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

Effect of CX3CR1 on Human Hepatocellular Carcinoma Cells and Its Mechanism

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。

Aim：To investigate the effect of CX3CR1 on the proliferation migration and invasion abilities of hepatocellular carcinoma（HCC） cell lines and its mechanism. Methods： The expression of CX3CRI at mRNA and protein levels of 7721, HepG2 and human normal liver epithelial cells LO2 was detected by Q-PCR and Western blot. The 7721 cells were transfected with the overexpression plasmid of CX3CR1 ;the HepG2 ceils were transfected with siRNA of CX3CR1, and the expression of CX3CR1 at mRNA and protein levels were detected by Q-PCR and Western blot. The cell proliferation ability was detected by MTI＂ and clone formation assay. The migration and invasion ability were detected by Wound-healing and Transwell assays. The protein levels of PI3 K/ AKT and MAPK/ERK were detected by Western blot. Results： The expression of CX3CR1 was lower in 7721 cells than that in HepG2 cells . After overexpression plasmid was transfected into 7721 cells , the expression of CX3CR1 was up-regulated; cell proliferation migration and invasion abilities were increased. Meanwhile, the levels of p-ERK and p-AKT were up-regulated. After siRNA was thansfected into HepG2 cells , the expression of CX3CR1 was down-regulated , and levels of p-ERK and p-AKT down- cell proliferation, migration and invasion abilities were decreased, with the regulated. Conclusion ： CX3CR1 can promote the proliferation, migration and invasion abilities of hepatocellular carcinoma（HCC） cells. The activation of PI3K/AKT and MEK/ERK signaling pathway may play an important role in these processes.