FTH1激活RAF/MEK/ERK通路促进肝细胞癌生长

FTH1 promotes hepatocellular carcinoma growth via activating RAF/MEK/ERK pathway

[目的]应用生物信息学和分子生物学手段研究铁蛋白重链1(Ferritin heavy chain 1,FTH1)在肝细胞癌(Hepatocellular carcinoma, HCC)中的功能和分子机制。[方法]通过TCGA(The Cancer Genome Atlas)、TIMER(Tumor Immune Estimation Resource)、TNMplot和GEPIA2(Gene Expression Omnibus)等公开数据库的数据进行分析FTH1在肝癌中的表达量和预后生存期,同时应用分子生物学实验在肝癌细胞系中分析FTH1的作用机制。[结果]在肝癌组织中FTH1的表达量高于正常(P<0.01),同时FTH1高表达的HCC患者预后更差(P<0.01)。蛋白免疫印迹分析和荧光定量PCR细胞实验表明,与正常肝细胞L02相比,肝癌细胞系HepG2和Huh7中FTH1的mRNA及蛋白水平明显升高(P<0.01)。MTT和细胞克隆形成实验结果表明FTH1敲低的HepG2和Huh7细胞增殖能力降低40%~50%。进一步蛋白免疫印迹结果显示FTH1敲低的细胞中RAF、MEK和ERK的磷酸化水平明显降低(P<0.05)。[结论] FTH1可通过激活RAF/MEK/ERK信号通路促进HCC细胞的生长。

[Objective] By bioinformatics and molecular biology methods, the function and mechanism of ferritin heavy chain 1(FTH1) in hepatocellular carcinoma(HCC) were studied.[Method] The Cancer Genome Atlas(TCGA),Tumor Immune Estimation Resource(TIMER),TNMplot and GEPIA2(Gene Expression Omnibus) and other public databases were used to analyze the expression level and prognosis survival of FTH1 in HCC,and molecular biology experiments were used to analyze the mechanism of FTH1 in HCC cells.[Result] The expression of FTH1 in HCC tissues was higher than normal(P<0.01),and HCC patients with high expression of FTH1 had worse prognosis(P<0.01).Western Blotting analysis and quantitative PCR cell assay showed that FTH1 mRNA and protein levels in hepatocellular carcinoma cell lines HepG2 and Huh7 were significantly increased compared with normal hepatocellular carcinoma cells L02(P<0.01).The results of MTT and cell clonal formation experiments showed that the proliferation capacity of HepG2 and Huh7 cells with FTH1 knockdown was reduced by 40%-50%.Further Western Blotting results showed that the phosphorylation levels of RAF,MEK and ERK were significantly reduced in HCC cells with FTH1 knockdown(P<0.05).[Conclusion] FTH1 could promote the growth of HCC cells by activating the RAF/MEK/ERK signaling pathway.