The research on <italic style="font-style: italic">FBW</italic>7 gene enhances antitumor effect of paclitaxel on pancreatic cancer through GSDME-mediated pyroptosis

Yuming JIA; Zeng YE; Yanli DENG; Shengchao LI; Zhilei ZHANG; Chao WANG; Xiaowu XU; Yi QIN; Li PENG

doi:10.19401/j.cnki.1007-3639.2023.10.001

您当前的位置：

首页 >

文章列表页 >

The research on FBW7 gene enhances antitumor effect of paclitaxel on pancreatic cancer through GSDME-mediated pyroptosis

Article | 更新时间：2025-12-31

- The research on FBW7 gene enhances antitumor effect of paclitaxel on pancreatic cancer through GSDME-mediated pyroptosis
- China Oncology Vol. 33, Issue 10, Pages: 889-897(2023)
- 作者机构：
  
  1. 河北医科大学第四医院肝胆外科，河北石家庄 050000
  2. 复旦大学附属肿瘤医院胰腺外科，复旦大学胰腺肿瘤研究所，复旦大学上海医学院肿瘤学系，上海 200032
  3. 河北医科大学第四医院检验科，河北石家庄 050000
- 作者简介：
- 基金信息：
  
  National Natural Science Foundation of China(82173281)
- DOI：10.19401/j.cnki.1007-3639.2023.10.001
  CLC： R735.9
- Received：13 August 2023，
  
  Revised：2023-10-13，
  
  Published：30 October 2023
- 稿件说明：
移动端阅览
Yuming JIA, Zeng YE, Yanli DENG, et al. The research on FBW7 gene enhances antitumor effect of paclitaxel on pancreatic cancer through GSDME-mediated pyroptosis[J]. China Oncology, 2023, 33(10): 889-897.
DOI：

Yuming JIA, Zeng YE, Yanli DENG, et al. The research on FBW7 gene enhances antitumor effect of paclitaxel on pancreatic cancer through GSDME-mediated pyroptosis[J]. China Oncology, 2023, 33(10): 889-897. DOI： 10.19401/j.cnki.1007-3639.2023.10.001.

摘要

背景与目的：

胰腺癌是恶性程度极高的肿瘤，多数患者在就诊时已处于晚期，全身化疗是重要的治疗手段，但是化疗耐药给临床治疗带来很多困扰。紫杉醇作为常用的化疗药物可以诱导肿瘤细胞发生凋亡，含F-框WD重复域蛋白7（F-box and WD repeat domain containing 7，FBW7）基因是一种抑癌基因，该基因的功能丧失会导致肿瘤发生、发展。研究表明，

FBW

7基因具有促进肿瘤细胞凋亡和抑制肿瘤增殖的作用。此外，该基因还被证实可以促进凋亡和通过铁死亡方式增加化疗药物的效果。焦亡是一种由焦孔素（gasdermin，GSDM）蛋白介导的细胞程序性死亡模式，这种细胞死亡往往还伴有炎症反应。本研究旨在探讨

FBW

7基因是否可以通过焦亡促进紫杉醇发挥抗肿瘤作用。

方法：

采用慢病毒转染构建FBW7过表达的PANC-1细胞株，采用免疫组织化学法检测临床标本中

FBW

7和

GSDME

基因表达的相关性，采用实时荧光定量聚合酶链式反应（real-time fluorescent quantitative polymerase chain reaction，RTFQ-PCR）和蛋白质印迹法（Western blot）检测mRNA表达和蛋白质水平。光镜下观察细胞经紫杉醇处理后的形态变化，采用细胞计数试剂盒-8（cell counting kit-8，CCK-8）检测紫杉醇对细胞活力的影响，采用流式细胞术、乳酸脱氢酶（lactate dehydrogenase，LDH）释放实验检测紫杉醇对肿瘤细胞的影响。Western blot检测经紫杉醇处理后肿瘤细胞中caspase-3和GSDME的活化情况。

结果：

RTFQ-PCR和Western blot检测结果表明，

FBW

7基因过表达可增加GSDME的表达。免疫组织化学检测结果表明，在体内

FBW

7和

GSDME

基因表达呈正相关。流式细胞术和LDH释放实验检测结果显示，FBW7过表达的胰腺癌细胞系PANC-1系经过紫杉醇药物刺激后，细胞死亡的水平较空载体（empty vector，EV）组显著增加。光镜下可观察到具有焦亡表现的FBW7过表达的PANC-1细胞株显著多于EV组细胞。CCK-8实验结果显示，FBW7过表达的细胞株在紫杉醇处理后细胞活性显著低于EV组细胞。Western blot检测结果显示，紫杉醇处理胰腺癌细胞系PANC-1后，FBW7过表达细胞株的活化caspase-3（active-caspase-3）和GSDME-NT蛋白水平增加，证明其有更加明显的活化。

