|Table of Contents|

Mechanism of piperlongumine-induced ferroptosis in glioma cells

Journal Of Modern Oncology[ISSN:1672-4992/CN:61-1415/R]

Issue:
2024 22
Page:
4262-4272
Research Field:
Publishing date:

Info

Title:
Mechanism of piperlongumine-induced ferroptosis in glioma cells
Author(s):
QIU Jianting1; 2; SHI Ji3; GUO Tangjun3; PIAO Haozhe1
1.Liaoning University of Traditional Chinese Medicine，Liaoning Shenyang 110847,China;2.The People's Hospital of Liaoning Province,Liaoning Shenyang 110013,China；3.Dalian Medical University,Liaoning Dalian 116044,China.
Keywords:
PL; glioblastoma; ferroptosis; EZH2
PACS:
R730.264
DOI:
10.3969/j.issn.1672-4992.2024.22.008
Abstract:
Objective:To explore the proliferation inhibition effect of piperlongumine (PL) on glioblastoma (GBM) in vitro experiments and the main ways,molecular mechanisms of GBM cells death.Methods:Use the CCK-8 assay kit to detect the effect of PL on the cell viability of glioma LN229 and A172.Conduct a colony formation experiment to detect the inhibitory effect of PL on glioma cell proliferation.Observe mitochondrial structural changes through electron microscopy.Immunofluorescence Ki67 staining experiment was used to detect cell proliferation ability.Immunofluorescence experiment was used to detect 4-HNE levels.Detect GSH and MDA levels using corresponding assay kits.Flow cytometry was used to detect intracellular ROS levels.CCK-8 assay was used to detect the recovery effect of various cell death inhibitors on PL-inhibited glioma cell proliferation.Flow cytometry apoptosis assay kit was used to detect the recovery effect of Fer-1.Bioinformatics analysis was used to find target genes and molecular docking evaluation.WB was used to detect iron death-related proteins.RT-qPCR was used to detect mRNA levels.The overexpressing EZH2 gene was transfected.CHIP experiment and qPCR was used to detect the interaction between H3K27me3 protein and Keap1 DNA.Results:PL can inhibit the cell viability of glioma cells LN229 and A172 and inhibit the proliferation of glioma cells.Ultrastructure showed that mitochondria in cells shrunk,and the double membrane density increased.Intracellular GSH levels decreased,while ROS,MDA,and 4-HNE levels increased.Ferroptosis inhibitors Fer-1 and Lip-1 can restore the inhibitory effect of PL on the proliferation of glioma cells.The small molecule ligand of PL is stable in binding with target protein macromolecule EZH2.Ferroptosis-related protein PTGS2 levels increased,Nrf2,xCT,GPX4 levels decreased.PL target protein EZH2 decreased,and its downstream protein H3K27me3 decreased,Nrf2 upstream protein Keap1 increased.EZH2 and Nrf2 mRNA levels showed no significant changes,while Keap1 mRNA levels increase.After overexpressing EZH2,H3K27me3,Keap1,and Nrf2 levels showed corresponding changes.CHIP experiments showed that H3K27me3 protein interacted with Keap1 DNA.Conclusion:We found that PL induces ferroptosis in glioma cells through a new EZH2/H3K27me3/Keap1/Nrf2 pathway,potentially providing a promising drug for GBM treatment.

References:

