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2000
Volume 26, Issue 1
  • ISSN: 1871-5206
  • E-ISSN: 1875-5992

Abstract

Introduction

The presence of severe hypoxic stress can drive tumor growth, angiogenesis, and metastatic characteristics up-regulated hypoxia-inducible factor 1-alpha (HIF-1α). Hence, targeting HIF-1α is considered a promising strategy, as increased HIF-1α activity is a key factor in the aggressive phenotype of malignancies. In this study, we aimed to investigate the anti-cancer effects of several flavonoids, both single and in combination with PX-478, in breast cancer cell lines.

Methods

We tested the effects of luteolin and PX-478, both alone and in combination, on HIF-1α level in breast cancer cells under hypoxia using the cell viability assay. To determine the rationale for the cell growth inhibition induced by the luteolin+PX-478 combination, we conducted experiments to assess cell survival, apoptosis, cell cycle, invasion, and migration under both normoxic and hypoxic conditions. Furthermore, we evaluated the effect of this combination on DNA damage response under hypoxic stress Comet assay and immunofluorescence staining.

Results

Our findings revealed that the luteolin+PX-478 combination significantly suppressed the growth of MDA-MB-231 cells. In addition, we assessed time-dependent expression of HIF1α in MDA-MB-231 cells and observed that the combination of luteolin and PX-478 down-regulated the HIF-1α level. Finally, we found that the luteolin+PX-478 combination induced apoptosis and G cell cycle arrest and enhanced DNA damage response. This combination also sensitized breast cancer cells to ionizing radiation in hypoxic stress.

Discussion

The findings suggested that targeting HIF-1α with a combination of luteolin and PX-478 may provide a synergistic approach to suppressing tumor growth and enhancing therapeutic response under hypoxic conditions. The observed effects on apoptosis, cell cycle arrest, and DNA damage response indicated that this combination could be a promising strategy for overcoming hypoxia-induced resistance in breast cancer therapy.

Conclusion

Collectively, our results suggested the combination of luteolin and PX-478 to enhance the anti-cancer effects of PX-478 in breast carcinoma cells by impeding the cell growth and inducing DNA damage response under hypoxia.

