CNOT7基因参与诱导HepG2细胞对Vγ9Vδ2T细胞的免疫耐受
发布时间：2018-10-22 10:40
【摘要】：目的:研究人CCR4-NOT转录复合体亚基7(CCR4-NOT transcription complex subunit7 human,CNOT7)在Hep G2肝癌细胞系(Hepatoblastoma G2 Cell Line,Hep G2 Cells)对Vγ9Vδ2T淋巴细胞免疫耐受中的作用及相关机制。方法:1.用CNOT7重组质粒(Recombinant plasmid of CNOT,sh CNOT7)及相应对照组载体质粒转染Hep G2细胞。2.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。3.Western blot方法检测各组细胞中CNOT7及信号传导及转录激活因子1(Signal Transducer and Activator of Transcription 1,STAT1)、STAT3的蛋白表达水平。4.Western blot法检测Hep G2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:1.使用Vγ9Vδ2T细胞因子干预后,CNOT7基因敲减后的Hep G2细胞组凋亡率由(7.55±2.63)%增加至(20.59±3.12)%。2.与L02细胞组比较,Hep G2细胞组中的CNOT7蛋白高表达(F=28.76,P0.01),STAT3蛋白高表达(F=110.29,P0.01),STAT1蛋白低表达(F=35.67,P0.01)。3.CNOT7基因敲除后,Hep G2细胞组的STAT1蛋白表达上调(t=6.69,P0.05)。结论:1.CNOT7基因参与诱导了Hep G2细胞对Vγ9Vδ2T细胞的免疫耐受,这与CNOT7和STAT3的过表达和STAT1的表达抑制相关联。2.敲减CNOT7基因可以通过上调STAT1蛋白的表达从而逆转Hep G2细胞对Vγ9Vδ2T细胞的免疫耐受。
[Abstract]:Aim: to investigate the role of human CCR4-NOT transcriptional complex subunit 7 (CCR4-NOT transcription complex subunit7 human,CNOT7) in the immune tolerance of Hep G2 Cell Line,Hep G2 Cells to V 纬 9V 未 2T lymphocytes. Methods: 1. Hep G2 cells were transfected with CNOT7 recombinant plasmid (Recombinant plasmid of CNOT,sh CNOT7 and corresponding control plasmid. 2. V 纬 9V 未 2T cytokines were used to interfere with each group of cells before and after transfection. Flow cytometry was used to detect apoptosis. 3.Western blot method was used to detect the expression of CNOT7, signal transduction and transcriptional activator 1 (STAT1), and STAT3 protein expression. 4.Western blot method was used to detect Hep G2 hepatoma cell line and human normal hepatocytes. The expression level of CNOT7,STAT1 and STAT3 protein in line L02. The result is 1: 1. After intervention with V 纬 9V 未 2T cytokines, the apoptosis rate of Hep G2 cells after CNOT7 gene knockout increased from (7.55 卤2.63)% to (20.59 卤3.12)%. Compared with L02 cell group, the expression of CNOT7 protein was higher in Hep G2 cell group than that in L02 cell group (Fnr 28.76 P0.01), STAT3 protein was highly expressed (Fnil 110.29, P0.01), and STAT1 protein was low (Ff35.67 P0.01). After 3.CNOT7 gene knockout, STAT1 protein expression in Hep G2 cell group was up-regulated (t6.69 P05). Conclusion: 1.CNOT7 gene is involved in inducing the immune tolerance of Hep G2 cells to V 纬 9V 未 2T cells, which is related to the overexpression of CNOT7 and STAT3 and the inhibition of STAT1 expression. 2. Knockout of CNOT7 gene can reverse the immune tolerance of Hep G2 cells to V 纬 9V 未 2T cells by up-regulating the expression of STAT1 protein.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7
本文编号：2286927
[Abstract]:Aim: to investigate the role of human CCR4-NOT transcriptional complex subunit 7 (CCR4-NOT transcription complex subunit7 human,CNOT7) in the immune tolerance of Hep G2 Cell Line,Hep G2 Cells to V 纬 9V 未 2T lymphocytes. Methods: 1. Hep G2 cells were transfected with CNOT7 recombinant plasmid (Recombinant plasmid of CNOT,sh CNOT7 and corresponding control plasmid. 2. V 纬 9V 未 2T cytokines were used to interfere with each group of cells before and after transfection. Flow cytometry was used to detect apoptosis. 3.Western blot method was used to detect the expression of CNOT7, signal transduction and transcriptional activator 1 (STAT1), and STAT3 protein expression. 4.Western blot method was used to detect Hep G2 hepatoma cell line and human normal hepatocytes. The expression level of CNOT7,STAT1 and STAT3 protein in line L02. The result is 1: 1. After intervention with V 纬 9V 未 2T cytokines, the apoptosis rate of Hep G2 cells after CNOT7 gene knockout increased from (7.55 卤2.63)% to (20.59 卤3.12)%. Compared with L02 cell group, the expression of CNOT7 protein was higher in Hep G2 cell group than that in L02 cell group (Fnr 28.76 P0.01), STAT3 protein was highly expressed (Fnil 110.29, P0.01), and STAT1 protein was low (Ff35.67 P0.01). After 3.CNOT7 gene knockout, STAT1 protein expression in Hep G2 cell group was up-regulated (t6.69 P05). Conclusion: 1.CNOT7 gene is involved in inducing the immune tolerance of Hep G2 cells to V 纬 9V 未 2T cells, which is related to the overexpression of CNOT7 and STAT3 and the inhibition of STAT1 expression. 2. Knockout of CNOT7 gene can reverse the immune tolerance of Hep G2 cells to V 纬 9V 未 2T cells by up-regulating the expression of STAT1 protein.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.7
【参考文献】
相关期刊论文 前3条
1 郭玲;何静;陈东风;;细胞免疫治疗原发性肝癌的现状及展望[J];实用肝脏病杂志;2015年06期
2 邱福铭;李忠朋;黄建;;γδT细胞为基础的抗肿瘤免疫治疗研究进展[J];浙江大学学报(医学版);2010年04期
3 赵海涛;李宁;张雪芳;毛一雷;桑新亭;黄洁夫;王kH;;肝癌免疫治疗研究进展[J];中华普通外科杂志;2008年07期
，本文编号：2286927
本文链接：https://www.wllwen.com/yixuelunwen/zlx/2286927.html
