Full Text:   <3277>

Summary:  <1612>

CLC number: 

On-line Access: 2024-08-27

Received: 2023-10-17

Revision Accepted: 2024-05-08

Crosschecked: 0000-00-00

Cited: 0

Clicked: 4370

Citations:  Bibtex RefMan EndNote GB/T7714

 ORCID:

Xing DU

https://orcid.org/0000-0003-2683-4524

Xiuying PEI

https://orcid.org/0000-0002-4410-8588

-   Go to

Article info.
Open peer comments

Journal of Zhejiang University SCIENCE B 2021 Vol.22 No.10 P.839-855

http://doi.org/10.1631/jzus.B2000802


A proteomic analysis of Bcl-2 regulation of cell cycle arrest: insight into the mechanisms


Author(s):  Xing DU, Jingjing XIAO, Xufeng FU, Bo XU, Hang HAN, Yin WANG, Xiuying PEI

Affiliation(s):  Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Medical University, Yinchuan 750004, China; more

Corresponding email(s):   yin-wang@hotmail.com, peixiuying@163.com

Key Words:  Cell cycle, B cell lymphoma 2 (Bcl-2), Proteomics, Oxidative phosphorylation


Xing DU, Jingjing XIAO, Xufeng FU, Bo XU, Hang HAN, Yin WANG, Xiuying PEI. A proteomic analysis of Bcl-2 regulation of cell cycle arrest: insight into the mechanisms[J]. Journal of Zhejiang University Science B, 2021, 22(10): 839-855.

@article{title="A proteomic analysis of Bcl-2 regulation of cell cycle arrest: insight into the mechanisms",
author="Xing DU, Jingjing XIAO, Xufeng FU, Bo XU, Hang HAN, Yin WANG, Xiuying PEI",
journal="Journal of Zhejiang University Science B",
volume="22",
number="10",
pages="839-855",
year="2021",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.B2000802"
}

%0 Journal Article
%T A proteomic analysis of Bcl-2 regulation of cell cycle arrest: insight into the mechanisms
%A Xing DU
%A Jingjing XIAO
%A Xufeng FU
%A Bo XU
%A Hang HAN
%A Yin WANG
%A Xiuying PEI
%J Journal of Zhejiang University SCIENCE B
%V 22
%N 10
%P 839-855
%@ 1673-1581
%D 2021
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2000802

TY - JOUR
T1 - A proteomic analysis of Bcl-2 regulation of cell cycle arrest: insight into the mechanisms
A1 - Xing DU
A1 - Jingjing XIAO
A1 - Xufeng FU
A1 - Bo XU
A1 - Hang HAN
A1 - Yin WANG
A1 - Xiuying PEI
J0 - Journal of Zhejiang University Science B
VL - 22
IS - 10
SP - 839
EP - 855
%@ 1673-1581
Y1 - 2021
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2000802


Abstract: 
B cell lymphoma 2 (Bcl-2) is an important antiapoptotic gene that plays a dual role in the maintenance of the dynamic balance between the survival and death of cancer cells. In our previous study, Bcl-2 was shown to delay the G0/G1 to S phase entry by regulating the mitochondrial metabolic pathways to produce lower levels of adenosine triphosphate (ATP) and reactive oxygen species (ROS). However, the detailed molecular mechanisms or pathways by which Bcl-2 regulates the cell cycle remain unknown. Here, we compared the effects of Bcl-2 overexpression with an empty vector control in the NIH3T3 cell line synchronized by serum starvation, and evaluated the effects using proteomic analysis. The effect of Bcl-2 on cell cycle regulation was detected by monitoring Bcl-2 and p27 expression. The result of subsequent proteomic analysis of Bcl-2 overexpressing cells identified 169 upregulated and 120 downregulated proteins with a 1.5-fold change. These differentially expressed proteins were enriched in a number of signaling pathways predominantly involving the ribosome and oxidative phosphorylation, according to the data of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. These results indicated that Bcl-2 potentially acts at the translation level to influence proteins or enzymes of the respiratory chain or in the ribosome, and thereby regulates the cell cycle. Additionally, differentially expressed proteins involved in oxidative phosphorylation were determined to account for most of the effects of Bcl-2 on the cell cycle mediated by the mitochondrial pathway investigated in our previous study. These results can provide assistance for additional in-depth studies on the regulation of the cell cycle by Bcl-2. The results of the proteomic analysis determined the mechanism of Bcl-2-dependent delay of the cell cycle progression. In summary, the results of this study provide a novel mechanistic basis for identifying the key proteins or pathways for designing and developing precisely targeted cancer drugs.