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References

  1. BrayF. FerlayJ. SoerjomataramI. SiegelR.L. TorreL.A. JemalA. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries.CA Cancer J. Clin.201868639442410.3322/caac.21492 30207593
     [Google Scholar]
  2. EcclesS.A. WelchD.R. Metastasis: Recent discoveries and novel treatment strategies.Lancet200736995741742175710.1016/S0140‑6736(07)60781‑8 17512859
     [Google Scholar]
  3. ChaudaryN. HillR.P. Hypoxia and metastasis in breast cancer.Breast Dis.2007261556410.3233/BD‑2007‑26105 17473365
     [Google Scholar]
  4. KallinowskiF. ZanderR. HoeckelM. VaupelP. Tumor tissue oxygenation as evaluated by computerized-po2-histography.Int. J. Radiat. Oncol. Biol. Phys.199019495396110.1016/0360‑3016(90)90018‑F 2211264
     [Google Scholar]
  5. VaupelP. MayerA. BriestS. HöckelM. Hypoxia in breast cancer: Role of blood flow, oxygen diffusion distances, and anemia in the development of oxygen depletion.Adv. Exp. Med. Biol.200556633334210.1007/0‑387‑26206‑7_44 16594170
     [Google Scholar]
  6. HohenbergerP. FelgnerC. HaenschW. SchlagP.M. Tumor oxygenation correlates with molecular growth determinants in breast cancer.Breast Cancer Res. Treat.19984829710610.1023/A:1005921513083 9596481
     [Google Scholar]
  7. SchitoL. ReyS. TafaniM. ZhangH. WongC.C. RussoA. RussoM.A. SemenzaG.L. Hypoxia-inducible factor 1-dependent expression of platelet-derived growth factor B promotes lymphatic metastasis of hypoxic breast cancer cells.Proc. Natl. Acad. Sci. USA20121094027072716
     [Google Scholar]
  8. GrahamK. UngerE. Overcoming tumor hypoxia as a barrier to radiotherapy, chemotherapy and immunotherapy in cancer treatment.Int. J. Nanomedicine2018136049605810.2147/IJN.S140462 30323592
     [Google Scholar]
  9. DenglerV.L. GalbraithM.D. EspinosaJ.M. Transcriptional regulation by hypoxia inducible factors.Crit. Rev. Biochem. Mol. Biol.201449111510.3109/10409238.2013.838205 24099156
     [Google Scholar]
  10. WangG.L. SemenzaG.L. Purification and characterization of hypoxia-inducible factor 1.J. Biol. Chem.199527031230123710.1074/jbc.270.3.1230 7836384
     [Google Scholar]
  11. BhattaraiD. XuX. LeeK. Hypoxia‐inducible factor‐1 (HIF‐1) inhibitors from the last decade (2007 to 2016): A “structure–activity relationship” perspective.Med. Res. Rev.20183841404144210.1002/med.21477 29278273
     [Google Scholar]
  12. ParczykJ. RuhnauJ. PelzC. SchillingM. WuH. PiaskowskiN.N. EickholtB. KühnH. DankerK. KleinA. Dichloroacetate and PX-478 exhibit strong synergistic effects in a various number of cancer cell lines.BMC Cancer202121148110.1186/s12885‑021‑08186‑9 33931028
     [Google Scholar]
  13. PalayoorS.T. MitchellJ.B. CernaD. DeGraffW. John-AryankalayilM. ColemanC.N. PX‐478, an inhibitor of hypoxia‐inducible factor‐1α, enhances radiosensitivity of prostate carcinoma cells.Int. J. Cancer2008123102430243710.1002/ijc.23807 18729192
     [Google Scholar]
  14. ZhuY. ZangY. ZhaoF. LiZ. ZhangJ. FangL. LiM. XingL. XuZ. YuJ. Inhibition of HIF-1α by PX-478 suppresses tumor growth of esophageal squamous cell cancer in vitro and in vivo.Am. J. Cancer Res.20177511981212
     [Google Scholar]
  15. KopustinskieneD.M. JakstasV. SavickasA. BernatonieneJ. Flavonoids as anticancer agents.Nutrients202012245710.3390/nu12020457 32059369
     [Google Scholar]
  16. FotsisT. PepperM.S. AktasE. BreitS. RaskuS. AdlercreutzH. WähäläK. MontesanoR. SchweigererL. Flavonoids, dietary-derived inhibitors of cell proliferation and in vitro angiogenesis.Cancer Res.1997571429162921 9230201
