微小RNA-544通过靶向RUNX3下调自然细胞毒性受体1/自然杀伤细胞蛋白46促进肝癌免疫逃逸。
MiR-544 promotes immune escape through downregulation of NCR1/NKp46 via targeting RUNX3 in liver cancer.
作者信息
Pan Chenwei, Xiang Luxia, Pan Zhenzhen, Wang Xiaodong, Li Jie, Zhuge Lu, Fang Peipei, Xie Qipeng, Hu Xuezhen
机构信息
1Department of Infectious Disease, The Second Affiliated Hospital and Yuying Children's Hospital of Wenzhou Medical University, No. 109 West College Road, Wenzhou, 325027 Zhejiang China.
2The Second School of Medicine, Wenzhou Medical University, Wenzhou, 325000 China.
出版信息
Cancer Cell Int. 2018 Apr 3;18:52. doi: 10.1186/s12935-018-0542-y. eCollection 2018.
OBJECTIVE
To study the potential role of miR-544 in the immune escape mechanism of hepatoma cells.
METHODS
Natural killer (NK) cells were collected from healthy volunteers and patients with liver cancer. Interleukin (IL)-2 activated-NK-92 cells were transfected with miR-544 inhibitor/mimic or NC/pre-NC in HepG2 co-culture system. NK-92 cells were treated with control, IL-2, IL-2 + pre-NC, IL-2 + miR-544 mimic, IL-2 + miR-544 mimic + pcDNA and IL-2 + miR-544 mimic + pcDNA-runt-related transcription factor 3 (RUNX3) groups. Mice models of liver cancer were well established. Expression of miR-544, natural cytotoxicity receptor 1 (NCR1) and RUNX3 were evaluated by quantitative real-time PCR and western blotting. Flow cytometry and ELISA were used to determine NK cell cytotoxicity and the levels of INF-γ, respectively.
RESULTS
MiR-544 was upregulated while NCR1 and RUNX3 was downregulated in NK cells of patients with liver cancer. The levels of IFN-γ and miR-544 expression were increased and decreased in IL-2 activated-NK cells, respectively. Inversely, miR-544 overexpression inhibited NK cell cytotoxicity by downregulating IFN-γ. However, miR-544 directly targeted RUNX3 and negatively regulated NCR1. Furthermore, miR-544 promoted immune escape of hepatoma cells in vivo and in vitro.
CONCLUSION
miR-544 promoted the immune escape of liver cancer cells by downregulating NCR1 via targeting RUNX3.
目的
研究miR-544在肝癌细胞免疫逃逸机制中的潜在作用。
方法
从健康志愿者和肝癌患者中收集自然杀伤（NK）细胞。在HepG2共培养系统中，将白细胞介素（IL）-2激活的NK-92细胞用miR-544抑制剂/模拟物或阴性对照（NC）/预阴性对照（pre-NC）转染。NK-92细胞分别用对照组、IL-2、IL-2 + pre-NC、IL-2 + miR-544模拟物、IL-2 + miR-544模拟物 + 质粒（pcDNA）和IL-2 + miR-544模拟物 + pcDNA- runt相关转录因子3（RUNX3）处理。建立良好的肝癌小鼠模型。通过定量实时PCR和蛋白质免疫印迹法评估miR-544、自然细胞毒性受体1（NCR1）和RUNX3的表达。分别采用流式细胞术和酶联免疫吸附测定法（ELISA）检测NK细胞的细胞毒性和干扰素-γ（INF-γ）水平。
结果
肝癌患者NK细胞中miR-544上调，而NCR1和RUNX3下调。在IL-2激活的NK细胞中，INF-γ水平升高，miR-544表达降低。相反，miR-544过表达通过下调INF-γ抑制NK细胞的细胞毒性。然而，miR-544直接靶向RUNX3并负向调节NCR1。此外，miR-544在体内和体外均促进肝癌细胞的免疫逃逸。
结论
miR-544通过靶向RUNX3下调NCR1促进肝癌细胞的免疫逃逸。
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