摘要
目的探讨白细胞介素-8(IL-8)在肝癌细胞中的表达及其对增殖、迁移、侵袭的影响并分析其可能作用机制。方法采用qRT-PCR与Western blot方法分别检测肝癌细胞系HepG2、SMMC-7721及正常肝细胞L02中IL-8 mRNA和蛋白表达水平。将IL-8 siRNA转染至HepG2细胞,MTT检测沉默IL-8对HepG2细胞增殖能力的影响,Transwell迁移及侵袭实验检测沉默IL-8对HepG2细胞迁移及侵袭能力的影响,采用qRT-PCR与Western blot方法分别检测MMP-2、MMP-9 mRNA和蛋白表达水平。Western blot法检测沉默IL-8对磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(AKT)信号通路蛋白表达的影响。HepG2细胞中加入PI3K/Akt信号通路抑制剂(LY294002),观察其对HepG2细胞增殖、迁移及侵袭的影响。结果相较于L02相,肝癌细胞系HepG2、SMMC-7721中IL-8 mRNA及蛋白表达均显著升高(P<0.05);HepG2细胞中敲低IL-8后细胞增殖、迁移及侵袭能力均明显减弱,MMP-2、MMP-9及PI3K、p-AKT表达水平均明显降低;加入LY294002可明显抑制HepG2细胞增殖、迁移及侵袭能力。结论IL-8可能通过激活PI3K/Akt信号通路进而增强肝癌细胞增殖、迁移、侵袭能力。
Objective To investigate the expression of interleukin-8(IL-8)in hepatocarcinoma cells and its effects on proliferation,migration and invasion,and to analyze its possible mechanism.Methods The expressions of IL-8 mRNA and protein in hepatocellular carcinoma cell lines HepG2,SMMC-7721 and normal liver cells were detected by qRT-PCR and Western blot.IL-8 siRNA was transfected into HepG2 cells,MTT assay was used to detect the effect of IL-8 on HepG2 cell proliferation,Transwell migration and invasion assay were used to detect the effect of silencing IL-8 on HepG2 cell migration and invasion.The expression levels of MMP-2 and MMP-9 mRNA and protein were detected by qRT-PCR and Western blot,respectively.The effect of silencing IL-8 on the expression of phosphatidylinositol 3-kinase(PI3 K)/protein kinase B(AKT)signaling pathway protein was detected by Western blot.PI3 K/Akt signaling pathway inhibitor(LY294002)was added to HepG2 cells to observe its effects on proliferation,migration and invasion of HepG2 cells.Results Compared with L02 phase,IL-8 mRNA and protein expression in HepG2 and SMMC-7721 cells were significantly increased(P<0.05).The proliferation,migration and invasion of cells in HepG2 cells were significantly decreased after knocking down IL-8,and the expression levels of MMP-2,MMP-9,PI3 K and p-AKT were significantly decreased.The addition of LY294002 significantly inhibited the proliferation,migration and invasion of HepG2 cells.Conclusion IL-8 may enhance the proliferation,migration and invasion of hepatoma cells by activating PI3 K/Akt signaling pathway.
作者
余亮
江惠丽
刘豪杰
YU Liang;JIANG Hui-li;LIU Hao-jie(Department of Gastroenterology,Central Hospital of Ezhou,Ezhon 436000,China)
出处
《现代消化及介入诊疗》
2020年第2期163-168,共6页
Modern Interventional Diagnosis and Treatment in Gastroenterology
基金
国家自然青年科学基金项目(81602117)
