TRAF1以及CD30-TRAF1信号通路在B细胞性霍奇金淋巴瘤细胞中的作用分析

Analysis on the significance of TRAF1 and CD30-TRAF1 signal pathway in B cells Hodgkin's lymphoma cells

目的探讨B细胞性霍奇金淋巴瘤细胞中肿瘤坏死因子受体相关因子1(TRAF1)和CD30-TRAF1信号通路的作用。方法选取B细胞性霍奇金淋巴瘤细胞株L428进行培养,采用荧光实时定量(RT-PCR)检测L428细胞TRAF1 mRNA表达情况,Western blot检测TRAF1蛋白表达水平,同时检测CD30配体对TRAF1的影响以及TRAF1对L428细胞凋亡的影响。结果 L428细胞TRAF1 mRNA和蛋白相对表达量分别为3.64±1.03和0.97±0.22,明显高于对照细胞的0.72±0.21和0.21±0.09,差异有统计学意义(P<0.05);通过荧光实时定量RT-PCR检测,L428细胞TRAF1 mRNA和蛋白相对表达量在CD30L作用前分别为3.62±0.34和2.29±0.27,CD30L作用8 h后增加为6.54±0.28和4.12±0.36,差异有统计学意义(P<0.05);转染TRAF1 siRNA后,L428细胞凋亡率为(29.17±6.38)%,明显高于转染对照的(4.27±1.06)%,差异有统计学意义(P<0.05);转染对照TRAF1蛋白相对表达为2.44±0.94,明显高于转染TRAF1 siRNA L428细胞的0.17±0.04,差异有统计学意义(P<0.05)。结论 B细胞性霍奇金淋巴瘤细胞中TRAF1表达增加,可能参与了霍奇金淋巴瘤细胞的抗凋亡过程,受到CD30信号通路的调控。

Objective To explore the role of tumor necrosis factor（ TNF） receptor associated factor 1（ TRAF1） and CD30- TRAF1 signaling pathway in B cell Hodgkin＇s lymphoma. Methods B- cell Hodgkin lymphoma cell line L428 had been selected and cultured,the expression of TRAF1 mRNA in L428 cells was detected by real- time fluorescence quantitative RT- PCR,and the expression level of TRAF1 protein was detected by Western blot,at the same time,the effect of CD30 ligand on TRAF1 and TRAF1 on apoptosis of L428 cells had been detected.Results TRAF1 mRNA in L428 cells and the quantity of protein expression were 3. 64 ± 1. 03 and 0. 97 ± 0. 22 respectively,they were significantly higher than those（ 0. 72 ± 0. 21 and 0. 21 ± 0. 09） in control cells,and the difference was statistically significant（ P〈0. 05）. By real-time fluorescence quantitative RT- PCR detection,TRAF1 mRNA in L428 cells and the relative expression quantity of protein before the action of CD30 L were 3. 62 ± 0. 34 and 2. 29 ± 0. 27 respectively,after the action of CD30 L for 8h,they were increased to 6. 54 ± 0. 28 and 4. 12 ± 0. 36 respectively,and the difference was statistically significant（ P〈0. 05）. TRAF1 siRNA after transfection,the rate of apoptosis in L428 cells was29. 17 ± 6. 38 %,it was significantly higher than that（ 4. 27 ± 1. 06%） of transfection controls,and the difference was statistically significant（P〈0. 05）. Transfection control TRAF1 protein expression was 2. 44 ± 0. 94,it was significantly higher than that（ 0. 17 ± 0. 04） of transfection with TRAF1 siRNA L428 cells（ P〈0. 05）. Conclusion The increased expression of TRAF1 in B cells Hodgkin＇s lymphoma cells may be involved in the anti- apoptotic process of Hodgkin＇s lymphoma cells,and it is regulated by CD30 signaling pathway.