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

doi:10.13523/j.cb.20170604

中国生物工程杂志

2017, Vol. 37

Issue (6): 22-30 DOI: 10.13523/j.cb.20170604

研究报告

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

范梦恬, 陈思成, 郭杨柳, 李亚, 孙艳婷, 李汪, 施琼

重庆医科大学检验医学院临床诊断教育部重点实验室重庆 400016

Effect of CX3CR1 on Human Hepatocellular Carcinoma Cells and Its Mechanism

FAN Meng-tian, CHEN Si-cheng, GUO Yang-liu, LI Ya, SUN Yan-ting, LI Wang, SHI Qiong

Key laboratory of Clinical Laboratory Diagnostics, Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China

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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HepG2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HepG2）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HepG2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HepG2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HepG2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。

关键词： CX3CR1; 肝癌; 侵袭; 增殖; 迁移

Abstract: 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 CX3CR1 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 cells 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 MTT and clone formation assay. The migration and invasion ability were detected by Wound-healing and Transwell assays. The protein levels of PI3K/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 cell proliferation, migration and invasion abilities were decreased, with the levels of p-ERK and p-AKT down-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.

Key words: CX3CR1 Proliferation Migration Hepatocellular carcinoma Invasion

收稿日期: 2016-12-13 出版日期: 2017-06-25

ZTFLH:

Q71

基金资助: 国家自然科学基金资助项目（NSFC81672103）

通讯作者: 施琼 E-mail: anniesq8718@aliyu.com

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引用本文:

范梦恬, 陈思成, 郭杨柳, 李亚, 孙艳婷, 李汪, 施琼. 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[J]. 中国生物工程杂志, 2017, 37(6): 22-30.

FAN Meng-tian, CHEN Si-cheng, GUO Yang-liu, LI Ya, SUN Yan-ting, LI Wang, SHI Qiong. Effect of CX3CR1 on Human Hepatocellular Carcinoma Cells and Its Mechanism. China Biotechnology, 2017, 37(6): 22-30.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20170604 或 https://manu60.magtech.com.cn/biotech/CN/Y2017/V37/I6/22

[1] Moriguchi M, Umemura A.Current status and future prospects of chemotherapy for advanced hepatocellular carcinoma. Clin J Gastroenterol, 2016,9(4):184-190.
[2] Kufareva I.Chemokines and their receptors:insights from molecular modeling and crystallography. Curr Opin Pharmacol, 2016, 30:27-37.
[3] Huang F,Geng X P.Chemokines and hepatocellular carcinoma.World Journal of Gastroenterology,2010, 16(15):1832-1836.
[4] Ren H, Zhao T, Sun J.The CX3CL1/CX3CR1 reprograms glucose metabolism through HIF-1 pathway in pancreatic adenocarcinoma. J Cell Biochem,2013, 114(11):2603-2611.
[5] Yao X, Qi L, Chen X.Expression of CX3CR1 associates with cellular migration, metastasis, and prognosis in human clear cell renal cell carcinoma.Urologic Oncology,2013, 32(2):162-170.
[6] Lv C Y, Zhou T, Chen W.Preliminary study correlating CX3CL1/CX3CR1 expression with gastric carcinoma and gastric carcinomaperineural invasion. World Journal of Gastroenterology, 2014, 20(15):4428-4432.
[7] Liu J F, Tsao Y T, Hou C H.Fractalkine/CX3CL1 induced intercellular adhesion molecule-1-dependent tumor metastasis through the CX3CR1/PI3K/Akt/NF-κB pathway in human osteosarcoma. Oncotarget, 2016, Aug 12.
[8] Shao Y, Guan J Z. Advances in the treatment of primary liver.Infectious Disease Information, 2016,29(4):248-252.
[9] Brunot A, Le Sourd S. Hepatocellular carcinoma in elderly patients:challenges and solutions.J Hepatocell Carcinoma,2016, 3:9-18.
[10] Shah A D, Bouchard M J.Interstitial fluid flow increases hepatocellular carcinoma cell invasion through CXCR4/CXCL12 and MEK/ERK signaling. PLoS One,2015, 10(11):e0142337.
[11] Du D, Liu Y. The effects of the CCR6/CCL20 biological axis on the invasion and metastasis of hepatocellular carcinoma. Int J Mol Sci, 2014,15(4):6441-6452.
[12] Sadeghi M, Lahdou I.Serum levels of chemokines CCL4 and CCL5 in cirrhotic patients indicate the presence of hepatocellular carcinoma. Br J Cancer, 2015,113(5):756-762.
[13] Yeung O W, Lo C M. Alternatively activated (M2) macrophages promote tumour growth and invasiveness in hepatocellular carcinoma. J Hepatol, 2015, 62(3):607-616.
[14] Jamieson W L. CX3CR1 is expressed by prostate epithelial cells and androgens regulate the levels of CX3CL1/fractalkine in the bone marrow:potential role in prostate cancer bone tropism. Cancer Res, 2008, 68(6):1715-1722.
[15] Luo Wen, Lin Yuanlong. miRNA-296-3p modulates chemosensitivity of lung cancer cells by targeting CX3CR1. Am J Transl Res, 2016, 8(4):1848-1851.

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