CNOT7基因参与诱导HepG2细胞对Vγ9Vδ2T细胞的免疫耐受  

CNOT 7 gene induces HepG2 cellular immunologic tolerance against Vγ9Vδ2T cells

作　　者：赵雷[1] 任崇仁[2] 贺杰峰[2] 赵浩亮[2] 

机构地区：[1]山西医科大学研究生院,太原030001 [2]山西医科大学附属山西大医院普外科

出　　处：《中华普通外科杂志》2017年第1期57-60,共4页Chinese Journal of General Surgery

基　　金：山西省科技攻关基金资助项目（130313021-17）

摘　　要：目的研究人CCR4-NOT转录复合体亚基7（CCR4-NOT transcription complex subunit 7 human。CNOT7）在HepG2肝母细胞癌系（hepatoblastoma G2 cell line，HepG2 cells）对Vγ9Vδ2T淋巴细胞免疫耐受中的作用及相关机制。方法用CNOT7重组质粒（recombinant plasmid of CNOT，shCNOT7）及相应对照组载体质粒转染HepG2细胞，用Vγ9Vδ2T细胞因子干预转染前后的各组细胞，流式细胞仪检测细胞凋亡水平，Westernblot方法检测CNOT7及信号传导及转录激活因子1（signal transducer and activator of transcription 1，STATl）、STAT3的蛋白表达水平。Western blot法检测HepG2肝癌细胞系及人正常肝细胞系L02中CNOT7、STAT1和STAT3蛋白的表达水平。结果V19V82T细胞因子干预后．CNOT7基因敲减后的HepG2细胞组凋亡率由（7．55％±2．63％）增加至（20．59％±3．12％）。与L02细胞组比较，HepG2细胞组中的CNOT7蛋白高表达（F=28．76，P〈0．01），STAT3蛋白高表达（F=110．29，P〈0．01），STAT1蛋白低表达（F=35．67，P〈0．01）。CNOT7基因敲除后，HepG2细胞组的STATl蛋白表达上调（t=6．69，P〈0．05）。结论CNOT7基因参与诱导HepG2细胞V19VG2T细胞的免疫耐受，敲减CNOT7基因可逆转HepG2细胞对Vγ9Vδ2T细胞的免疫耐受。Objective To study the action of CNOT 7 （ CCR4-NOT transcription complex subunit 7 human）gene and its mechanisms in the process of Vγ9Vδ2T cell immunologic tolerance of HepG2 cells （ Hepatoblastoma G2 Cell Line）. Methods The shCNOT 7 （Recombinant plasmid of CNOT 7） and control vector shRNA were transfected into HepG2 cells. Vγ9Vδ2T cytokine stimulated each group before and after cell transfection, Cell apoptosis was detected by flow cytometry （FCM）, CNOT 7 protein and STAT1, STAT3 expression level was detected by Western blot. CNOT 7, STAT1 and STAT3 protein expression levels of HepG2 liver cancer cell lines and L02 normal liver cell line was assayed by Western blot. Results When stimulated by Vγ9Vδ2T cytokine, the apoptosis rate of gene-knockdown group significantly improved from （7.55% ±2. 63% ） to （20. 59% ±3.12% ）. Compared with L02 cells,the CNOT7 protein expression of HepG2 cells increased （ F = 28.76, P 〈 0. 01 ） , STAT3 protein expression increased （ F = 110. 29, P 〈 0. 01 ） ,while STAT1 protein expression was down-regulated （ F = 35.67, P 〈 0. 01 ）. CNOT 7 knockout could induce HepG2 cells STAT1 expression （ t = 6. 69,P 〈 0. 05 ）. Conclusions CNOT 7 gene could induce HepG2 cells Vγ9Vδ2T cellular immune tolerance. CNOT 7 knockout could reverse the Vγ9Vδ2T cell immunologic tolerance of HCC .

关 键 词：癌 肝细胞 免疫耐受 基因敲除技术 

分 类 号：R735.7[医药卫生—肿瘤]