呼吸道合胞病毒通过TRAF6/NF-κB通路调控人T淋巴细胞白血病细胞增殖  

Regulation of Jurkat cell proliferation by respiratory syncytial virus through TRAF6/NF-κB pathway

作　　者：聂钰君 侯燕 钱丹 NIE Yujun;HOU Yan;QIAN Dan(Department of Pediatrics,Xiangyang Central Hospital,Affiliated Hospital of Hubei University of Arts and Science,Xiangyang 441021,China)

机构地区：[1]湖北文理学院附属医院襄阳市中心医院,湖北襄阳441021

出　　处：《实用医学杂志》2022年第4期434-438,共5页The Journal of Practical Medicine

基　　金：湖北省儿科联盟科学基金资助项目(编号:HBPASF-2019-7)。

摘　　要：目的通过呼吸道合胞病毒(RSV)感染人Jurkat细胞(T淋巴细胞白血病细胞),观察Jurkat细胞增殖及凋亡情况,探讨RSV是否对Jurkat细胞具有溶瘤作用及可能机制。方法将RSV感染Jurkat细胞,设未感染RSV的Jurkat细胞为空白对照组(con),在感染后不同时间点(3、6、12、24、48 h)收集细胞。采用qRT-PCR检测RSV-NS2(呼吸道合胞病毒非结构蛋白2)以了解Jurkat细胞感染RSV情况,MTT、克隆形成实验检测Jurkat细胞的增殖,FCM检测Jurkat细胞的凋亡水平,qRT-PCR检测TRAF6(肿瘤坏死因子受体相关因子6)的表达,Western blot检测Bcl2、TRAF6、NF-κB和cleaved Caspase-3、8、9的表达。结果RSV感染Jurkat细胞,q RT-PCR检测到Jurkat细胞在感染后3 h内RSV-NS2表达开始出现,6 h明显增高,随时间延长而升高,差异有统计学意义(P<0.05),而TRAF6的表达随感染时间的延长而减少(P<0.05)。MTT结果显示RSV感染后6~24 h,Jurkat细胞的增殖能力显著下降(P<0.05),克隆形成实验示RSV感染Jurkat后12 d,Jurkat细胞的克隆数明显减少(P<0.05),提示RSV抑制了Jurkat细胞增殖。FCM结果显示RSV感染后12~48 h后,Jurkat细胞的凋亡水平显著上升,呈时间依赖性(P<0.05)。Western blot的结果显示,RSV感染12 h以后,cleaved Caspase-3和cleaved Caspase-9的表达均显著增加,Bcl2的表达明显减少,有时间依赖性(P<0.05),cleaved Caspase-8的表达无明显改变,TRAF6和p-NF-κB的表达随感染时间的延长而降低(P<0.05)。结论降低RSV抑制Jurkat细胞增殖,促进细胞凋亡,通过抑制TRAF6/NF-κB通路发挥对Jurkat细胞的抑制作用,揭示RSV对血液系统肿瘤能产生溶瘤效应。Objective To observe the proliferation and apoptosis of Jurkat cells,and to discuss the oncolytic effect of RSV on Jurkat cells and its possible mechanism.Refractory or recurrent acute lymphoblastic leukemia is an important role affecting the overall efficacy of children with leukemia.Oncolytic virotherapy is a new biological therapy against tumors in recent years.Multiple oncolytic viruses,such as measel virus and Newcastle virus,have been widely researched in the treatment of leukemia.However,there are few studies in the treatment of leukemia with respiratory syncytial virus(RSV).Methods Jurkat cells were infected with RSV.Jurkat cells not infected with RSV were set as the blank control group(con).Cells were collected at different time points(3 h,6 h,12 h,24 h and 48 h)after infection.Respiratory syncytial virus nonstructural protein 2(RSV-NS2)and TRAF6 were detected by qRT-PCR.The proliferation of Jurkat cells was evaluated by MTT and clone formation assay,and the apoptosis level was evaluated by FCM.The protein expression of Bcl2 and TRAF6 and NF-κB and cleaved Caspase-3,8,9 was compared by Western blot.Results After RSV infected Jurkat cells,qRT-PCR detected that the expression of RSV-NS2 began to appear within 3 h after infection,increased significantly at 6 h,then increased in a timedependent manner(P<0.05),while the expression of TRAF6 decreased with the extension of infection time(P<0.05).MTT results showed that the proliferation of Jurkat cells decreased significantly from 6 h to 24 h after RSV infection(P<0.05).Clone formation experiment showed that the number of clones of Jurkat cells decreased significantly 12 days after RSV infection(P<0.05),suggesting that RSV inhibited the proliferation of Jurkat cells.FCM results showed that the apoptosis level of Jurkat cells increased significantly in a time-dependent manner from 12 h to 48 h after RSV infection(P<0.05).Western blot analysis revealed that the expression of cleaved caspase-3 and cleaved caspase-9 increased significantly after 12 hours of RSV in

关 键 词：呼吸道合胞病毒 JURKAT TRAF6 NF-ΚB 

分 类 号：R733.7[医药卫生—肿瘤]