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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。
Objective: To investigate the effect of CX3CR1 on the proliferation, migration and invasion of human hepatocellular carcinoma cell line 7721 and Hep G2 and its mechanism. Methods: The gene expression of CX3CR1 (mRNA and protein) in human normal hepatocytes LO2 and two hepatocellular carcinoma cells (7721 and Hep G2) were detected by Q-PCR and Western blot respectively. The 7721 cells were transfected with plasmid expressing CX3CR1 , Transfected Hep G2 cells with CX3CR1 interfering RNA, CX3CR1 changes by Q-PCR and Western blot; MTT and flow cytometry were used to detect the proliferation of each group; colony formation assay Self-renewal and proliferation ability. The migration and invasion ability of cells in each group were detected by scratch healing and Transwell. The activation of PI3K / AKT and MAPK / ERK signal pathways were detected by Western blot. Results: The expression of CX3CR1 mRNA and protein in 7721 cells was low, while it was highly expressed in Hep G2 cells. CX3CR1 mRNA and protein levels were significantly increased in 7721 cells transfected with CX3CR1 plasmid, The expression of CX3CR1 in Hep G2 cells was significantly decreased, and the proliferation, migration and invasion were weakened. The expressions of p-AKT and p-ERK were significantly increased in transfected Hep G2 cells, ERK levels decreased. Conclusion: The chemokine CX3CR1 can promote the proliferation, migration and invasion of human hepatocellular carcinoma cells, which may be related to the activation of PI3K / AKT and MAPK / ERK signaling pathway.