FBW

7基因可以通过caspase-3/GSDME信号转导通路诱导的细胞焦亡增加PANC-1细胞对紫杉醇的敏感性。

结论：

FBW7可以通过上调GSDME的表达，增加胰腺癌细胞对紫杉醇诱导的细胞焦亡从而增加其对紫杉醇的敏感性，这种作用可能是通过caspase-3/GSDME信号转导通路实现的。

Abstract

Background and purpose:

Pancreatic cancer is a highly malignant disease. Most patients are in advanced stage upon diagnosis. Systemic chemotherapy is an important treatment method

but the chemotherapy drug resistance to tumors brings many problems to clinical treatment. As a commonly used chemotherapy drug

paclitaxel can induce apoptosis in tumor cells. The

FBW

7 gene is a tumor suppres

sor gene

and the loss of its function can lead to tumor occurrence and progression. Research has shown that it has the effect of promoting tumor cell apoptosis and inhibiting tumor proliferation. In addition

this gene has been proven to promote apoptosis and ferroptosis

which increase the effect of chemotherapy drugs. Pyroptosis is a programmed cell death mode mediated by gasdermin (GSDM) protein

and this cell death is often accompanied by inflammatory reactions. This study aimed to investigate whether

FBW

7 gene can promote the anti-tumor effect of paclitaxel by increasing pyroptosis.

Methods:

A PANC-1 cell line overexpressing

FBW

7 was constructed using lentivirus transfection. The correlation between

FBW

7 and

GSDME

gene expressions was detected by immunohistochemistry in clinical samples

and the expression levels of mRNA and protein were detected by real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. We observed the morphological changes of cells treated with paclitaxel under light microscopy. Cell counting kit-8 (CCK-8) was used to detect the effect of paclitaxel on cell viability

and flow cytometry and lactate dehydrogenase (LDH) release assay were performed to detect the effect of paclitaxel on cell death. Western blot was used to detect caspase-3 and GSDME activation after paclitaxel treatment.

Results:

RTFQ-PCR and Western blot experiments showed that overexpression of

FBW

7 gene increased the expression of GSDME. Immunohistochemical staining of pathological sections of clinical patients also showed that the expressions of

FBW

7 and

GSDME

genes was positively correlated

in vivo

. Flow cytometry and LDH release experiments showed that the level of cell death in pancreatic cancer cell line PANC-1 overexpressing

FBW

7 was significantly increased compared with its empty vector (EV) cells after b

eing treated with paclitaxel. Under light microscopy

it was observed that the number of cells with pyroptosis was significantly higher in PANC-1 cell lines overexpressing

FBW

7 than in EV cells. The CCK-8 experiment results showed that the cell viability was significantly lower in

FBW

7 overexpressed cell lines than in EV cells after paclitaxel treatment. Western blot experiment results showed that after pancreatic cancer cell line PANC-1 was treated with paclitaxel

the protein expressions of active-caspase-3 and GSDME-NT in

FBW

7 overexpression cell lines increased

which proved that they had more obvious activation

indicating that the

FBW

7 gene can increase the sensitivity of PANC-1 cells to paclitaxel through caspase-3/GSDME signaling pathway induced cell pyroptosis.

Conclusion:

FBW

7 can increase the sensitivity of pancreatic cancer cells to paclitaxel by increasing the expression of GSDME

which is realized through caspase-3/GSDME pathway.

关键词

Keywords

references

SIEGEL R L , MILLER K D , FUCHS H E , et al. Cancer statistics, 2022 [J ] . CA Cancer J Clin , 2022 , 72 ( 1 ): 7 - 33 . DOI: 10.3322/caac.v72.1 http://doi.org/10.3322/caac.v72.1 https://acsjournals.onlinelibrary.wiley.com/toc/15424863/72/1 https://acsjournals.onlinelibrary.wiley.com/toc/15424863/72/1

HUANG Y , WANG J W , HUANG J , et al. Pyroptosis, a target for cancer treatment? [J ] . Apoptosis , 2022 , 27 ( 1/2 ): 1 - 13 . DOI: 10.1007/s10495-021-01703-y http://doi.org/10.1007/s10495-021-01703-y

