巨噬细胞游走抑制因子在乙型肝炎病毒相关性肝细胞癌中的调控作用

Regulatory role of macrophage migration inhibitory factor in hepatitis B virus-related hepatocellular carcinoma

目的探讨巨噬细胞游走抑制因子(MIF)在乙型肝炎病毒(HBV)相关性肝细胞癌(HCC)中的调控作用机制。方法通过转染lentivirus-MIF shRNA,抑制人肝癌细胞系HepG2.2.15中MIF的表达,分为转染组和空转组。应用四甲基偶氮唑蓝(MTT)法检测细胞的生长增殖变化,采用流式细胞仪观察细胞的凋亡变化,应用Western blot蛋白印记法检测MIF、核因子κB(NF-κB)和B细胞淋巴瘤/白血病-2(Bcl-2)蛋白水平变化。转染组与空转组的细胞增殖和凋亡相关实验指标采用配对资料t检验分析。结果MIF在HepG2.2.15细胞中呈高表达,抑制MIF表达后,与空转组相比,转染组HepG2.2.15细胞的活性下降(53.0±2.0)%(t=6.421、P=0.023),凋亡增加(22.5±3.0)%(t=5.837、P=0.027),同时NF-κB、Bcl-2蛋白表达分别下降(48.8±2.0)%(t=6.936、P=0.021)和(50.3±3.0)%(t=6.729、P=0.022),差异均有统计学意义。结论M IF可能通过调控NF-κB及其下游Bcl-2通路的基因表达,从而影响HepG2.2.15细胞的增殖与凋亡。抑制MIF可能会成为HBV感染相关性HCC治疗的新靶点。

Objective To investigate the regulatory mechanism of macrophage migration inhibitory factor(MIF)in hepatitis B virus(HBV)related hepatocellular carcinoma(HCC).Methods MIF expression was inhibited in human HCC cell line HepG2.2.15 by transfection of lentivirus-MIF shRNA.HepG2.2.15 cells were divided into transfection group and control group.The growth and proliferation changes of HepG2.2.15 cells were detected by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT),changes of apoptosis in the cells were observed by flow cytometry,the protein level changes of MIF,nuclear factorκB(NF-κB)and B-cell lymphoma-2(Bcl-2)were determined by Western blot.The experimental indicators of cell proliferation and apoptosis between transfection group and control group were analyzed by paired-wise data t-test.Results MIF was highly expressed in HepG2.2.15 cell.Compared with the control group,the proliferation of HepG2.2.15 cell in transfection group was decreased by(53.0±2.0)%(t=6.421,P=0.023),the apoptosis was increased by(22.5±3.0)%(t=5.837,P=0.027),while the protein expression of NF-κB and Bcl-2 were decreased by(48.8±2.0)%(t=6.936,P=0.021)and(50.3±3.0)%(t=6.729,P=0.022),respectively,all with significant differences.Conclusions MIF could affect the proliferation and apoptosis of HepG2.2.15 cells by regulating the gene expression of NF-κB and its downstream Bcl-2 pathway.Inhibition of MIF may become a novel target for treatment of HBV infection-associated HCC.