摘要
目的分析肝癌患者肿瘤免疫微环境变化及相关机制。方法纳入2015年1月至2017年12月山西大医院肝细胞癌、乙型肝炎、健康志愿者各10例。检测各组外周和局部粒细胞-巨噬细胞集落刺激因子(GM-CSF)水平。流式细胞术和免疫组化检测肝癌、肿瘤边缘和正常肝组织的髓来源的抑制性细胞(MDSCs),免疫组化检测GM-CSF通路相关蛋白表达。筛选合适肝癌细胞系后,细胞转染CCR4-NOT转录复合体亚基7(CNOT7)重组质粒,随后检测信号传导及转录活化因子(STAT)等相关蛋白表达。结果肝癌组、肝炎组、对照组外周血GM-CSF水平差异无统计学意义(P>0.05)。局部GM-CSF水平,肿瘤边缘为(32.2±8.9)ng/L,高于肝癌组织的(9.7±2.7)ng/L和正常肝组织的(11.6±2.9)ng/L,差异有统计学意义(P<0.05)。肿瘤边缘MDSCs比例为(9.9±3.6)%,高于肝癌组织的(4.0±1.5)%和正常肝组织的(6.3±2.3)%,差异有统计学意义(P<0.05)。免疫组化得到相似结果。与正常肝组织比较,肝癌组织CNOT7、STAT3高表达,而STAT1低表达。筛选出HepG2人肝癌细胞进行转染。与空质粒组比较,敲除组CNOT7敲除成功,表达下调,STAT1表达增加,STAT3表达减少,GM-CSF表达明显降低(P<0.05)。结论肝细胞癌患者肿瘤边缘组织GM-CSF分泌增加,MDSCs增多。肝癌细胞HepG2敲除CNOT7,通过激活JAK/STAT信号通路,减少GM-CSF分泌。
Objective To analyze tumor immune microenvironment and related mechanisms in liver cancer.Methods We included 10 cases of hepatocellular carcinoma,hepatitis B patients and healthy volunteers from January 2015 to December 2017 in Shanxi Grand Hospital.We first detected the peripheral and local GM-CSF level in each group,detected myeloid-derived suppressor cells(MDSCs)GM-CSF and pathway-related protein expression.from liver cancer,tumor margin and normal liver tissue through flow cytometry and immunohistochemistry,Finally,we transfected the CCR4-NOT transcriptional complex subunit 7(CNOT7)recombinant plasmid in the hepatoma cell line,and then detected the related protein expression.Results There was no significant difference for peripheral blood GM-CSF level between liver cancer group,hepatitis group and control group(P>0.05).The level of local GM-CSF was(32.2±8.9)ng/L,which was higher than that of hepatocellular carcinoma(9.7±2.7)ng/L and normal liver tissue(11.6±2.9)ng/L.The difference was statistically significant(P<0.05).The proportion of MDSCs at the edge of the tumor was(9.9±3.6)% ,which was higher than that of liver cancer(4.0±1.5)% and normal liver tissue(6.3±2.3)% ,and the difference was statistically significant(P<0.05).Immunohistochemistrydata was consistent with previous data.Compared with normal liver tissue,CNOT7 and STAT3 were highly expressed in liver cancer tissues,while STAT1 was lowly expressed.HepG2 human hepatoma cells were selected for transfection.Compared with the empty plasmid group,CNOT7 expression was decreased in the knocking out group at the same time STAT1 expression was increased,STAT3 and GM-CSF expression was decreased.Conclusion In hepatocellular carcinoma,the secretion of GM-CSF increased and the number of MDSCs increased.Knocking out CNOT7 reduced GM-CSF secretion and activate the JAK/STAT signaling pathway.
作者
赵海潮
郭舜
任崇仁
任晓静
陈系东
陈昶舟
李健
贺杰峰
赵浩亮
Zhao Haichao;Guo Shun;Ren Chongren;Ren Xiaojng;Chen Xidong;Chen Changzhou;Li Jian;He Jiefeng;Zhao Haoliang(Shanxi Medical University,Taiyuan 030001,China;Department of General Surgery,Shanxi Academy of Medical Science Shanxi Dayi Hospital,Taiyuan 030032,China)
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2019年第4期259-263,共5页
Chinese Journal of Hepatobiliary Surgery
基金
山西省"136"兴医工程领军临床重点专科项目。
关键词
癌
肝细胞
免疫
细胞
基因敲除
Carcinoma,hepatocellular
Immunity,cellular
Gene knock out
作者简介
通信作者:赵浩亮,Email:haoliangzhao@hotmail.com.
