肝癌细胞SMMC-7721中CXCL3表达与氧化应激的相关性研究

Correlation between CXCL3 and cellular oxidative stress in the liver cancer cell SMMC-7721

目的探讨肝癌细胞中CXCL3的表达与氧化应激水平的关系。方法生物信息学分析CXCL3在肝癌组织和正常组织中的表达差异,其表达情况与生存率的关系,并筛选与CXCL3表达相关的氧化应激基因。过氧化氢对肝癌细胞SMMC-7721进行氧化应激刺激后,qRT-PCR检测CXCL3表达变化。建立CXCL3高表达和CXCL3低表达的肝癌细胞模型,利用CCK-8实验检测细胞增殖情况,利用酶联免疫吸附试验、Western Blotting法等检测细胞中的超氧化物歧化酶(SOD)、丙二醛(MDA)、血红素氧合酶(HO-1)。结果对TCGA数据库的生物信息学分析可以发现,CXCL3在肝癌组织的转录水平与正常组织比较差异无统计学意义(t=0.864,P=0.075);CXCL3高表达患者与CXCL3低表达患者相比,总生存率较低(P=0.001);肝癌组织中有7个与CXCL3表达相关的氧化应激基因;过氧化氢刺激后肝癌细胞CXCL3表达降低(200μmol/L时t=3.275,P=0.004;400μmol/L时t=5.162,P<0.001)。高表达CXCL3的细胞增殖能力增强(t=4.180,P<0.001),低表达CXCL3的细胞增殖能力降低(t=60.201,P<0.001);高表达CXCL3的细胞SOD、MDA和HO-1升高(t=253.301,P<0.001;t=19.485,P<0.001;t=5.592,P<0.001),低表达CXCL3的细胞SOD、MDA和HO-1降低(t=253.782,P<0.001;t=11.265,P<0.001;t=13.064,P<0.001)。结论CXCL3的表达影响肝癌的恶性表型,这可能与肝癌细胞中CXCL3影响其氧化应激有关。

Objective To investigate the relationship of expression of CXCL3 with cellular oxidative stress in the liver cancer cells.Methods The expression differences of CXCL3 in liver cancer cells and normal tissues were analyzed by bioinformatics,and the relationship between its expression and survival rate,and the oxidative stress genes related to CXCL3 expression were screened.The expression of CXCL3 was detected by qRT-PCR after oxidative stress stimulation of liver cancer cell SMMC-7721 by hydrogen peroxide.Liver cancer cell models with overexpression of CXCL3 and low expression of CXCL3 were established,and the cell proliferation was detected by CCK-8 assay.Superoxide dismutase(SOD),malondialdehyde(MDA),heme oxygenase-1(HO-1)levels were detected by enzyme linked immunosorbent assay and Western blot hybridization.Results Bioinformatics analysis of the TCGA database showed that the transcription level of CXCL3 was not different between in the liver cancer tissue and the normal tissues(t=0.864,P=0.075);the overall survival rate of patients with higher CXCL3 expression levels was lower than that of patients with lower CXCL3 expression levels(P=0.001);and there were 7 oxidative stress genes associated with CXCL3 expression in liver cancer tissues,including 1 negatively correlated gene and 6 positively correlated genes;the expression of CXCL3 in hepatocellular carcinoma cells decreased after H2O2 stimulation(t=3.275,P=0.004 as 200μmol/L;t=5.162,P<0.001 as 400μmol/L).In the liver cancer cells,the proliferation ability of overexpressed cells with stable transfection of CXCL3 was stronger than that of the control group(t=4.180,P<0.001),and the proliferation ability of low-expression cells with RNAi interfering with CXCL3 was lower than that of the control group(t=60.201,P<0.001);the SOD,MDA and HO-1 levels of cells overexpressed with CXCL3 were higher than those of the control group(t=253.301,P<0.001;t=19.485,P<0.001;t=5.592,P<0.001),and the SOD,MDA and HO-1 levels of cells with low expression of CXCL3 were lower than those of the control group(t=253.782,P<0.001;t=11.265,P<0.001;t=13.064,P<0.001).Conclusion The expression level of CXCL3 affects the malignant phenotype of liver cancer;in liver cancer cells,CXCL3 is closely related to oxidative stress.