JZUS - Journal of Zhejiang University SCIENCE

Journal of Zhejiang University SCIENCE B 2018 Vol.19 No.6 P.415-424

PGC-1α coordinates with Bcl-2 to control the cell cycle in U251 cells through reducing ROS

Author(s): Kun Yao, Xu-feng Fu, Xing Du, Yan Li, Shan-shan Yang, Min Yu, Qing-hua Cui
Affiliation(s): School of Life Sciences, Yunnan University, Kunming 650091, China; more
Corresponding email(s): cuiqinghua@ynu.edu.cn
Key Words: B-cell lymphoma 2 (Bcl-2), Peroxisome proliferator-activated receptor-γ, co-activator 1α, (PGC-1α, ), Mitochondria, Reactive oxygen species (ROS), Cell cycle

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Abstract: B-cell lymphoma 2 (Bcl-2) has a dual function, acting as both an oncogene and an anti-tumor gene. It is well known that Bcl-2 exerts its tumor promoting function through the mitochondrial pathway. However, the mechanism by which it suppresses tumor formation is not well understood. We have previously shown that Bcl-2 inhibits cell cycle progression from the G₀/G₁ to the S phase after serum starvation, and that quiescent Bcl-2 expressing cells maintain a significantly lower level of mitochondrial reactive oxygen species (ROS) than control cells. Based on the fact that ROS mediate cell cycle progression and are controlled by peroxisome proliferator-activated receptor-γ; co-activator 1α; (PGC-1α;), a key molecule induced by prolonged starvation and involved in mitochondrial metabolism, we hypothesized that PGC-1α might be related to the cell cycle function of Bcl-2. In this paper, we show that PGC-1α is upregulated by Bcl-2 overexpression and downregulated following Bcl-2 knockdown or downregulation after serum starvation. However, Bcl-2 is negatively regulated by PGC-1α expression. Further, co-immunoprecipitation (co-IP) experiments showed that PGC-1α protein is co-precipitated with Bcl-2 at the G₀/G₁ phase. Taken together, our results suggest that PGC-1α interacts with Bcl-2 after serum depletion, and that Bcl-2 might recruit PGC-1α to reduce ROS, which in turn delays cell cycle progression in coordination with Bcl-2.

PGC-1α在U251细胞中协同Bcl-2通过降低ROS来调控细胞周期

目的:探究过氧化物酶体增生激活受体γ协同刺激因子1α (PGC-1α)和B细胞淋巴瘤-2(Bcl-2)在调控细胞周期中的相互关系.
创新点:首次证明在血清饥饿时PGC-1α负调控Bcl-2,并且认为Bcl-2可能通过招募PGC-1α降低细胞中的活性氧自由基(ROS)以调节细胞周期.
方法:用蛋白质印迹法(Western blotting)检测了接触抑制和血清饥饿处理的NIH3T3过表达Bcl-2的细胞中PGC-1α的表达,并且分别检测了用Bcl-2和PGC-1α的小干扰RNA(siRNA)降低U251细胞(内源性高表达Bcl-2和PGC-1α)中的Bcl-2和PGC-1α的表达,最后用免疫共沉淀(co-IP)检测了二者的关系.
结论:本实验中用两种细胞同步化的方法(接触抑制和血清饥饿)处理了Bcl-2过表达的NIH3T3细胞时发现PGC-1α高表达,用Bcl-2的siRNA处理了U251细胞时发现PGC-1α的表达降低,但是血清饥饿处理了U251后发现Bcl-2升高而PGC-1α降低,而且PGC-1α被siRNA降低后Bcl-2反而上升,最后用Bcl-2抗体免疫共沉淀了PGC-1α蛋白,这些结果说明在血清饥饿时PGC-1α负调控Bcl-2行使调节细胞周期的功能.
关键词：过氧化物酶体增生激活受体γ协同刺激因子1α (PGC-1α);B细胞淋巴瘤-2(Bcl-2);线粒体;活性氧自由基(ROS);细胞周期

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[28]List of electronic supplementary materials

[29]Fig. S1 Immunofluorescence images of PGC-1α and Bcl-2 protein co-localization in NIH3T3 cells

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PGC-1α在U251细胞中协同Bcl-2通过降低ROS来调控细胞周期

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