趋化因子受体CX3CR1对人肝癌细胞7721和HepG2的作用及其机制的研究

发布时间：2019-06-21 13:15

【摘要】：目的:探究趋化因子受体CX3CR1(C-X3-C motif chemokine receptor 1,CX3CR1)对人肝癌细胞7721和Hep G2增殖、迁移和侵袭的影响及其机制。方法:采用Q-PCR和Western blot法分别检测人正常肝细胞LO2和两种肝癌细胞(7721和Hep G2)中CX3CR1的基因表达情况(mRNA和蛋白质);以过表达CX3CR1的质粒转染7721细胞,用抑制CX3CR1的干扰RNA转染Hep G2细胞,通过Q-PCR和Western blot法检测CX3CR1的变化;应用MTT和流式细胞实验检测各组细胞的增殖能力;用集落形成实验检测各组细胞的自我更新和增殖能力;借助划痕愈合和Transwell检测各组细胞的迁移和侵袭能力;利用Western blot法检测PI3K/AKT、MAPK/ERK信号通路的激活情况。结果:CX3CR1在7721细胞中mRNA和蛋白质呈低表达趋势,而在Hep G2细胞中则呈高表达趋势;转染过表达CX3CR1质粒后7721细胞中CX3CR1的mRNA和蛋白水平有明显的升高,细胞的增殖、迁移、侵袭能力增强,p-AKT和p-ERK水平升高;转染干扰RNA后Hep G2细胞中的CX3CR1表达水平明显下降,增殖、迁移、侵袭能力减弱,p-AKT和p-ERK水平降低。结论:趋化因子受体CX3CR1可以促进人肝癌细胞增殖、迁移和侵袭能力,该作用可能与PI3K/AKT、MAPK/ERK信号通路激活有关。
[Abstract]:Objective: To investigate the effects and mechanisms of chemokine receptor CX3CR1 (C-X3-C motilin receptor 1, CX3CR1) on the proliferation, migration and invasion of human hepatoma cells 7721 and Hep G2. Methods: The expression of CX3CR1 (mRNA and protein) was detected by Q-PCR and Western blot respectively, and the expression of CX3CR1 in human normal hepatocytes (7721 and Hep G2) was detected by using the method of Q-PCR and Western blot. The 7721 cells were transfected with the plasmid expressing the CX3CR1, and the Hep G2 cells were transfected with the interfering RNA which inhibited the CX3CR1. The changes of CX3CR1 were detected by Q-PCR and Western blot. The proliferation ability of each group was detected by MTT and flow cytometry. The self-renewal and proliferation ability of each group of cells were detected by colony formation. The migration and invasion ability of each group of cells were detected by means of scratch-healing and Transwell. The activation of PI3K/ AKT, MAPK/ ERK signal pathway was detected by Western blot. Results: The mRNA and protein of CX3CR1 in 7721 cells showed a low expression tendency, while in the Hep G2 cells, the expression of CX3CR1 mRNA and protein in 7721 cells increased significantly, and the proliferation, migration and invasion ability of the cells increased, and the levels of p-AKT and p-ERK increased. The expression level of CX3CR1 in the Hep G2 cells after transfection of the interfering RNA was significantly decreased, the proliferation, the migration, the invasion ability decreased, the p-AKT and the p-ERK levels were reduced. Conclusion: The chemokine receptor CX3CR1 can promote the proliferation, migration and invasion of human liver cancer cells, which may be related to the activation of PI3K/ AKT, MAPK/ ERK signaling pathway.
【作者单位】：重庆医科大学检验医学院临床诊断教育部重点实验室;
【基金】：国家自然科学基金资助项目(NSFC 81672103)
【分类号】：R735.7

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