Mechanism of the splicing factor 3B subunit 1/forkhead box protein M1 signaling pathway regulating cell cycle and intervening against the progression of cervical cancer

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Mechanism of the splicing factor 3B subunit 1/forkhead box protein M1 signaling pathway regulating cell cycle and intervening against the progression of cervical cancer

Basic research | 更新时间：2025-10-21

- Mechanism of the splicing factor 3B subunit 1/forkhead box protein M1 signaling pathway regulating cell cycle and intervening against the progression of cervical cancer
- Journal of Chongqing Medical University Vol. 49, Issue 9, Pages: 1129-1136(2024)
- 作者机构：
  
  湖北民族大学附属民大医院妇科，恩施 445000
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.003576
  CLC： R737.33
- Received：20 September 2023，
  
  Revised：2023-11-17，
  
  Accepted：15 April 2024，
  
  Published：28 September 2024
- 稿件说明：
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杨美平,邓颂,陈明倩,等.SF3B1/FOXM1信号通路调控细胞周期干预宫颈癌进展的机制研究[J].重庆医科大学学报,2024,49(9):1129-1136.

Yang Meiping,Deng Song,Chen Mingqian,et al.Mechanism of the splicing factor 3B subunit 1/forkhead box protein M1 signaling pathway regulating cell cycle and intervening against the progression of cervical cancer[J].Journal of Chongqing Medical University,2024,49(9):1129-1136.
杨美平,邓颂,陈明倩,等.SF3B1/FOXM1信号通路调控细胞周期干预宫颈癌进展的机制研究[J].重庆医科大学学报,2024,49(9):1129-1136. DOI： 10.13406/j.cnki.cyxb.003576.

Yang Meiping,Deng Song,Chen Mingqian,et al.Mechanism of the splicing factor 3B subunit 1/forkhead box protein M1 signaling pathway regulating cell cycle and intervening against the progression of cervical cancer[J].Journal of Chongqing Medical University,2024,49(9):1129-1136. DOI： 10.13406/j.cnki.cyxb.003576.

摘要

目的

2

基于剪接因子3B亚单位1（splicing-factor-3B-subunit-1，SF3B1）/叉头框蛋白M1（Forkhead Box M1，FOXM1）轴探讨调控细胞周期宫颈癌进展的作用机制。

方法

2

通过转染SF3B1过表达质粒（SF3B1 overexpression plasmid，pSF3B1）和2个SF3B1特异性短发夹RNA（short hairpin RNA）（shSF3B1-1、shSF3B1-2）过表达HeLa细胞或敲低SiHa细胞SF3B1。通过试验和乙炔基-2′-脱氧尿苷分析细胞的增殖活力和增殖率。流式细胞术分析细胞周期分布。应用细胞周期聚合酶链式反应（polymerase chain reaction，PCR）阵列分析、流式细胞术、染色质免疫共沉淀-定量聚合酶链式反应（chromatin immunoprecipitation -quantitative polymerase chain reaction，ChIP-qPCR）和荧光素酶实验明确SF3B1和FOXM1的靶向关系。

结果

2

SF3B1过表达后，HeLa细胞的生长率和增殖能力均显著增加（

P

<

0.05）。此外，SF3B1敲低后，SiHa细胞的生长率和增殖能力均显著降低（

P

<

0.05）。在HeLa细胞中，与pENTER组相比，pSF3B1组细胞在细胞周期的G

0

/G

1

期降低。然而，在SiHa细胞系中，敲低SF3B1提高了G

0

/G

1

期比率。此外，SF3B1过表达促进了HeLa细胞中G

1

期相关蛋白细胞周期蛋白D1表达，而敲低SF3B1抑制了SiHa细胞中周期蛋白D1表达。上调SF3B1表达明显增强FOXM1的表达，而下调SF3B1导致FOXM1的明显抑制。ChIP-qPCR分析进一步显示SF3B1在HeLa细胞系中与FOXM1启动子位点结合。在野生型FOXM1启动子中，与shNC组相比，荧光素酶活性在SF3B1敲低组中受到抑制（1.30±0.08 vs. 0.73±0.02、0.70±0.04，均

P

<

0.01），突变型FOXM1启动子不能在SiHa细胞中引发对shSF3B1的应答。HeLa细胞的增殖能力通过SF3B1过表达

而增强（shNC+pENTER vs. shNC+pSF3B1：25.1±1.9 vs. 61.2±3.8，

P

<

0.01），并被shFOXM1降低（shNC+pSF3B1 vs. shFOXM1+pSF3B1：61.2±3.8 vs. 18.5±1.9，

P

<

0.001），并且SF3B1过表达诱导的细胞G0/G1周期降低通过敲低FOXM1而部分逆转（shNC+pSF3B1 vs. shFOXM1+pSF3B1：35.0±1.5 vs. 58.0±1.8，

P

<

0.05）。

结论

2

SF3B1可以作为宫颈癌中的潜在肿瘤促进基因，其可能通过上调FOXM1的表达来促进宫颈癌细胞增殖并扰乱细胞周期。

Abstract

Objective

2

To investigate the mechanism of cell cycle regulation in intervening against the progression of cervical cancer based on the splicing factor 3B subunit 1（SF3B1）/forkhead box protein M1（FOXM1） axis.