Bcl-2调控细胞周期阻滞的蛋白质组学分析及其机制研究

目的:B细胞淋巴瘤-2(Bcl-2)基因除了广为人知的抗凋亡功能之外,还具有调控细胞周期的非凋亡功能,但是机制却不清楚。作者前期研究发现Bcl-2可以通过阻滞G0/G1期进入S期的进程调控细胞周期,可能与低水平的三磷酸腺苷(ATP)和活性氧自由基(ROS)有关。因此,本研究旨在探究其潜在调控机制。
创新点:基于Bcl-2通过ATP和ROS调控细胞周期的前期发现,本研究首次利用蛋白组学方法系统研究了Bcl-2调控细胞周期的潜在机制。
方法:联合利用蛋白质印迹(western blotting)和蛋白质组学方法研究血清饥饿同步化处理的Bcl-2过表达和对照组细胞株,并结合蛋白组学中差异蛋白的基因本体(Gene Ontology,GO)和Kyoto Encyclopedia of Genes and Genomes(KEGG)分析,进一步明确Bcl-2调控细胞周期的潜在机制。
结论:蛋白组学结果显示,在1.5倍差异下共有169个蛋白发生了上调,120个蛋白发生了下调。通过GO和KEGG分析,这些差异蛋白富集到多个通路,主要集中在呼吸链和核糖体相关信号通路。这些结果表明Bcl-2可能在翻译水平影响核糖体和氧化磷酸化进而调控细胞周期。本研究为进一步靶向Bcl-2调控细胞周期抗癌药物研究了提供重要的理论基础。

关键词:细胞周期;B细胞淋巴瘤-2(Bcl-2);蛋白质组学;氧化磷酸化

Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article

Reference

[1]AdamsCM, Clark-GarveyS, PorcuP, et al., 2019. Targeting the Bcl-2 family in B cell lymphoma. Front Oncol, 8:636.

[2]Bonnefoy-BerardN, AouacheriaA, VerscheldeC, et al., 2004. Control of proliferation by Bcl-2 family members. Biochim Biophys Acta-Mol Cell Res, 1644(2-3):159-168.

[3]BradyHJ, Gil-GómezG, KirbergJ, et al., 1996. Baxα perturbs T cell development and affects cell cycle entry of T cells. EMBO J, 15(24):6991-7001.

[4]ClearyML, SmithSD, SklarJ, 1986. Cloning and structural analysis of cDNAs for bcl-2 and a hybrid bcl-2/immunoglobulin transcript resulting from the t(14;18) translocation. Cell, 47(1):19-28.

[5]DelbridgeARD, GrabowS, StrasserA, et al., 2016. Thirty years of BCL-2: translating cell death discoveries into novel cancer therapies. Nat Rev Cancer, 16(2):99-109.

[6]del Gaizo MooreV, LetaiA, 2013. BH3 profiling—measuring integrated function of the mitochondrial apoptotic pathway to predict cell fate decisions. Cancer Lett, 332(2):202-205.

[7]DuX, FuXF, YaoK, et al., 2017. Bcl-2 delays cell cycle through mitochondrial ATP and ROS. Cell Cycle, 16(7):707-713.

[8]FrantziM, LatosinskaA, MischakH, 2019. Proteomics in drug development: the dawn of a New Era? Proteomics Clin Appl, 13(2):e1800087.

[9]FuXF, YaoK, DuX, et al., 2016. PGC-1α regulates the cell cycle through ATP and ROS in CH1 cells. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 17(2):136-146.

[10]GaoSW, LiuF, 2019. Novel insights into cell cycle regulation of cell fate determination. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(6):467-475.

[11]GrossA, KatzSG, 2017. Non-apoptotic functions of BCL-2 family proteins. Cell Death Differ, 24(8):1348-1358.

[12]HatokJ, RacayP, 2016. Bcl-2 family proteins: master regulators of cell survival. Biomol Concepts, 7(4):259-270.

[13]HuSY, ZhuangQQ, QiuY, et al., 2019. Cell models and drug discovery for mitochondrial diseases. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 20(5):449-456.

[14]JanumyanYM, SansamCG, ChattopadhyayA, et al., 2003. Bcl-xL/Bcl-2 coordinately regulates apoptosis, cell cycle arrest and cell cycle entry. EMBO J, 22(20):5459-5470.

[15]JanumyanYM, CuiQH, YanL, et al., 2008. G0 function of BCL2 and BCL-xL requires BAX, BAK, and p27 phosphorylation by Mirk, revealing a novel role of BAX and BAK in quiescence regulation. J Biol Chem, 283(49):34108-34120.

[16]JardimFR, de AlmeidaFJS, LuckachakiMD, et al., 2020. Effects of sulforaphane on brain mitochondria: mechanistic view and future directions. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 21(4):263-279.

[17]KaleJ, OsterlundEJ, AndrewsDW, 2018. BCL-2 family proteins: changing partners in the dance towards death. Cell Death Differ, 25(1):65-80.

[18]KampenKR, SulimaSO, VerbelenB, et al., 2019. The ribosomal RPL10 R98S mutation drives IRES-dependent BCL-2 translation in T-ALL. Leukemia, 33(2):319-332.

[19]KimHJ, MaitiP, BarrientosA, 2017. Mitochondrial ribosomes in cancer. Semin Cancer Biol, 47:67-81.

[20]KroemerG, GalluzziL, BrennerC, 2007. Mitochondrial membrane permeabilization in cell death. Physiol Rev, 87(1):99-163.