     [Google Scholar]
  17. AbotalebM. SamuelS. VargheseE. VargheseS. KubatkaP. LiskovaA. BüsselbergD. Flavonoids in cancer and apoptosis.Cancers20181112810.3390/cancers11010028 30597838
     [Google Scholar]
  18. DükelM. TavsanZ. KayaliH.A. Flavonoids regulate cell death-related cellular signaling via ROS in human colon cancer cells.Process Biochem.2021101112510.1016/j.procbio.2020.10.002
     [Google Scholar]
  19. LiskovaA. KoklesovaL. SamecM. SmejkalK. SamuelS.M. VargheseE. AbotalebM. BiringerK. KudelaE. DankoJ. ShakibaeiM. KwonT.K. BüsselbergD. KubatkaP. Flavonoids in cancer metastasis.Cancers2020126149810.3390/cancers12061498 32521759
     [Google Scholar]
  20. SinghR. AgarwalR. Natural flavonoids targeting deregulated cell cycle progression in cancer cells.Curr. Drug Targets20067334535410.2174/138945006776055004 16515531
     [Google Scholar]
  21. GaoK. HenningS. NiuY. YoussefianA. SeeramN. XuA. HeberD. The citrus flavonoid naringenin stimulates DNA repair in prostate cancer cells.J. Nutr. Biochem.2006172899510.1016/j.jnutbio.2005.05.009 16111881
     [Google Scholar]
  22. ProcházkováD. BoušováI. WilhelmováN. Antioxidant and prooxidant properties of flavonoids.Fitoterapia201182451352310.1016/j.fitote.2011.01.018 21277359
     [Google Scholar]
  23. SamecM. LiskovaA. KoklesovaL. MersakovaS. StrnadelJ. KajoK. PecM. ZhaiK. SmejkalK. MirzaeiS. HushmandiK. AshrafizadehM. SasoL. BrockmuellerA. ShakibaeiM. BüsselbergD. KubatkaP. Flavonoids targeting HIF-1: Implications on cancer metabolism.Cancers202113113010.3390/cancers13010130 33401572
     [Google Scholar]
  24. GürlerS.B. KirazY. BaranY. Chapter 21 - Flavonoids in cancer therapy: Current and future trendsBiodiversity and Biomedicine.Academic Press2020403440
     [Google Scholar]
  25. KikuchiH. YuanB. HuX. OkazakiM. Chemopreventive and anticancer activity of flavonoids and its possibility for clinical use by combining with conventional chemotherapeutic agents.Am. J. Cancer Res.20199815171535 31497340
     [Google Scholar]
  26. ChouT.C. Drug combination studies and their synergy quantification using the Chou-Talalay method.Cancer Res.201070244044610.1158/0008‑5472.CAN‑09‑1947 20068163
     [Google Scholar]
  27. DukelM. Combination of naringenin and epicatechin sensitizes colon carcinoma cells to anoikis via regulation of the epithelial–mesenchymal transition (EMT).Mol. Cell. Toxicol.202319118720310.1007/s13273‑022‑00317‑y
     [Google Scholar]
  28. LivakK.J. SchmittgenT.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)).Method. Methods200125440240810.1006/meth.2001.1262 11846609
     [Google Scholar]
  29. DukelM. FiskinK. Combination of PAKs inhibitors IPA-3 and PF-3758309 effectively suppresses colon carcinoma cell growth by perturbing DNA damage response.Int. J. Radiat. Biol.202399234035410.1080/09553002.2022.2110326 35939342
     [Google Scholar]
  30. ParkM.Y. KimY. HaS.E. KimH.H. BhosaleP.B. AbusaliyaA. JeongS.H. KimG.S. Function and application of flavonoids in the breast cancer.Int. J. Mol. Sci.20222314773210.3390/ijms23147732 35887080
     [Google Scholar]
  31. SalehiB. FokouP.V.T. Sharifi-RadM. ZuccaP. PezzaniR. MartinsN. Sharifi-RadJ. The therapeutic potential of naringenin: A review of clinical trials.Pharmaceuticals20191211110.3390/ph12010011 30634637
     [Google Scholar]
