[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(Hepato-cellular 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.
Abstract
[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 Es-timation Resource(TIMER),TNMplot and GEPIA2(Gene Expression Omnibus)and other public databases were used to ana-lyze 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 significant-ly increased compared with normal hepatocellular carcinoma cells L02(P＜0.01).The results of MTT and cell clonal forma-tion 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.
维普
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