Methods

2

SF3B1 overexpression in HeLa cells or SF3B1 knockdown in SiHa cells was achieved by transfection of SF3B1 overexpression plasmid（pSF3B1） and two SF3B1-specific short hairpin RNA（shSF3B1-1 and shSF3B1-2）. CCK-8 test and ethynyl-2′-deoxyuridine（EdU） were used to measure cell proliferation activity and proliferation rate；flow cytometry was used to observe cell cycle distribution；cell cycle PCR array，flow cytometry，chromatin co-immunoprecipitation-quantitative polymerase chain reaction（ChIP-qPCR），and luciferase experiments were used to clarify the targeting relationship between SF3B1 and FOXM1.

Results

2

After SF3B1 overexpression，there were significant increases in the growth rate and proliferation ability of HeLa cells（

P

<

0.05）. In addition，there were significant reductions in the growth rate and proliferation ability of SiHa cells after SF3B1 knockdown（

P

<

0.05）. In HeLa cells，compared with the pENTER group，the pSF3B1 group had a significant reduction in the G

0

/G

1

ratio，while in SiHa cells，SF3B1 knockdown significantly increased the G

0

/G

1

ratio. Furthermore，SF3B1 overexpression significantly promoted the expression of the G

1

phase-related protein cyclin D1 in HeLa cells，while

SF3B1 knockdown significantly inhibited the expression of cyclin D1 in SiHa cells. Upregulation of SF3B1 significantly enhanced the expression of FOXM1，while downregulation of SF3B1 significantly inhibited FOXM1. ChIP-qPCR analysis showed the binding of SF3B1 to FOXM1 promoter site in HeLa cells. In the wild-type FOXM1 promoter，compared with the shNC group，the SF3B1 knockdown group had inhibition of luciferase activity（1.30±0.08 vs. 0.73±0.02 and 0.70±0.04，

P

<

0.01），and the mutant FOXM1 promoter could not trigger a response to shSF3B1 in SiHa cells. The proliferation ability of HeLa cells was enhanced by SF3B1 overexpression（shNC+pENTER vs. shNC+pSF3B1：25.1±1.9 vs. 61.2±3.8，

P

<

0.01） and was reduced by shFOXM1（shNC+pSF3B1 vs. shFOXM1+pSF3B1： 61.2±3.8 vs. 18.5±1.9，

P

<

0.001），and the reduction in G0/G1 cycle induced by SF3B1 overexpression was partially reversed by FOXM1 knockdown（shNC+pSF3B1 vs. shFOXM1+pSF3B1：35.0±1.5 vs. 58.0±1.8，

P

<

0.05）.

Conclusion

2

SF3B1 can be used as a tumor-promoting gene in cervical cancer，and it can promote the proliferation of cervical cancer cells and disrupt the cell cycle by upregulating the expression of FOXM1.

关键词

Keywords

references

姜玮奇，王　静，贺海威，等 . 基于生物信息学筛选与鉴定宫颈癌的生物标志物微小染色体维持蛋白2 [J ] . 第二军医大学学报， 2021 ， 42 （ 4 ）： 399 - 410 ．

Jiang WQ ， Wang J ， He HW ， et al . Screening and verifying biomarker mini-chromosome maintenance protein 2 for cervical cancer based on bioinformatics [J ] . Acad J Second Mil Med Univ ， 2021 ， 42 （ 4 ）： 399 - 410 ．

Pappa KI ， Daskalakis G ， Anagnou NP . Metabolic rewiring is associated with HPV-specific profiles in cervical cancer cell lines [J ] . Sci Rep ， 2021 ， 11 ： 17718 ．

Liu JJ ， Ho JY ， Lee JE ， et al . Genomic，transcriptomic，and viral integration profiles associated with recurrent/metastatic progression in high-risk human papillomavirus cervical carcinomas [J ] . Cancer Med ， 2020 ， 9 （ 21 ）： 8243 - 8257 ．

Monteleone S ， Fedorov DG ， Townsend-Nicholson A ， et al . Hotspot identification and drug design of protein-protein interaction modulators using the fragment molecular orbital method [J ] . J Chem Inf Model ， 2022 ， 62 （ 16 ）： 3784 - 3799 ．

Lau E ， Han Y ， Williams DR ， et al . Splice-junction-based mapping of alternative isoforms in the human proteome [J ] . Cell Rep ， 2019 ， 29 （ 11 ）： 3751 - 3765 .