［1］LOUIS DN,PERRY A,REIFENBERGER G,et al.The 2016 World Health Organization classification of tumors of the central nervous system:a summary［J］.Acta Neuropathol,2016,1316:803-820.
［2］OSTROM QT,GITTLEMAN H,LIAO P,et al.CBTRUS statistical report:primary brain and central nervous system tumors diagnosed in the United States in 2007-2011［J］.Neuro Oncol,2014,Suppl 4(Suppl 4):iv1-63.
［3］ LIU J,LI C,WANG Y,et al.Prognostic and predictive factors in elderly patients with glioblastoma:A single-center retrospective study［J］.Front Aging Neurosci,2022,13:777962.
［4］ ELSAKA R,KITAGWA JM,REFAAT T,et al.Impact of extended adjuvant temozolamide beyond 6 months in the management of glioblastoma patients［J］.Am J Clin Oncol,2023,46(3):101-106.
［5］ PAUN L,MOIRAGHI A,JANNELLI G,et al.From focused ultrasound tumor ablation to brain blood barrier opening for high grade glioma:A systematic review［J］.Cancers (Basel),2021,13:5614.
［6］ GU X,WAN G,CHEN N,et al.DGKz plays crucial roles in the proliferation and tumorigenicity of human glioblastoma［J］.Int J Biol Sci,2019,15:1872-1881.
［7］ GILLMAN R,FIELD MA,SCHMITZ U,et al.Identifying cancer driver genes in individual tumors［J］.Comput Struct Biotechnol J,2023,21:5028-5038.
［8］ MARUSYK A,JANISZEWSKA M,POLYAK K.Intratumor heterogeneity:TheRosetta stone of therapy resistance［J］.Cancer Cell,2020,37:471-484.
［9］ WANG H,CHENG Y,MAO C,et al.Emerging mechanisms and targeted therapy of ferroptosis in cancer［J］.Mol Ther,2021,29:2185-2208.
［10］ STOCKWELL BR,FRIEDMANN ANGELI JP,BAYIR H,et al.Ferroptosis:A regulated cell death nexus linking metabolism,redox biology,and disease［J］.Cell,2017,171:273-285.
［11］ VARYNSKYI B,SHICK JA.Hacking the lipidome:new ferroptosis strategies in cancer therapy［J］.Biomedicines,2024,12(3):541.
［12］ YANG WS,KIM KJ,GASCHLER MM,et al.Peroxidation of polyunsaturated fatty acids by lipoxygenases drives ferroptosis［J］.Proc Natl Acad Sci USA,2016,113:E4966-4975.
［13］ DIXON SJ.Ferroptosis:bug or feature［J］.Immunol Rev,2017,277:150-157.
［14］ LEMASTERS JJ.Evolution of voltage-dependent anion channel function:frommolecular sieve to governator to actuator of ferroptosis ［J］.Front Oncol,2017,7:303.
［15］ IVANOV SD,SEMENOV AL,KOVAN'KO EG,et al.Effects of iron ionsand iron chelation on the efficiency of experimental radiotherapy of animals withgliomas［J］.Bull Exp Biol Med,2015,158:800-803.
［16］ SHIMADA K,SKOUTA R,KAPLAN A,et al.Global survey of cell death mechanisms reveals metabolic regulation of ferroptosis［J］.Nat Chem Biol,2016,12:497-503.
［17］ ALIM I,CAULFIELD JT,CHEN Y,et al.Selenium drives a trans criptional adaptive program to block ferroptosis and treatstroke［J］.Cell,2019,177:1262-1279.
［18］ PUNZIANO C,TROMBETTI S,CESARO E,et al.Antioxidant systems as modulators of ferroptosis:focus on transcription factors［J］.Antioxidants (Basel),2024,13(3):298.
［19］AZAM S,PARK JY,KIM IS,et al.Piperine and its metabolite's pharmacology in neurodegenerative and neurological diseases［J］.Biomedicines,2022,10(1):154.
［20］ ZHANG XS.Study on the mechanism of piperlongumine in iron death of gastric cancer cells［J］.The Chinese Journal of Clinical Pharmacology,2020,36(10):1280-1283.
［21］ YAMAGUCHIT Y.Piperlongumine rapidly induces the death of human pancreatic cancer cells mainly through the induction of ferroptosis［J］.Int J Oncol,2018,52:1011-1012.
［22］ LIU JM.Piperlongumine selectively kills glioblastoma multiforme cells via reactive oxygen species accumulation dependent JNK and p38 activation［J］.Biovhemical and Biophysical Research Communications,2013,437:87-93.
［23］文远超,余云湖，付晓红,等.荜茇酰胺对胶质瘤细胞增殖的影响［J］.中国公共卫生,2017,33(3):468-470. WEN YC，YU YH,FU XH,et al.Impact of piperlongumine on glioma proliferation［J］.Chinese Journal of Public Health,2017,33(3):468-470.
［24］BISHT S,MAO Y,EASWARAN H.Epigenetic dynamics of aging and cancer development:current concepts from studies mapping aging and cancer epigenomes［J］.Curr Opin Oncol,2024,36(2):82-92.
