基于iTRAQ技术的伴放线聚集杆菌细胞致死性扩张毒素诱导Jurkat细胞的凋亡研究

Study of Aggregatibacter actinomycetemcomitans cytolethal distending toxin on inducing apoptosis in human Jurkat cells by iTRAQ labeling technique

目的探索伴放线聚集杆菌(Aggregatibacter actinomycetemcomitans,A.a)的一种毒力因子(cytolethal distending toxin,CDT)诱导人T淋巴细胞发生凋亡过程中表达改变的信号分子。方法培养Jurkat细胞,并进行分组:不做处理的正常细胞对照组、野生型A.a CDT刺激Jurkat细胞的野生组、突变型A.a CDT刺激Jurkat细胞的突变组,应用i TRAQ技术检测A.a CDT作用Jurkat细胞24 h过程中表达差异蛋白。结果通过筛选,我们得到20个凋亡相关的蛋白(变化倍数>1.3),其中表达上调的有8个(CASP8/CD3E/YY1/SH3BGRL3/P53/CASP3/UBE2I/TNFRSF10B),表达下调的有12个(F5/RPL18A/DAD1/MTCH2/HIST1H1E/CNBP/RBM25/SLC16A1/KDM2A/CD99/RAC2/TMX1)。结论本研究检测出了伴放线聚集杆菌细胞致死性膨胀毒素诱导人T淋巴细胞凋亡过程中表达差异的蛋白,并筛选了一条可能的信号通路UBE2I/P53/TNFRSF10B/CASP8/CASP3,为下一步研究及临床治疗局限型侵袭性牙周炎提供思路。

Objective To explore the key signaling molecules when cytolethal distending toxin （CDT）, a kind of Aggregatibacter ac- tinomyceterncomitans （A.a） induced apoptosis of T lymphoeytes in vitro. Methods Jurkat cells were cultured and then divided into groups： the control group, the wild group （Jurkat cells were stimulated by wild A. a CDT）, and the mutant group （Jurkat cells were stimulated by mutant A.a CDT）. After human Jurkat cells were treated by A. a CDT for 24 hours, iTRAQ （ Isobaric tags for relative and absolute quantitation） labeling technique was used to test differentially expressed apoptosis-relating proteins. Results 20 apoptosis-re- lated differentially expressed proteins （fold change ratio〉 1.3 ） were identified, including 8 proteins （CASP8/CD3E/YY1/SH3BGRL3/ P53/CASP3/UBE2I/ TNFRSF10B） which were upregnlated and 12 proteins （FS/RPL18A/DAD1/MTCH2/HIST1H1E/CNBP/ RBM25/SLC16A1/KDM2A/CD99/RAC2/TMX1） which were downregulated. Conclusion This study elucidated the signaling mole- cules when A.a CDT induced apoptosis of human Jurkat cells at the molecular level, and one possible signaling pathway was selected （UBE2I/P53/TNFRSFlOB/CASP8/CASP3）, providing thoughts for the future studies and the treatment of localized aggressive peri- odontitis.