  32. VetrivelP. KimS.M. SaralammaV.V.G. HaS.E. KimE.H. MinT.S. KimG.S. Function of flavonoids on different types of programmed cell death and its mechanism: A review.J. Biomed. Res.201933636310.7555/JBR.33.20180126
     [Google Scholar]
  33. DükelM. StreitfeldW.S. TangT.C.C. BackmanL.R.F. AiL. MayW.S. BrownK.D. The breast cancer tumor suppressor TRIM29 is expressed via ATM-dependent signaling in response to hypoxia.J. Biol. Chem.201629141215412155210.1074/jbc.M116.730960 27535224
     [Google Scholar]
  34. JingX. YangF. ShaoC. WeiK. XieM. ShenH. ShuY. Role of hypoxia in cancer therapy by regulating the tumor microenvironment.Mol. Cancer201918115710.1186/s12943‑019‑1089‑9 31711497
     [Google Scholar]
  35. McAleeseC.E. ChoudhuryC. ButcherN.J. MinchinR.F. Hypoxia-mediated drug resistance in breast cancers.Cancer Lett.202150218919910.1016/j.canlet.2020.11.045 33278499
     [Google Scholar]
  36. YongL. TangS. YuH. ZhangH. ZhangY. WanY. CaiF. The role of hypoxia-inducible factor-1 alpha in multidrug-resistant breast cancer.Front. Oncol.20221296493410.3389/fonc.2022.964934 36003773
     [Google Scholar]
  37. TangW. ZhaoG. Small molecules targeting HIF-1α pathway for cancer therapy in recent years.Bioorg. Med. Chem.202028211523510.1016/j.bmc.2019.115235 31843464
     [Google Scholar]
  38. MooringS.R. WangB. HIF-1 inhibitors as anti-cancer therapy.Sci. China Chem.2011541243010.1007/s11426‑010‑4187‑5
     [Google Scholar]
  39. JordanB.F. BlackK. RobeyI.F. RunquistM. PowisG. GilliesR.J. Metabolite changes in HT-29 xenograft tumors following HIF-1α inhibition with PX-478 as studied by MR spectroscopy in vivo and ex vivo.NMR Biomed.200518743043910.1002/nbm.977 16206237
     [Google Scholar]
  40. AvniR. CohenB. NeemanM. Hypoxic stress and cancer: Imaging the axis of evil in tumor metastasis.NMR Biomed.201124656958110.1002/nbm.1632 21793071
     [Google Scholar]
  41. Carcereri de PratiA. ButturiniE. RigoA. OppiciE. RossinM. BorieroD. MariottoS. Metastatic breast cancer cells enter into dormant state and express cancer stem cells phenotype under chronic hypoxia.J. Cell. Biochem.2017118103237324810.1002/jcb.25972 28262977
     [Google Scholar]
  42. DaiY. BaeK. SiemannD.W. Impact of hypoxia on the metastatic potential of human prostate cancer cells.Int. J. Radiat. Oncol. Biol. Phys.20118152152810.1016/j.ijrobp.2011.04.027
     [Google Scholar]
  43. CampbellK. CasanovaJ. A common framework for EMT and collective cell migration.Development2016143234291430010.1242/dev.139071 27899506
     [Google Scholar]
  44. YilmazM. ChristoforiG. EMT, the cytoskeleton, and cancer cell invasion.Cancer Metastasis Rev.2009281-2153310.1007/s10555‑008‑9169‑0 19169796
     [Google Scholar]
  45. PiresI.M. OlcinaM.M. AnbalaganS. PollardJ.R. ReaperP.M. CharltonP.A. McKennaW.G. HammondE.M. Targeting radiation-resistant hypoxic tumour cells through ATR inhibition.Br. J. Cancer2012107229129910.1038/bjc.2012.265 22713662
     [Google Scholar]
  46. SiegelR.L. MillerK.D. WagleN.S. JemalA. Cancer statistics, 2023.CA Cancer J. Clin.2023731174810.3322/caac.21763 36633525
     [Google Scholar]
  47. MuzB. de la PuenteP. AzabF. Kareem AzabA. The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy.Hypoxia20153839210.2147/HP.S93413
     [Google Scholar]
  48. InfantinoV. SantarsieroA. ConvertiniP. TodiscoS. IacobazziV. Cancer cell metabolism in hypoxia: Role of HIF-1 as key regulator and therapeutic target.Int. J. Mol. Sci.20212211570310.3390/ijms22115703 34071836
     [Google Scholar]
