發(fā)布時間：2018-03-05 22:27

  本文選題：SEA　切入點：BCL11B　出處：《暨南大學》2015年碩士論文　論文類型：學位論文

【摘要】：目的:研究金黃色葡萄球菌腸毒素A(SEA)對T細胞TCR信號通路中BCL11B、NF-κB分子轉錄與蛋白表達的影響;探討細菌外毒素超抗原和T細胞介導的炎癥、T細胞白血病發(fā)生之間的可能關系,為闡述淋巴細胞白血病發(fā)病機制提供實驗數據。方法:1.實時熒光定量PCR(q RT-PCR)檢測BCL11B以及NF-κB基因表達。2.免疫印跡(Western blotting)檢測BCL11B以及NF-κB蛋白表達。3.BCL11B-si RNA轉染技術沉默BCL11B,結合q RT-PCR與Western blotting技術。結果:1.在1μg/m LSEA作用下,Jurkat細胞中的BCL11B以及NF-κB基因m RNA的表達量隨著刺激時間(0、15、45、180、360 min)的增加明顯上調,且BCL11B基因m RNA的表達量明顯大于NF-κBm RNA表達量(P0.05)。2.在20ng/m L濃度乙酸肉豆蔻佛波醇(PMA)聯合1μM離子霉素(IO)作用下,Jurkat細胞中的BCL11B以及NF-κB基因m RNA的表達量隨著刺激時間(0、15、45、180、360 min)的增加明顯增加(P0.05),兩者變化類似SEA刺激結果。3.在20ng/m L TNF-α作用下,Jurkat細胞中的NF-κB基因m RNA的表達量隨著時間(0、15、45、180、360 min)的增加明顯上調(P0.05);但是Jurkat細胞中的BCL11B基因m RNA的表達量沒有明顯變化。4.在1μg/m L SEA作用下,BCL11B、NF-ΚB分子的蛋白表達量隨刺激時間(0、15、45、180、360 min)而明顯增加(P0.05),與m RNA水平類似。5.在20ng/m L PMA聯合1μM IO作用下,BCL11B、NF-ΚB分子的蛋白表達量和刺激時間有關系(P0.05),隨時間(0、15、45、180、360 min)的增加逐步增加,與m RNA水平類似。6.在20ng/m L TNF-α作用下,BCL11B基因蛋白表達量無明顯變化;NF-ΚB基因蛋白的表達量隨刺激時間(0、15、45、180、360 min)的增加明顯增加(P0.05),與m RNA水平類似。7.BCL11B-si RNA轉染Jurkat細胞48小時后,以1μg/m L的SEA刺激Jurkat細胞(0、15、45、180、360 min)后,NF-κB基因蛋白表達無明顯改變(P0.05)。8.BCL11B-si RNA轉染Jurkat細胞48小時后,以20ng/m L的PMA和1u M的IO刺激Jurkat細胞0、15、45、180、360 min后,NF-κB基因蛋白表達量無明顯改變(P0.05)。結論:1.細菌外毒素SEA可以刺激T細胞上調BCL11B基因、蛋白水平的表達,以及NF-κB基因、蛋白水平的表達。2.SEA以超抗原的方式,通過TCR途徑刺激BCL11B基因、蛋白水平的表達、以及NF-κB基因、蛋白水平的表達。3.超抗原SEA可以通過TCR途徑誘導BCL11B基因的表達而影響NF-κB的表達水平。
[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- 偽. 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- 偽. 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.
【學位授予單位】：暨南大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R733.7

【參考文獻】

相關期刊論文 前1條

1 黃欣;李揚秋;;BCL 11基因與淋巴細胞腫瘤[J];癌癥進展;2009年05期

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本文編號：1572190