DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species.
DNA double-strand breaks in the Toxoplasma gondii-infected cells by the action of reactive oxygen species.
复制标题
活性氧作用下弓形虫感染细胞中 DNA 双链断裂
DOI：
10.1186/s13071-020-04324-7
复制
发表时间：
2020-09-25
影响因子：
3.2
通讯作者：
Yang Y
中科院分区：
文献类型：
--
作者：
Zhuang H;Yao C;Zhao X;Chen X;Yang Y;Huang S;Pan L;Du A;Yang Y
刚地弓形虫是全球所有温血动物的专性寄生虫。一旦感染细胞，它就会操纵宿主的DNA损伤反应，这一点尚待阐明。本研究的目的有三个：(i)评估体外感染弓形虫细胞的DNA损伤；（ii）查明感染弓形虫的细胞中DNA损伤的原因；（iii）研究弓形虫感染期间DNA损伤反应的激活。HeLa、Vero和HEK293细胞均感染弓形虫，感染倍数（MOI）为10:1。在感染后10小时、20小时和30小时，用western blot和免疫荧光法分析感染细胞的DNA双链断裂(DSBs) γ - h2ax生物标志物。用2′，7′-二氯二氢荧光素（H2DCFDA）测定活性氧（ROS）水平，用活性氧抑制剂n -乙酰半胱氨酸（NAC）抑制ROS诱导的DNA损伤。最后，通过western blot检测共济失调毛细血管扩张突变/检查点激酶2 （ATM/CHK2）的活性形式来评估DNA损伤反应。与未感染的细胞相比，感染的HeLa细胞中的γ - h2ax水平在弓形虫感染期间随时间显著增加。NAC处理大大降低了宿主细胞中的ROS，同时降低了γ - h2ax。在弓形虫感染的细胞中，磷酸化的ATM/CHK2水平升高。弓形虫感染在体外引发宿主细胞中以ROS为主要参与者的DNA dsb。它还激活了DNA损伤反应通路ATM/CHK2。刚地弓形虫设法在其宿主细胞的生存和凋亡之间保持平衡，以有利于其自身的生存。
Toxoplasma gondii is an obligate parasite of all warm-blooded animals around the globe. Once infecting a cell, it manipulates the host’s DNA damage response that is yet to be elucidated. The objectives of the present study were three-fold: (i) to assess DNA damages in T. gondii-infected cells in vitro; (ii) to ascertain causes of DNA damage in T. gondii-infected cells; and (iii) to investigate activation of DNA damage responses during T. gondii infection. HeLa, Vero and HEK293 cells were infected with T. gondii at a multiplicity of infection (MOI) of 10:1. Infected cells were analyzed for a biomarker of DNA double-strand breaks (DSBs) γH2AX at 10 h, 20 h or 30 h post-infection using both western blot and immunofluorescence assay. Reactive oxygen species (ROS) levels were measured using 2′,7′-dichlorodihydrofluorescein diacetate (H2DCFDA), and ROS-induced DNA damage was inhibited by a ROS inhibitor N-acetylcysteine (NAC). Lastly, DNA damage responses were evaluated by detecting the active form of ataxia telangiectasia mutated/checkpoint kinase 2 (ATM/CHK2) by western blot. γH2AX levels in the infected HeLa cells were significantly increased over time during T. gondii infection compared to uninfected cells. NAC treatment greatly reduced ROS and concomitantly diminished γH2AX in host cells. The phosphorylated ATM/CHK2 were elevated in T. gondii-infected cells. Toxoplasma gondii infection triggered DNA DSBs with ROS as a major player in host cells in vitro. It also activated DNA damage response pathway ATM/CHK2. Toxoplasma gondii manages to keep a balance between survival and apoptosis of its host cells for the benefit of its own survival.
登录
查看更多内容
影响因子：
8
作者：
Pietsch, E. C.;Sykes, S. M.;McMahon, S. B.;Murphy, M. E.
通讯作者：
Murphy, M. E.
影响因子：
7.4
作者：
ARUOMA, OI;HALLIWELL, B;BUTLER, J
通讯作者：
BUTLER, J
影响因子：
4.6
作者：
Chniguir, Amina;Pintard, Coralie;Bachoual, Rafik
通讯作者：
Bachoual, Rafik
影响因子：
5.2
作者：
Chen Y;Williams V;Filippova M;Filippov V;Duerksen-Hughes P
通讯作者：
Duerksen-Hughes P
SFRS11 缺失通过调节 LRP8 和 ApoE 导致与衰老相关的认知能力下降
影响因子：
8.8
作者：
Raihan, Obayed;Brishti, Afrina;Liu, Qiang
通讯作者：
Liu, Qiang