摘要
目的:探究miR-618靶向细胞因子信号转导抑制因子-1(SOCS1)对肝癌患者中树突状细胞(DC)表型及功能的调控机制。方法:收集我院58例肝癌患者外周血,分离外周血单核细胞(PBMC)后诱导分离DC。双荧光素酶报告系统以及Western blot检测用来验证miR-618与SOCS1的靶向调控关系。荧光定量PCR检测DC诱导期间miR-618表达。诱导后的DC进行miR-618过表达或SOCS1沉默处理。流式细胞术检测各组DC中CD13、CD33、CD11c、HLA-DR、CD86、CD4表面抗原。ELISA检测各组DC中TNF-γ、IL-6和IFN-α分泌水平。划痕愈合实验检测各组DC迁移能力。结果:在DC诱导期间,miR-618表达呈时间依赖性下调。双荧光素酶报告系统和Western blot结果显示miR-618能够靶向抑制SOCS1表达。与对照组相比,DC经miR-618过表达或SOCS1沉默后,CD13、CD33、CD11c、HLA-DR、和CD86阳性细胞显著增多(P<0.05),但CD4阳性细胞无显著变化(P>0.05)。此外,miR-618 mimic组和SOCS1 shRNA组DC表达TNF-γ、IL-6和IFN-α显著增强且DC迁移能力得到显著提高(P<0.05)。结论:miR-618通过靶向SOCS1来促进肝癌患者中DC活化,增强其免疫调控能力。
Objective:This study is designed to explore the underlying mechanism by which miR-618 affect the phenotype and function of dendritic cell(DC)through targeting suppressors of cytokine signaling 1(SOCS1)in patients with liver cancer.Methods:Peripheral blood of 58 cases of patients with liver carcinoma were collected,then DC were induced and isolated from peripheral blood mononuclear cells(PBMC).Dual luciferase reporter system and Western blot were utilized to confirm the targeting regulatory relations between miR-618 and SOCS1.Fluorescence quantitative PCR was used to detect miR-618 expression during induction of DC.Then induced DC were treated with miR-618 overexpression or SOCS1 silencing.Flow cytometry was adopted to measure the surface antigen CD13,CD33,CD11c,HLA-DR,CD86,CD4 in DC.ELISA was used to detect the secretion of TNF-γ,IL-6 and IFN-αin DC.Scratch healing experiment was adopted to measure the migration of DC in each group.Results:miR-618 expression dropped in time-dependent manner during the induction of DC.Results obtained from dual luciferase reporter system and Western blot revealed that miR-618 could inhibit SOCS1 expression.Compared with NC group,CD13,CD33,CD11c,HLA-DR and CD86 positive cells were significantly increased after DC were treating with miR-618 mimic or SOCS1 silencing(P<0.05),while there was no significant difference on the CD4 positive cells(P>0.05).Furthermore,TNF-γ,IL-6 and IFN-αproduction in DC increased in miR-618 mimic and SOCS1 shRNA group,at the same time,migration of DC in the above two groups was also up-regulated(P<0.05).Conclusion:miR-618 could enhance the activation of DC,improve it′s immunomodulation function in patients with liver cancer through targeting SOCS1.
作者
张壮苗
张岩
ZHANG Zhuang-Miao;ZHANG Yan(Department of Hematology Oncology,People′s Hospital of Sanya,Sanya 572000,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2020年第23期2913-2918,共6页
Chinese Journal of Immunology
