纤维蛋白原相关蛋白1-淋巴细胞活化基因3与长链非编码RNA-T细胞免疫球蛋白黏蛋白3共调控增强CD8^(+)T细胞抑制肝细胞癌肿瘤生长  

Co-regulation of fibrinogen-like protein 1-lymphocyte activation gene 3 and long non-coding RNA-T cell immunoglobulin mucin 3 enhances CD8^(+)T cell activity to suppress tumor growth in hepatocellular carcinoma

作　　者：阮玉华 杨念印[1] 卓晗 朱德明 季劼[2] Ruan Yuhua;Yang Nianyin;Zhuo Han;Zhu Deming;Ji Jie(Department of Infectious Management and Perioperative Center,Fourth Affiliated Hospital of Nanjing Medical University,Nanjing 210031,China;Department of General Surgery,the First Affiliated Hospital of Nanjing Medical University,Nanjing 210029,China)

机构地区：[1]南京医科大学第四附属医院感染管理科、围手术中心,210031 [2]南京医科大学第一附属医院普外科,210029

出　　处：《中华实验外科杂志》2022年第10期1844-1847,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(81902925)。

摘　　要：目的体外研究协同抑制纤维蛋白原相关蛋白1(FGL1)-淋巴细胞活化基因3(LAG-3)与长链非编码RNA(lncRNA)-T细胞免疫球蛋白黏蛋白3(Tim3)双免疫逃逸通路,联合增强T淋巴细胞活性,显著抑制HCC肿瘤细胞恶性增殖和肿瘤生长的效用及机制。方法构建敲除、稳定转染的5组人肝癌细胞株HepG2、HepG2-FGL1-KO、HepG2-FGL1-KO-Tim3(-)、HepG2-LAG-3-KO、HepG2-LAG-3-KO-Tim3(-),并以此构建5组细胞株移植瘤免疫系统人源化小鼠成瘤模型。通过质量细胞计数法测定淋巴细胞CD8+T亚群表达水平,并测定肿瘤生长大小体积变化。采用t和χ^(2)检验。结果实验期间,5组中位肿瘤生长速率分别为149、136、81、75、68 mm^(3)/d。5组中位肿瘤体积分别为1.18、0.91、0.37、0.45、0.29 g。与HepG2组比较HepG2-FGL1-KO组和HepG2-LAG-3-KO组动物瘤体积显著小于HepG2组(t=7.582,P<0.05),HepG2-FGL1-KO-Tim3(-)和HepG2-LAG-3-KO-Tim3(-)组动物肿瘤体积又显著小于单独敲除组(t=3.953,P<0.05)。同时,5组中位CD8+T细胞亚群表达量分别为2.1×107、2.9×107、3.8×107、7.7×107、13.2×107。HepG2-FGL1-KO-Tim3(-)和HepG2-LAG-3-KO-Tim3(-)组CD8+T细胞亚群表达活性显著高于FGL1和LAG-3单抑制组(t=2.988,P<0.05),而两个单抑制组的CD8+T细胞亚群表达活性又显著高于HepG2对照组(t=6.416,P<0.05)。结论单一免疫检查点的抑制对于遏制肿瘤逃避免疫杀伤存在局限性,多个免疫检查点因子信号通路联合抑制相较于单一通路的单控,将显著调控细胞毒性T淋巴细胞(CTL)的激活和对肿瘤细胞的敏感性毒性表达。Objective To study the effect and mechanism of synergistically inhibiting fibrinogen-like protein 1(FGL1)-lymphocyte-activation gene 3(LAG-3)and lncRNA-T cell immunoglobulin mucin 3(Tim3)dual immune escape pathways in vitro,and jointly enhancing the activity of T lymphocytes,significantly inhibiting the malignant proliferation and tumor growth of hepatocellular carcinoma(HCC)tumor cells.Methods We constructed five groups of knockout and stably transfected human hepatoma cell lines,HepG2,HepG2-FGL1-KO,HepG2-FGL1-KO-Tim3(-),HepG2-LAG-3-KO,HepG2-LAG-3-KO-Tim3(-),and constructed 5 groups of cell line transplanted tumor immune system humanized mouse tumorigenic models.The expression levels of lymphocyte CD8+T subsets were determined by mass cytometry,and the changes in tumor growth size and volume were determined.Using t andχ^(2) inspection.Results During the experiment,the median tumor growth rates of the five groups were 149,136,81,75 and 68 mm^(3)/d.The median tumor volumes of the five groups were 1.18,0.91,0.37,0.45 and 0.29 g.Compared with HepG2 group,the tumor volume of HepG2-FGL1-KO group and HepG2-LAG-3-KO group was significantly smaller than that of HepG2 group(t=7.582,P<0.05),while HepG2-FGL1-KO-Tim3(-)and HepG2-LAG-3-KO-Tim3(-)group animal tumor volume was significantly smaller than the single knockout group(t=3.953,P<0.05).At the same time,the median expression levels of CD8+T cell subsets in the five groups were 2.1×107,2.9×107,3.8×107,7.7×107 and 13.2×107.The expression activity of CD8+T cell subsets in the HepG2-FGL1-KO-Tim3(-)and HepG2-LAG-3-KO-Tim3(-)groups was significantly higher than that in the FGL1 and LAG-3 single inhibition groups(t=2.988,P<0.05),while the expression activity of CD8+T cell subsets of the two single inhibition groups was significantly higher than that of HepG2 control group(t=6.416,P<0.05).Conclusion The research results suggest that the inhibition of a single immune checkpoint is limited in preventing tumors from evading immune killing,and fully highlights that the combin

关 键 词：纤维蛋白原相关蛋白1-淋巴细胞活化基因3 长链非编码RNA-T细胞免疫球蛋白黏蛋白3 协同效应 免疫逃逸 T淋巴细胞表达 肝细胞癌 肿瘤生长 

分 类 号：R735.7[医药卫生—肿瘤]