[21]LessardF, Brakier-GingrasL, FerbeyreG, 2019. Ribosomal proteins control tumor suppressor pathways in response to nucleolar stress. Bioessays, 41(3):e1800183.

[22]LiFD, WangYC, LiY, et al., 2018. Quantitative analysis of the global proteome in peripheral blood mononuclear cells from patients with new-onset psoriasis. Proteomics, 18(19):e1800003.

[23]LinetteGP, LiY, RothK, et al., 1996. Cross talk between cell death and cell cycle progression: BCL-2 regulates NFAT-mediated activation. Proc Natl Acad Sci USA, 93(18):9545-9552.

[24]MisraS, SharmaS, AgarwalA, et al., 2010. Cell cycle-dependent regulation of the bi-directional overlapping promoter of human BRCA2/ZAR2 genes in breast cancer cells. Mol Cancer, 9:50.

[25]PandeyS, PatilS, BallavN, et al., 2020. Spatial targeting of Bcl-2 on endoplasmic reticulum and mitochondria in cancer cells by lipid nanoparticles. J Mater Chem B, 8(19):4259-4266.

[26]PihánP, Carreras-SuredaA, HetzC, 2017. BCL-2 family: integrating stress responses at the ER to control cell demise. Cell Death Differ, 24(9):1478-1487.

[27]PopgeorgievN, JabbourL, GilletG, 2018. Subcellular localization and dynamics of the Bcl-2 family of proteins. Front Cell Dev Biol, 6:13.

[28]QuinnLM, RichardsonH, 2004. Bcl-2 in cell cycle regulation. Cell Cycle, 3(1):6-8.

[29]RobinsonBH, LuoXP, PitkänenS, et al., 1998. Diagnosis of mitochondrial energy metabolism defects in tissue culture. Induction of MnSOD and bcl-2 in mitochondria from patients with complex I (NADH-CoQ reductase) deficiency. Biofactors, 7(3):229-230.

[30]SinghR, LetaiA, SarosiekK, 2019. Regulation of apoptosis in health and disease: the balancing act of BCL-2 family proteins. Nat Rev Mol Cell Biol, 20(3):175-193.

[31]TaitSWG, GreenDR, 2010. Mitochondria and cell death: outer membrane permeabilization and beyond. Nat Rev Mol Cell Biol, 11(9):621-632.

[32]TanakaS, SaitoK, ReedJC, 1993. Structure-function analysis of the Bcl-2 oncoprotein. Addition of a heterologous transmembrane domain to portions of the Bcl-2β protein restores function as a regulator of cell survival. J Biol Chem, 268(15):10920-10926.

[33]ThapaM, BommakantiA, ShamsuzzamanM, et al., 2013. Repressed synthesis of ribosomal proteins generates protein-specific cell cycle and morphological phenotypes. Mol Biol Cell, 24(23):3620-3633.

[34]TouzeauC, MaciagP, AmiotM, et al., 2018. Targeting Bcl-2 for the treatment of multiple myeloma. Leukemia, 32(9):1899-1907.

[35]VairoG, InnesKM, AdamsJM, 1996. Bcl-2 has a cell cycle inhibitory function separable from its enhancement of cell survival. Oncogene, 13(7):1511-1519.

[36]VairoG, SoosTJ, UptonTM, et al., 2000. Bcl-2 retards cell cycle entry through p27Kip1, pRB relative p130, and altered E2F regulation. Mol Cell Biol, 20(13):4745-4753.

[37]WangCH, WangLK, WuCC, et al., 2019. The ribosomal protein RPLP0 mediates PLAAT4-induced cell cycle arrest and cell apoptosis. Cell Biochem Biophys, 77(3):253-260.

[38]WangYQ, LiuYH, WangS, et al., 2020. Hydrogen agronomy: research progress and prospects. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 21(11):841-855.

[39]WarrenCFA, Wong-BrownMW, BowdenNA, 2019. BCL-2 family isoforms in apoptosis and cancer. Cell Death Dis, 10(3):177.

[40]XieMH, YenY, OwonikokoTK, et al., 2014. Bcl2 induces DNA replication stress by inhibiting ribonucleotide reductase. Cancer Res, 74(1):212-223.

[41]XieMH, ParkD, SicaGL, et al., 2020. Bcl2-induced DNA replication stress promotes lung carcinogenesis in response to space radiation. Carcinogenesis, 41(11):1565-1575.

[42]XuHD, QinZH, 2019. Beclin 1, Bcl-2 and autophagy. Adv Exp Med Biol, 1206:109-126.

[43]ZhouFF, YangY, XingD, 2011. Bcl-2 and Bcl-xL play important roles in the crosstalk between autophagy and apoptosis. FEBS J, 278(3):403-413.

Open peer comments: Debate/Discuss/Question/Opinion

<1>

Please provide your name, email address and a comment





Journal of Zhejiang University-SCIENCE, 38 Zheda Road, Hangzhou 310027, China
Tel: +86-571-87952783; E-mail: cjzhang@zju.edu.cn
Copyright © 2000 - 2024 Journal of Zhejiang University-SCIENCE