FANG Y , TIAN S W , PAN Y T , et al. Pyroptosis: a new frontier in cancer [J ] . Biomed Pharmacother , 2020 , 121 : 109595 . DOI: 10.1016/j.biopha.2019.109595 http://doi.org/10.1016/j.biopha.2019.109595

AACHOUI Y , SAGULENKO V , MIAO E A , et al. Inflammasome-mediated pyroptotic and apoptotic cell death, and defense against infection [J ] . Curr Opin Microbiol , 2013 , 16 ( 3 ): 319 - 326 . DOI: 10.1016/j.mib.2013.04.004 http://doi.org/10.1016/j.mib.2013.04.004

JORGENSEN I , RAYAMAJHI M , MIAO E A . Programmed cell death as a defence against infection [J ] . Nat Rev Immunol , 2017 , 17 ( 3 ): 151 - 164 . DOI: 10.1038/nri.2016.147 http://doi.org/10.1038/nri.2016.147

WANG Y B , YIN B , LI D N , et al. GSDME mediates caspase-3-dependent pyroptosis in gastric cancer [J ] . Biochem Biophys Res Commun , 2018 , 495 ( 1 ): 1418 - 1425 . DOI: 10.1016/j.bbrc.2017.11.156 http://doi.org/10.1016/j.bbrc.2017.11.156 https://linkinghub.elsevier.com/retrieve/pii/S0006291X17323331 https://linkinghub.elsevier.com/retrieve/pii/S0006291X17323331

ZHANG C C , LI C G , WANG Y F , et al. Chemotherapeutic paclitaxel and cisplatin differentially induce pyroptosis in A549 lung cancer cells via caspase-3/GSDME activation [J ] . Apoptosis , 2019 , 24 ( 3/4 ): 312 - 325 . DOI: 10.1007/s10495-019-01515-1 http://doi.org/10.1007/s10495-019-01515-1

WANG H , RONG X Y , ZHAO G , et al. The microbial metabolite trimethylamine N-oxide promotes antitumor immunity in triple-negative breast cancer [J ] . Cell Metab , 2022 , 34 ( 4 ): 581 - 594 .e8. DOI: 10.1016/j.cmet.2022.02.010 http://doi.org/10.1016/j.cmet.2022.02.010

WANG Y P , GAO W Q , SHI X Y , et al. Chemotherapy drugs induce pyroptosis through caspase-3 cleavage of a gasdermin [J ] . Nature , 2017 , 547 ( 7661 ): 99 - 103 . DOI: 10.1038/nature22393 http://doi.org/10.1038/nature22393 https://www.nature.com/articles/nature22393 https://www.nature.com/articles/nature22393

ZHANG Z B , ZHANG Y , XIA S Y , et al. Gasdermin E suppresses tumour growth by activating anti-tumour immunity [J ] . Nature , 2020 , 579 ( 7799 ): 415 - 420 . DOI: 10.1038/s41586-020-2071-9 http://doi.org/10.1038/s41586-020-2071-9

GERVASONI J E Jr , HINDENBURG A A , VEZERIDIS M P , et al. An effective in vitro antitumor response against human pancreatic carcinoma with paclitaxel and daunorubicin by induction of both necrosis and apoptosis [J ] . Anticancer Res , 2004 , 24 ( 5A ): 2617 - 2626 .

DAVIS R J , WELCKER M , CLURMAN B E . Tumor suppression by the FBW7 ubiquitin ligase: mechanisms and opportunities [J ] . Cancer Cell , 2014 , 26 ( 4 ): 455 - 464 . DOI: 10.1016/j.ccell.2014.09.013 http://doi.org/10.1016/j.ccell.2014.09.013

YE Z , ZHUO Q F , HU Q S , et al. FBW7-NRA41-SCD1 axis synchronously regulates apoptosis and ferroptosis in pancreatic cancer cells [J ] . Redox Biol , 2021 , 38 : 101807 . DOI: 10.1016/j.redox.2020.101807 http://doi.org/10.1016/j.redox.2020.101807 https://linkinghub.elsevier.com/retrieve/pii/S2213231720310120 https://linkinghub.elsevier.com/retrieve/pii/S2213231720310120

HASSAN M , WATARI H , ABUALMAATY A , et al. Apoptosis and molecular targeting therapy in cancer [J ] . Biomed Res Int , 2014 , 2014 : 1 - 23 .

