黄曲霉毒素B1对肝癌细胞HepG2生物物理学特性及细胞骨架结构的影响

Effects of aflatoxin B1 on the biophysical properties and cytoskeleton structure of hepatocellular carcinoma cell line HepG2

目的研究黄曲霉毒素B1(AFB1)对人肝癌细胞(HCCs)生物物理学特性及细胞骨架结构的影响。方法利用0、0.01、0.1、1、5、10μmol/L AFB1分别处理HepG2细胞24 h和48 h,采用CCK-8试剂盒检测细胞活力。在此基础上,分析10μmol/L AFB1对细胞渗透脆性、膜流动性、细胞电泳率和F-actin骨架结构的影响。随后,提取细胞总RNA,通过实时荧光定量PCR检测12种主要细胞骨架结合蛋白的转录水平变化。结果AFB1处理48 h时HepG2细胞的活力呈剂量依赖性增强。10μmol/L AFB1处理能够增强HepG2细胞抗低渗能力及细胞电泳率,细胞骨架F-actin含量明显增加,主要细胞骨架结合蛋白的mRNA表达发生改变。结论AFB1能够影响HepG2细胞的生物物理学特性、细胞骨架结构及其结合蛋白,这可能与其毒性作用直接相关。

Objective To investigate the effects of aflatoxin B1(AFB1)on the biophysical properties and cytoskeleton structure of human hepatocellular carcinoma cells(HCCs).Methods HepG2 cells were respectively treated with 0,0.01,0.1,1,5,10μmol/L AFB1 for 24 h and 48 h,and the cell viability was measured by CCK-8 kit.Based on this result,the influences of 10μmol/L AFB1 on the osmotic fragility,membrane fluidity,electrophoretic mobility(EPM)and F-actin structure of cells were analyzed.Subsequently,total RNAs were extracted and the transcription-level changes of twelve main cytoskeleton binding proteins were detected by real-time quantitative PCR.Results The increased viability of HepG2 cells was induced by AFB1 in a dose-dependent manner after 48 h treatment.After treated with 10μmol/L AFB1,the anti-hypotonic ability and EPM of HepG2 cells were enhanced.The content of F-actin in HepG2 cells increased obviously,while the mRNA expression levels of the main cytoskeleton binding proteins were altered.Conclusion AFB1 can affect the biophysical properties,cytoskeleton structure and its binding proteins of HepG2 cells,which may be directly related to its toxic action.