT1通路抑制肝癌细胞SMMC-7721增殖迁移。方法:首先qRT-PCR和Western blot检测正常肝细胞L02及肝癌细胞系SMMC-7721、BEL-7402及HUH7中Siglec-15的表达以及检测siRNA敲低Siglec-15的转染效率。将肝癌细胞系SMMC-7721分为NC组、si-Siglec-15组,采用MTT、细胞划痕、Transwell等实验来检测敲低Siglec-15后对肝癌细胞的增殖迁移影响。使用癌症基因组图谱数据库(TCGA),分析Siglec-15高表达和低表达组中的差异表达基因,基因组数据库(KEGG)通路富集,揭示与Siglec-15相关的信号通路。进一步通过GeneMANIA数据库预测Siglec-15基因互作及相关通路,与STAT1、STAT2、STAT3、TYROBP等相关,采用Western blot验证Siglec-15对STAT1通路的影响,最后采用Stattic(STAT抑制剂)来逆向验证其对Siglec-15的影响及对肝癌细胞的生物学作用。结果:发现Siglec-15在三种肝癌细胞系中的表达水平要明显高于L02细胞(P<0.05)以及siRNA敲低Siglec-15的转染成功,后续选取肝癌细胞SMMC-7721完成实验。SMMC-7721细胞敲低Siglec-15后,该组肝癌细胞的增殖、迁移能力均明显低于NC组(P<0.001),同时STAT1、p-STAT1、STAT3、p-STAT3蛋白的表达水平都明显降低(P<0.001);免疫共沉淀结果显示Siglec-15与STAT1蛋白互作,划痕实验结果显示用Stattic抑制STAT1通路后,抑制肝癌SMMC-7721细胞迁移(P<0.001)。结论:敲低Siglec-15介导STAT1通路抑制肝癌SMMC-7721细胞的增殖、迁移。Objective:To investigate the mechanism of knockdown of Siglec-15-mediated STAT1 pathway inhibiting the proliferation and migration of SMMC-7721 in hepatocellular carcinoma cells.Methods:Firstly,the expression of Siglec-15 in normal hepatocytes L02 and hepatocellular carcinoma cell lines SMMC-7721,BEL-7402 and HUH7 was detected by qRT-PCR and Western blot,and the transfection efficiency of siRNA knockdown of Siglec-15 was detected.The hepatocellular carcinoma cell line SMMC-7721 was divided into NC group and si-Siglec-15 group,and MTT,cell scratch and Transwell assays were used to detect the effect of knockdown of Siglec-15 on the proliferation and migration of hepatoma cells.The Cancer Genome Atlas Database(TCGA)was used to analyze the differentially expressed genes in the high and low expression groups of Siglec-15,and the Genome Database(KEGG)pathway was enriched to reveal the signaling pathways associated with Siglec-15.Furthermore,the interaction of Siglec-15 gene and related pathways was predicted by GeneMANIA database,which was related to STAT1,STAT2,STAT3,TYROBP,etc.,and the effect of Siglec-15 on STAT1 pathway was verified by Western blot,and finally Stattic(STAT inhibitor)was used to reversely verify its effect on Siglec-15 and its biological effect on hepatocellular carcinoma cells.Results:The expression level of Siglec-15 in the three hepatoma cell lines was significantly higher than that of L02 cells(P<0.05)and the transfection of Siglec-15 was successful.After knockdown of Siglec-15 in SMMC-7721 cells,the proliferation and migration ability of hepatoma cells in this group were significantly lower than those in the NC group(P<0.001),and the expression levels of STAT1,p-STAT1,STAT3 and p-STAT3 proteins were significantly decreased(P<0.001).The results of co-immunoprecipitation showed that Siglec-15 interacted with STAT1 protein,and the results of scratch assay showed that the migration of hepatocellular carcinoma SMMC-7721 cells was inhibited by inhibition of STAT1 pathway with Stattic(P<0.001).Conclus

关 键 词：Siglec-15 STAT1通路 肝癌细胞 增殖 

分 类 号：R735.7[医药卫生—肿瘤]