TFB1M影响线粒体质量和肝癌细胞生长与转移

TFB1M Affects Mitochondrial Mass and Hepatocellular Carcinoma Cell Growth and Metastasis

目的:前期研究发现线粒体转录因子B1(mitochondrial transcription factor B1,TFB1M)异常表达可能与肝细胞癌的恶性生长有关,但是其作用机制尚不完全清楚。方法:公共数据分析TFB1M表达水平和预后分析,R包筛选TFB1M高低表达组差异表达基因,并进行功能富集分析。小干扰片段靶向沉默TFB1M基因,qRT-PCR检测肝癌细胞中TFB1MmRNA水平,CCK-8检测细胞活力,EdU检测细胞增殖活性,流式细胞术检测细胞周期分布,Transwell小室检测细胞迁移和侵袭,Mitotracker染色线粒体,结合共聚焦显微镜逐层扫描构建细胞3D模型,计算单细胞内线粒体质量。结果:TFB1M在肝细胞癌组织中表达水平显著升高,TFB1M高表达肝细胞癌患者预后不良。功能富集结果显示,TFB1M与过氧化物酶体增殖物激活受体信号通路等有关。细胞学实验表明,沉默TFB1M后,肝癌细胞活力、增殖活性、迁移和侵袭活性均显著降低(P<0.05)。此外,沉默TFB1M后肝癌细胞周期阻滞于G1期。机制研究结果显示,沉默TFB1M后肝癌细胞线粒体质量显著增加。结论:TFB1M可能通过影响线粒体质量在肝癌细胞生长和转移过程中发挥关键作用。

Objective:The abnormally expression of mitochondrial transcription factor B1(TFB1M)might be related to the malignant growth of hepatocellular carcinoma(HCC).However,it is unknown how TFB1M regulates the growth of HCC.Methods:The expression level of TFB1M in HCC and prognosis were analyzed with public data.The differentially expressed genes in TFB1M high and low expression groups were screened by R packet,and the functional enrichment analysis was performed.Targeted silencing of the TFB1M gene was performed by small interfering fragments.TFB1M mRNA level was detected by qRT-PCR;cell viability was detected by CCK-8;cell proliferation activity was detected by EdU;cell cycle distribution was detected by flow cytometry;cell migration and invasion were detected by Transwell chamber assay.Mitochondria were stained by Mitotracker;the 3D cell model was constructed by multiple scanning of confocal microscope;and the mitochondrial mass in single cell was finally calculated.Results:The expression level of TFB1M in HCC tissue significantly increased,and high expression of TFB1M was positively correlated to poor prognosis of HCC patients.The results of functional enrichment showed that TFB1M was related to peroxisome proliferator-activated receptor signaling pathway.Cytological experiments showed that silencing TFB1M significantly decreased cell viability,proliferation activity,migration and invasion activity in HCC cells(P<0.05).In addition,silencing TFB1M arrested the cell cycle in G1 phase.The results of mechanism study showed that silencing TFB1M significantly increased the mitochondrial mass of HCC cells.Conclusion:TFB1M might play a key role in the growth and metastasis of HCC cells via regulating mitochondrial mass.