BCL11B在细菌外毒素SEA刺激T细胞NF-κB分子表达中的作用研究

发布时间：2018-03-05 22:27

本文选题：SEA　切入点：BCL11B　出处：《暨南大学》2015年硕士论文　论文类型：学位论文

【摘要】：目的:研究金黄色葡萄球菌肠毒素A(SEA)对T细胞TCR信号通路中BCL11B、NF-κB分子转录与蛋白表达的影响;探讨细菌外毒素超抗原和T细胞介导的炎症、T细胞白血病发生之间的可能关系,为阐述淋巴细胞白血病发病机制提供实验数据。方法:1.实时荧光定量PCR(q RT-PCR)检测BCL11B以及NF-κB基因表达。2.免疫印迹(Western blotting)检测BCL11B以及NF-κB蛋白表达。3.BCL11B-si RNA转染技术沉默BCL11B,结合q RT-PCR与Western blotting技术。结果:1.在1μg/m LSEA作用下,Jurkat细胞中的BCL11B以及NF-κB基因m RNA的表达量随着刺激时间(0、15、45、180、360 min)的增加明显上调,且BCL11B基因m RNA的表达量明显大于NF-κBm RNA表达量(P0.05)。2.在20ng/m L浓度乙酸肉豆蔻佛波醇(PMA)联合1μM离子霉素(IO)作用下,Jurkat细胞中的BCL11B以及NF-κB基因m RNA的表达量随着刺激时间(0、15、45、180、360 min)的增加明显增加(P0.05),两者变化类似SEA刺激结果。3.在20ng/m L TNF-α作用下,Jurkat细胞中的NF-κB基因m RNA的表达量随着时间(0、15、45、180、360 min)的增加明显上调(P0.05);但是Jurkat细胞中的BCL11B基因m RNA的表达量没有明显变化。4.在1μg/m L SEA作用下,BCL11B、NF-ΚB分子的蛋白表达量随刺激时间(0、15、45、180、360 min)而明显增加(P0.05),与m RNA水平类似。5.在20ng/m L PMA联合1μM IO作用下,BCL11B、NF-ΚB分子的蛋白表达量和刺激时间有关系(P0.05),随时间(0、15、45、180、360 min)的增加逐步增加,与m RNA水平类似。6.在20ng/m L TNF-α作用下,BCL11B基因蛋白表达量无明显变化;NF-ΚB基因蛋白的表达量随刺激时间(0、15、45、180、360 min)的增加明显增加(P0.05),与m RNA水平类似。7.BCL11B-si RNA转染Jurkat细胞48小时后,以1μg/m L的SEA刺激Jurkat细胞(0、15、45、180、360 min)后,NF-κB基因蛋白表达无明显改变(P0.05)。8.BCL11B-si RNA转染Jurkat细胞48小时后,以20ng/m L的PMA和1u M的IO刺激Jurkat细胞0、15、45、180、360 min后,NF-κB基因蛋白表达量无明显改变(P0.05)。结论:1.细菌外毒素SEA可以刺激T细胞上调BCL11B基因、蛋白水平的表达,以及NF-κB基因、蛋白水平的表达。2.SEA以超抗原的方式,通过TCR途径刺激BCL11B基因、蛋白水平的表达、以及NF-κB基因、蛋白水平的表达。3.超抗原SEA可以通过TCR途径诱导BCL11B基因的表达而影响NF-κB的表达水平。
[Abstract]:Objective: to study the effects of staphylococcal enterotoxin Agna on the transcription and protein expression of BCL11BnNF- 魏 B in T cell TCR signaling pathway, and to explore the possible relationship between bacterial exotoxin superantigen and T cell-mediated inflammatory T cell leukemia. To provide experimental data to elucidate the pathogenesis of lymphocytic leukemia. Methods: 1. Real-time fluorescence quantitative PCR(q RT-PCR was used to detect BCL11B and NF- 魏 B gene expression .2. Western blotting was used to detect BCL11B and NF- 魏 B protein expression. 3. BCL11B-si RNA transfection technique was used to silence BCL11B. Q RT-PCR and Western blotting techniques. Results: 1. The expression of BCL11B and NF- 魏 B gene m RNA in Jurkat cells increased significantly with the stimulation time of 1 渭 g / m LSEA. The expression of BCL11B gene m RNA was significantly higher than that of NF- 魏 B m RNA. The expression of BCL11B and NF- 魏 B gene m RNA in Jurkat cells was significantly higher than that of NF- 魏 B m RNA. The expression of BCL11B and NF- 魏 B gene m RNA in Jurkat cells was significantly higher than that of NF- 魏 B RNA at the concentration of 20ng-1 / ml acetate combined with 1 渭 M ionomycin. The expression of BCL11B and NF- 魏 B gene m RNA in Jurkat cells increased with the time of stimulation. The expression of NF- 魏 B gene m RNA in Jurkat cells increased significantly with the increase of time, but the expression of BCL11B gene m RNA in Jurkat cells was significantly increased with the increase of P0.05min. However, the expression of BCL11B gene m RNA in Jurkat cells was similar to that of Jurkat cells induced by TNF- 伪. The expression of BCL11B gene m RNA in Jurkat cells was significantly increased with the increase of P0. 05%, but the expression of BCL11B gene m RNA in Jurkat cells was significantly increased with the increase of P0. 05%; however, the expression of BCL11B gene m RNA in Jurkat cells was significantly increased with time. The protein expression of BCL11BNF-KB molecule increased significantly with the stimulation time of 1 渭 g / mL SEA, which was similar to that of m RNA. The protein expression and stimulation time of BCL11BNF-KB molecule were similar to that of m RNA. The protein expression and stimulation time of BCL11BNF-KB molecule were observed in the presence of 20ng / mL PMA and 1 渭 MIO. The increase of P0. 05% is gradually increased with the increase of 0 / 15 / 45 / 180 / 360 / min. The protein expression of BCL11B gene was similar to that of m RNA. There was no significant change in the protein expression of BCL11B gene under the action of 20ng / mL TNF- 伪. The increase of protein expression of NF-KB gene increased significantly with the stimulation time. The expression of BCL11B gene protein increased significantly after transfection of Jurkat cells with Jurkat cells 48 hours after transfection with Jurkat cells, which was similar to the level of m RNA. 7. BCL11B-si RNA was transfected into Jurkat cells for 48 hours. After 1 渭 g / mL SEA was used to stimulate Jurkat cells, the expression of NF- 魏 B gene protein did not change significantly after transfection of Jurkat cells with BCL11B-si RNA for 48 hours. The expression of NF- 魏 B protein in Jurkat cells stimulated with 20ng / mL PMA and 1u M IO was not significantly changed after the stimulation of Jurkat cells with 20ng / mL PMA and 1u M IO. Conclusion: 1. Bacterial exotoxin SEA can up-regulate the expression of BCL11B gene, protein level and NF- 魏 B gene in T cells, and increase the expression of BCL11B gene, protein level and NF- 魏 B gene in T cells. Protein level expression. 2. Sea stimulates the expression of BCL11B gene, protein level and NF- 魏 B gene through TCR pathway in superantigen manner. Protein expression. 3. Superantigen SEA can induce the expression of BCL11B gene through TCR pathway and affect the expression level of NF- 魏 B.
【学位授予单位】：暨南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R733.7

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