【摘要】：目的:研究人CCR4-NOT轉(zhuǎn)錄復(fù)合體亞基7(CCR4-NOT transcription complex subunit7 human,CNOT7)在Hep G2肝癌細(xì)胞系(Hepatoblastoma G2 Cell Line,Hep G2 Cells)對Vγ9Vδ2T淋巴細(xì)胞免疫耐受中的作用及相關(guān)機(jī)制。方法:1.用CNOT7重組質(zhì)粒(Recombinant plasmid of CNOT,sh CNOT7)及相應(yīng)對照組載體質(zhì)粒轉(zhuǎn)染Hep G2細(xì)胞。2.用Vγ9Vδ2T細(xì)胞因子干預(yù)轉(zhuǎn)染前后的各組細(xì)胞,流式細(xì)胞儀檢測細(xì)胞凋亡水平。3.Western blot方法檢測各組細(xì)胞中CNOT7及信號傳導(dǎo)及轉(zhuǎn)錄激活因子1(Signal Transducer and Activator of Transcription 1,STAT1)、STAT3的蛋白表達(dá)水平。4.Western blot法檢測Hep G2肝癌細(xì)胞系及人正常肝細(xì)胞系L02中CNOT7、STAT1和STAT3蛋白的表達(dá)水平。結(jié)果:1.使用Vγ9Vδ2T細(xì)胞因子干預(yù)后,CNOT7基因敲減后的Hep G2細(xì)胞組凋亡率由(7.55±2.63)%增加至(20.59±3.12)%。2.與L02細(xì)胞組比較,Hep G2細(xì)胞組中的CNOT7蛋白高表達(dá)(F=28.76,P0.01),STAT3蛋白高表達(dá)(F=110.29,P0.01),STAT1蛋白低表達(dá)(F=35.67,P0.01)。3.CNOT7基因敲除后,Hep G2細(xì)胞組的STAT1蛋白表達(dá)上調(diào)(t=6.69,P0.05)。結(jié)論:1.CNOT7基因參與誘導(dǎo)了Hep G2細(xì)胞對Vγ9Vδ2T細(xì)胞的免疫耐受,這與CNOT7和STAT3的過表達(dá)和STAT1的表達(dá)抑制相關(guān)聯(lián)。2.敲減CNOT7基因可以通過上調(diào)STAT1蛋白的表達(dá)從而逆轉(zhuǎn)Hep G2細(xì)胞對Vγ9Vδ2T細(xì)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 緯 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 緯 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 緯 9V 未 2T cytokines, the apoptosis rate of Hep G2 cells after CNOT7 gene knockout increased from (7.55 鹵2.63)% to (20.59 鹵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 緯 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 緯 9V 未 2T cells by up-regulating the expression of STAT1 protein.
【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R735.7

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