沉默范可尼贫血补充组Ⅰ基因对肝癌细胞增殖和周期的影响

Effect of silencing fanconi anemia complementation groupⅠon the proliferation and cycle of hepatocellular carcinoma cells

目的探究范可尼贫血补充组Ⅰ基因(FANCI)对肝细胞癌(HCC)细胞增殖和周期的影响。方法通过癌症基因组图谱(TCGA)数据库和本院临床标本,验证了FANCI表达与肝癌患者的临床相关性,并对FANCI共表达基因进行富集分析(GSEA),通过敲减FANCI基因,探索FANCI表达减少对肝癌细胞增殖和细胞周期的影响,组织样本来源于2018年10月至2019年2月收集就诊于广州市第一人民医院并被诊断为"HCC"的患者,组间比较采用t检验。结果FANCI在肝癌组织信使RNA(mRNA)(0.006±0.004比0.001±0.001,t=3.828,P<0.01)和蛋白(0.202±0.052比0.613±0.346,t=3.333,P<0.05)水平高于正常组织,高表达FANCI的HCC患者预后较差(P<0.01);单因素和多因素Cox回归分析显示FANCI是HCC预后不良的因素之一(P<0.05);GSEA富集分析显示FANCI共表达基因富集于细胞周期等生物过程中,细胞实验发现敲低FANCI后细胞增殖能力下降,细胞周期阻滞于G_(1)~S期,细胞周期蛋白D1(Cyclin D1)表达下调。结论FANCI基因在HCC中高表达,敲低FANCI后可能通过下调Cyclin D1来抑制细胞增殖。

Objective To investigate the effect of Fanconi anemia complementation groupⅠgene(FANCI)on the proliferation and cycle of hepatocellular carcinoma cells.Methods The relationship between FANCI gene expression and clinical stage and prognosis of hepatocellular carcinoma(HCC)patients was analyzed by extracting relevant clinical data and sequencing data from the Cancer Genome Atlas database(TCGA),and to perform Gene Set Enrichment Analysis(GSEA)on FANCI co-expressed genes.Exploring the effect of reduced FANCI expression on the proliferation and cell cycle of hepatocellular carcinoma cells by knocking down the FANCI gene.Tissue samples were collected from 8 patients with primary diagnosis of"HCC"in Guangzhou First People’s Hospital from October 2018 to February 2019,and t-test was used for comparison between groups.Results FANCI was upregulated at the RNA(0.006±0.004 vs.0.001±0.001,t=3.828,P<0.01)and protein levels(0.20±0.05 vs.0.61±0.34,t=3.333,P<0.05)in HCC tissues relative to normal tissues,and patients with high FANCI expression had a poorer prognosis(P<0.001);univariate(P<0.01)and multiple(P<0.05)Cox regression analyses showed that FANCI was independent prognostic factor in HCC,and GSEA enrichment analysis showed that FANCI co-expressed genes were enriched in biological processes such as cell cycle;cellular experiments revealed that knockdown of FANCI resulted in decreased cell proliferation,cell cycle arrest in G_(1)-S phase,and down-regulation of Cyclin D1 expression.Conclusion FANCI gene is highly expressed in HCC,and knockdown of FANCI may inhibit cell proliferation by downregulating Cyclin D1.