  49. RashidM. ZadehL.R. BaradaranB. MolaviO. GhesmatiZ. SabzichiM. RamezaniF. Up-down regulation of HIF-1α in cancer progression.Gene202179814579610.1016/j.gene.2021.145796 34175393
     [Google Scholar]
  50. BuiB.P. NguyenP.L. LeeK. ChoJ. Hypoxia-inducible factor-1: A novel therapeutic target for the management of cancer, drug resistance, and cancer-related pain.Cancers20221424605410.3390/cancers14246054 36551540
     [Google Scholar]
  51. TibesR. FalchookG.S. Von HoffD.D. WeissG.J. IyengarT. KurzrockR. Results from a phase I, dose-escalation study of PX-478, an orally available inhibitor of HIF-1α.J. Clin. Oncol.202428153076(Suppl.)
     [Google Scholar]
  52. ZhangY. LiH. ZhangJ. ZhaoC. LuS. QiaoJ. HanM. The combinatory effects of natural products and chemotherapy drugs and their mechanisms in breast cancer treatment.Phytochem. Rev.20201951179119710.1007/s11101‑019‑09628‑w
     [Google Scholar]
  53. JeonY.W. SuhY.J. Synergistic apoptotic effect of celecoxib and luteolin on breast cancer cells.Oncol. Rep.201329281982510.3892/or.2012.2158 23229294
     [Google Scholar]
  54. YangM.Y. WangC.J. ChenN.F. HoW.H. LuF.J. TsengT.H. Luteolin enhances paclitaxel-induced apoptosis in human breast cancer MDA-MB-231 cells by blocking STAT3.Chem. Biol. Interact.2014213606810.1016/j.cbi.2014.02.002 24525192
     [Google Scholar]
  55. ZhangL. LiuQ. HuangL. YangF. LiuA. ZhangJ. Combination of lapatinib and luteolin enhances the therapeutic efficacy of lapatinib on human breast cancer through the FOXO3a/NQO1 pathway.Biochem. Biophys. Res. Commun.2020531336437110.1016/j.bbrc.2020.07.049 32800546
     [Google Scholar]
  56. ZhaoT. RenH. JiaL. ChenJ. XinW. YanF. LiJ. WangX. GaoS. QianD. HuangC. HaoJ. Inhibition of HIF-1α by PX-478 enhances the anti-tumor effect of gemcitabine by inducing immunogenic cell death in pancreatic ductal adenocarcinoma.Oncotarget2015642250226210.18632/oncotarget.2948 25544770
     [Google Scholar]
  57. MuzB. WasdenK. AlhallakK. JeskeA. AzabF. SunJ. KingJ. KohnenD.R. VijR. AzabA.K. Inhibition of HIF-1a By PX-478 normalizes blood vessels, improves drug delivery and suppresses progression and dissemination in multiple myeloma.Blood20201363(Suppl. 1)10.1182/blood‑2020‑142154
     [Google Scholar]
  58. Panahi MeymandiA.R. AkbariB. SoltantoyehT. ShahosseiniZ. HosseiniM. HadjatiJ. MirzaeiH.R. PX-478, an HIF-1α inhibitor, impairs mesoCAR T cell antitumor function in cervical cancer.Front. Oncol.202414135780110.3389/fonc.2024.1357801 38425341
     [Google Scholar]
  59. XiaY. JiangL. ZhongT. The role of HIF-1α in chemo-/radioresistant tumors.OncoTargets Ther.2018113003301110.2147/OTT.S158206 29872312
     [Google Scholar]
  60. LangM. WangX. WangH. DongJ. LanC. HaoJ. HuangC. LiX. YuM. YangY. YangS. RenH. Arsenic trioxide plus PX-478 achieves effective treatment in pancreatic ductal adenocarcinoma.Cancer Lett.20163782879610.1016/j.canlet.2016.05.016 27212442
     [Google Scholar]
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Luteolin Enhances Anticancer Effects of PX-478 during Hypoxic Response in Metastatic Breast Cancer Cells
Anti-Cancer Agents in Medicinal Chemistry 26, 105 (2026); https://doi.org/10.2174/0118715206384227250901064037
/content/journals/acamc/10.2174/0118715206384227250901064037
/content/journals/acamc/10.2174/0118715206384227250901064037
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  • Article Type:     Research Article
Keyword(s): angiogenesis; breast cancer; HIF-1α; hypoxia; Luteolin; PX-478

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