Research On The Role Of BCL11B In NF-κ B Expression After Bacterial Exotoxin SEA Stimulates T Cells

Objective:To investigate the effects of bacterial exotoxinstaphylococcal enterotoxins A(SEA) on BCL11 B or NF-κB genes transcriptions and proteins expressions in TCR signaling pathway in the T cells; To explore the potential relationship of the bacterial exotoxin superantigen and inflammation or human T-cell leukemia, to provide the experimental datas for exploring the pathogenesis of lymphocytic leukemia.Methods:1. To detect the expression of BCL11 B or NF-κB gene by q RT-PCR.2. To detect the expression of BCL11 B or NF-κB protein by Western blotting.3. BCL11 B genes were silenced by BCL11B-si RNA, which combined with q RT-PCR and Western blotting.Results:1. The expression of BCL11 B or NF-κB m RNA were up- regulated over time(0、15、45、180、360 min), and the expression of BCL11 B m RNA was obviously stronger than NF-κB(P<0.05),when Jurkat cells were stimulated with 1μg/m L SEA.2. The expression of BCL11 B or NF-κB m RNA were up- regulated over time(0、15、45、180、360 min)(P<0.05), when Jurkat cells were stimulated with 20ng/m L PMA and 1μM IO, the results were similar with the consequence of Jurkat cells stimulated by SEA.3. The expression of NF-κB m RNA was up- regulated over time(0、15、45、180、360 min)(P<0.05), but the expression of BCL11 B m RNA was not obvious,when Jurkat cells were stimulated with 20ng/m L TNF-α.4. The expression of BCL11 B or NF-κB protein were up- regulated over time(0、15、45、180、360 min)(P<0.05), when Jurkat cells were stimulated with 1μg/m L SEA, and the results were similar with the m RNA level.5. The expression of BCL11 B or NF-κB proteins were up-regulated over time(0、15、45、180、360 min)(P<0.05), when Jurkat cells were stimulated with 20ng/m L PMA and 1μM IO, the results were similar with the consequence of Jurkat stimulated by SEA.6. The expression of NF-κB protein was up-regulated over time(0、15、45、180、360 min)(P<0.05), but the expression of BCL11 B protein was not obvious,when Jurkat cells were stimulated with 20ng/m L TNF-α and the results were similar with the m RNA level.7. The expression of NF-κB protein was not obvious(P>0.05), when Jurkat cells were stimulated with 1μg/m L SEA(0、15、45、180、360 min)after transfected by BCL11B-si RNA 48 hours.8. The expression of NF-κB protein was not obvious(P>0.05), when Jurkat cells were stimulated with 20ng/m L PMA and 1μM IO(0、15、45、180、360 min)after transfected by BCL11B-si RNA 48 hours.Conclusions:1. The expression of NF-κB m RNA or protein, BCL11 B m RNA or protein were upregulated, when Jurkat cells stimulated by Bacterial exotoxin SEA.2. The expression of NF-κB m RNA or protein, BCL11 B m RNA or protein were stimulated by superantigen SEA through TCR pathway.3. Superantigen SEA induced the expression of BCL11 B gene than effected NF-κB expression.