SARHAN J , LIU B C , MUENDLEIN H I , et al. Caspase-8 induces cleavage of gasdermin D to elicit pyroptosis during Yersinia infection [J ] . Proc Natl Acad Sci U S A , 2018 , 115 ( 46 ): E10888 -E10897.

RÜHL S , SHKARINA K , DEMARCO B , et al. ESCRT-dependent membrane repair negatively regulates pyroptosis downstream of GSDMD activation [J ] . Science , 2018 , 362 ( 6417 ): 956 - 960 . DOI: 10.1126/science.aar7607 http://doi.org/10.1126/science.aar7607

ZHENG M , KARKI R , VOGEL P , et al. Caspase-6 is a key regulator of innate immunity, inflammasome activation, and host defense [J ] . Cell , 2020 , 181 ( 3 ): 674 - 687 .e13. DOI: S0092-8674(20)30333-0 http://doi.org/S0092-8674(20)30333-0

AKINO K , TOYOTA M , SUZUKI H , et al. Identification of DFNA5 as a target of epigenetic inactivation in gastric cancer [J ] . Cancer Sci , 2007 , 98 ( 1 ): 88 - 95 . DOI: 10.1111/cas.2007.98.issue-1 http://doi.org/10.1111/cas.2007.98.issue-1 https://onlinelibrary.wiley.com/toc/13497006/98/1 https://onlinelibrary.wiley.com/toc/13497006/98/1

KIM M S , CHANG X , YAMASHITA K , et al. Aberrant promoter methylation and tumor suppressive activity of the DFNA 5 gene in colorectal carcinoma [J ] . Oncogene , 2008 , 27 ( 25 ): 3624 - 3634 . DOI: 10.1038/sj.onc.1211021 http://doi.org/10.1038/sj.onc.1211021

CROES L , BEYENS M , FRANSEN E , et al. Large-scale analysis of DFNA 5 methylation reveals its potential as biomarker for breast cancer [J ] . Clin Epigenetics , 2018 , 10 : 51 . DOI: 10.1186/s13148-018-0479-y http://doi.org/10.1186/s13148-018-0479-y

HOECK J D , JANDKE A , BLAKE S M , et al. FBW7 controls neural stem cell differentiation and progenitor apoptosis via Notch and c-Jun [J ] . Nat Neurosci , 2010 , 13 ( 11 ): 1365 - 1372 . DOI: 10.1038/nn.2644 http://doi.org/10.1038/nn.2644

WANG Q Y , WANG Y P , DING J J , et al. A bioorthogonal system reveals antitumour immune function of pyroptosis [J ] . Nature , 2020 , 579 ( 7799 ): 421 - 426 . DOI: 10.1038/s41586-020-2079-1 http://doi.org/10.1038/s41586-020-2079-1

Views

2467

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Effect of liposome binding antisense oligonucleotide BP1003 on albumin-bound paclitaxel sensitivity in pancreatic cancer cells by inhibiting STAT3

A research on the mechanism of SERPINA3 promoting malignant progression and gemcitabine resistance of pancreatic cancer by inhibiting ferroptosis

New advances in basic research, clinical diagnosis and treatment of pancreatic cancer in 2024

An exploratory study of INPP4B, a biomarker of gemcitabine chemoresistance in pancreatic cancer

Research on uPAR promoting proliferation, migration, and chemoresistance of pancreatic cancer by inhibiting autophagy via MAPK signaling

Related Author

Hua FU

Guochao ZHOU

Rongmin CAI

Xin SONG

Dinghua YANG

Yuan HE

Juncheng GUO

Zhibin YE

Related Institution

Department of Hepatology and Gallbladder, Xiangxi Autonomous Prefecture People’s Hospital, Xiangxi

Department of Gastrointestial Surgery, Hebei General Hospital

Department of Gastrointestial Surgery, Heping Hospital Affiliated to Changzhi Medical College

The First Clinical College of Changzhi Medical College

Department of Pancreatic Surgery, Fudan University Shanghai Cancer Center, Department of Oncology, Shanghai Medical College, Fudan University, Shanghai Pancreatic Cancer Institute, Shanghai Key Laboratory of Precision Medicine for Pancreatic Cancer, Pancreatic Cancer Institute, Fudan University

AI问答

⁰

The research on FBW7 gene enhances antitumor effect of paclitaxel on pancreatic cancer through GSDME-mediated pyroptosis

DOI：10.19401/j.cnki.1007-3639.2023.10.001

摘要

Abstract

关键词

Keywords

references