［25］ ZHANG XS.Study on the mechanism of piperlongumine in iron death of gastric cancer cells［J］.The Chinese Journal of Clinical Pharmacology,2020,36(10):1280-1283.
［26］ 王宁,康华丽,薛金慧,等.铁死亡相关基因TFRC在胃癌组织中的表达及其与化疗药物敏感性的关系［J］.现代肿瘤医学，2023,31(22):4183-4189. WANG Ning,KANG Huali,XUE Jinhui,et al.Expression of iron death related gene TFRC in gastric cancer tissue and its relationship with chemotherapy drug sensitivity［J］.Modern Oncology,2023,31(22):4183-4189.
［27］ ALEXIOU GA,GEROGIANNI P,V ARTHOLOMATOS E,et al.Deferiproneenhances temozolomide cytotoxicity in glioma cells［J］.Cancer Invest,2016,10:489-495.
［28］ RAMADOSS T,WEIMER DS,MAYROVITZ HN.Topical iron chelator therapy:current status and future prospects［J］.Cureus,2023,15(10):e47720.
［29］ LI Y,ZHANG J,ZHANG K,et al,Scutellaria barbata inhibits hepatocellular carcinoma tumorigenicity by inducing ferroptosis of hepatocellular carcinoma cells［J］.Front Oncol,2022,12:693395.
［30］ FORCINA GC,DIXON SJ.GPX4 at the crossroads of lipid homeostasis and ferroptosis［J］.Proteomics,2019,19(18):e1800311.
［31］程峰,张庸,王祥,等.谷胱甘肽过氧化物酶GPX4在铁死亡中的作用与机制研究进展［J］.现代肿瘤医学，2021,29(07):1254-1258. CHENG Feng,ZHANG Yong,WANG Xiang,et al.Research progress on the role and mechanism of GPX4 in ferroptosis［J］.Modern Oncology,2021,29(07):1254-1258.
［32］IMAI H,MATSUOKA M,KUMAGAI T,et al.Lipid peroxidation-dependent cell death regulated by GPx4 and ferroptosis［J］.Curr Top Microbiol Immunol,2017,403:143-170.
［33］KONG N,CHEN X,FENG J,et al.Baicalin induces ferroptosis in bladder cancer cells by downregulating FTH1［J］.Acta Pharm Sin B,2021,11:4045-4054.
［34］ZHANG S,XIN W,ANDERSON GJ,et al.Double-edge sword roles of iron in driving energy production versus instigating ferroptosis［J］.Cell Death Dis,2022,13:40.
［35］JAKOBSEN S,NIELSEN CU.Exploring amino acid transporters as therapeutic targets for cancer:an examination of inhibitor structures,selectivity issues,and discovery approaches［J］.Pharmaceutics,2024,16(2):197.
［36］DODSON M,CASTRO-PORTUGUEZ R,ZHANG DD.NRF2 plays a critical role in mitigating lipid peroxidation and ferroptosis［J］.Redox Biol,2019,23:101107.
［37］ 高洁,杜润蕾.Keap1-NRF2在癌症中的激活与失调机制［J］.现代肿瘤医学，2021,29(22):4068-4072. GAO Jie,DU Runlei.Keap1-NRF2 activation and dysregulation mechanisms in cancer［J］.Modern Oncology,2021,29(22):4068-4072.
［38］ DONG M,LU J,XUE H,et al.Anthocyanins from lycium ruthenicum murray mitigate cadmium-induced oxidative stress and testicular toxicity by activating the Keap1/Nrf2 signaling pathway［J］.Pharmaceuticals (Basel),2024,17(3):322.
［39］ AWUAH WA,TOUFIK AR,YARLAGADDA R,et al.Exploring the role of Nrf2 signaling in glioblastoma multiforme［J］.Discov Oncol,2022,13(1):94.
［40］ ANDRES CMC,PEREZDELA LASTRA JM,BUSTAMANTE MUNGUIRA E,et al.Electrophilic compounds in the human diet and their role in the induction of the transcription factor NRF2［J］.Int J Mol Sci,2024,25(6):3521.
［41］ PASINI D,DI CROCE L.Emerging roles for polycomb proteins in cancer［J］.Curr Opin Genet Dev,2016,36:50-58.
［42］WM FAN T,ISLAM JMM,HIGASHI RM,et al.Metabolic reprogramming driven by EZH2 inhibition depends on cell-matrix interactions［J］.J Biol Chem,2024,300(1):105485.
［43］YIN X,YANG S,ZHANG M,et al.The role and prospect of JMJD3 in stemcells and cancer［J］.Biomed Pharmacother,2019,118:109384.
［44］ LIU GZ,XU XW,TAO SH,et al.HBx facilitates ferroptosis in acute liver failure via EZH2 mediated SLC7A11 suppression［J］.J Biomed Sci,2021,28:67.
［45］LI JN,GAO W,ZHAO ZH,et al.Ginsenoside Rg1 reduced microglial activation and mitochondrial dysfunction to alleviate depression-like behaviour via the GAS5/EZH2/SOCS3/NRF2 axis［J］.Molecular Neurobiology,2022,59:2855-2873.
［46］ BEZERRA DP,PESSOA C,DE MORAES MO,et al,Overview of the therapeutic potential of piplartine (piperlongumine)［J］.European Journal of Pharmaceutical Sciences,2013,48(3):453-463.

Memo

Memo:
辽宁省沈阳市科技计划项目（编号：20-205-4-003）
Last Update: 1900-01-01