Gumińska N ， Zakryś B ， Milanowski R . A new type of circular RNA derived from nonconventional introns in nuclear genes of euglenids [J ] . J Mol Biol ， 2021 ， 433 （ 3 ）： 166758 ．

Kim Guisbert KS ， Mossiah I ， Guisbert E . Titration of SF3B1 activity reveals distinct effects on the transcriptome and cell physiology [J ] . Int J Mol Sci ， 2020 ， 21 （ 24 ）： 9641 ．

Zhao B ， Li Z ， Qian R ， et al . Cancer-associated mutations in SF3B1 disrupt the interaction between SF3B1 and DDX42 [J ] . J Biochem ， 2022 ， 172 （ 2 ）： 117 - 126 ．

Zhang QJ ， Di CX ， Yan JF ， et al . Inhibition of SF3b1 by pladienolide B evokes cycle arrest，apoptosis induction and p73 splicing in human cervical carcinoma cells [J ] . Artif Cells Nanomed Biotechnol ， 2019 ， 47 （ 1 ）： 1273 - 1280 ．

Park E ， Jiang Y ， Hao LL ， et al . Genetic variation and microRNA targeting of A-to-I RNA editing fine tune human tissue transcriptomes [J ] . Genome Biol ， 2021 ， 22 （ 1 ）： 77 ．

Putowski M ， Giannopoulos K . Perspectives on precision medicine in chronic lymphocytic leukemia：targeting recurrent mutations-NOTCH1，SF3B1，MYD88，BIRC3 [J ] . J Clin Med ， 2021 ， 10 （ 16 ）： 3735 ．

López-Cánovas JL ， Del Rio-Moreno M ， García-Fernandez H ， et al . Splicing factor SF3B1 is overexpressed and implicated in the aggressiveness and survival of hepatocellular carcinoma [J ] . Cancer Lett ， 2021 ， 496 ： 72 - 83 ．

Wang ZX . Regulation of cell cycle progression by growth factor-induced cell signaling [J ] . Cells ， 2021 ， 10 （ 12 ）： 3327 ．

Wang Q ， Wu SQ ， Gu YN ， et al . RASAL2 regulates the cell cycle and cyclin D1 expression through PI3K/AKT signalling in prostate tumorigenesis [J ] . Cell Death Discov ， 2022 ， 8 （ 1 ）： 275 ．

Liu ZQ ， Li YF ， Li XG ， et al . Overexpression of YBX1 promotes pancreatic ductal adenocarcinoma growth via the GSK3B/cyclin D1/cyclin E1 pathway [J ] . Mol Ther Oncolytics ， 2020 ， 17 ： 21 - 30 ．

Hasanvand A . The role of AMPK-dependent pathways in cellular and molecular mechanisms of metformin：a new perspective for treatment and prevention of diseases [J ] . Inflammopharmacology ， 2022 ， 30 （ 3 ）： 775 - 788 ．

Dang FB ， Nie L ， Zhou J ， et al . Inhibition of CK1ε potentiates the therapeutic efficacy of CDK4/6 inhibitor in breast cancer [J ] . Nat Commun ， 2021 ， 12 （ 1 ）： 5386 ．

刘彦军，姚四清，李　毅 . miR-20b-5p靶向调控Cyclin D1对NSCLC细胞生物学行为的影响 [J ] . 山东医药， 2021 ， 61 （ 25 ）： 36 - 40，45 ．

Liu YJ ， Yao SQ ， Li Y . Effect of miR-20b-5p regulating Cyclin D1 on biological function of NSCLC cells [J ] . Shandong Med J ， 2021 ， 61 （ 25 ）： 36 - 40，45 ．

Li ZL ， Liu XJ ， Yu HZ ， et al . USP21 regulates Hippo signaling to promote radioresistance by deubiquitinating FOXM1 in cervical cancer [J ] . Hum Cell ， 2022 ， 35 （ 1 ）： 333 - 347 ．

Hong HQ ， Zhu H ， Zhao SJ ， et al . The novel circCLK3/miR-320a/FoxM1 axis promotes cervical cancer progression [J ] . Cell Death Dis ， 2019 ， 10 （ 12 ）： 950 ．

Ji F ， Lu Y ， Chen SY ， et al . IGF 2 BP 2 -modified circular RNA circARHGAP12 promotes cervical cancer progression by interacting m 6 A/FOXM1 manner [J ] . Cell Death Discov ， 2021 ， 7 （ 1 ）： 215 ．

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