Skip to main content

Advertisement

SpringerLink
Log in

Synergistic effect of fisetin combined with sorafenib in human cervical cancer HeLa cells through activation of death receptor-5 mediated caspase-8/caspase-3 and the mitochondria-dependent apoptotic pathway

  • Original Article
  • Published:
Tumor Biology

Abstract

Combining antitumor agents with bioactive compounds is a potential strategy for improving the effect of chemotherapy on cancer cells. The goal of this study was to elucidate the antitumor effect of the flavonoid, fisetin, combined with the multikinase inhibitor, sorafenib, against human cervical cancer cells in vitro and in vivo. The combination of fisetin and sorafenib synergistically induced apoptosis in HeLa cells, which is accompanied by a marked increase in loss of mitochondrial membrane potential. Apoptosis induction was achieved by caspase-3 and caspase-8 activation which increased the ratio of Bax/Bcl-2 and caused the subsequent cleavage of PARP level while disrupting the mitochondrial membrane potential in HeLa cells. Decreased Bax/Bcl-2 ratio level and mitochondrial membrane potential were also observed in siDR5-treated HeLa cells. In addition, in vivo studies revealed that the combined fisetin and sorafenib treatment was clearly superior to sorafenib treatment alone using a HeLa xenograft model. Our study showed that the combination of fisetin and sorafenib exerted better synergistic effects in vitro and in vivo than either agent used alone against human cervical cancer, and this synergism was based on apoptotic potential through a mitochondrial- and DR5-dependent caspase-8/caspase-3 signaling pathway. This combined fisetin and sorafenib treatment represents a novel therapeutic strategy for further clinical developments in advanced cervical cancer.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Saslow D, Solomon D, Lawson HW, Killackey M, Kulasingam SL, et al. American Cancer Society, American Society for Colposcopy and Cervical Pathology, and American Society for Clinical Pathology screening guidelines for the prevention and early detection of cervical cancer. J Low Genit Tract Dis. 2012;16:175–204.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Kamangar F, Dores GM, Anderson WF. Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol. 2006;24:2137–50.

    Article  PubMed  Google Scholar 

  3. del Campo JM, Prat A, Gil-Moreno A, Perez J, Parera M. Update on novel therapeutic agents for cervical cancer. Gynecol Oncol. 2008;110:S72–6.

    Article  PubMed  Google Scholar 

  4. Chen HX, Cleck JN. Adverse effects of anticancer agents that target the VEGF pathway. Nat Rev Clin Oncol. 2009;6:465–77.

    Article  CAS  PubMed  Google Scholar 

  5. Thorburn A. Death receptor-induced cell killing. Cell Signal. 2004;16:139–44.

    Article  CAS  PubMed  Google Scholar 

  6. Riedl SJ, Salvesen GS. The apoptosome: signalling platform of cell death. Nat Rev Mol Cell Biol. 2007;8:405–13.

    Article  CAS  PubMed  Google Scholar 

  7. Babbitt SE, Sutherland MC, Francisco BS, Mendez DL, Kranz RG. Mitochondrial cytochrome c biogenesis: no longer an enigma. Trends Biochem Sci. 2015;40:446–55.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Rong Y, Distelhorst CW. Bcl-2 protein family members: versatile regulators of calcium signaling in cell survival and apoptosis. Annu Rev Physiol. 2008;70:73–91.

    Article  CAS  PubMed  Google Scholar 

  9. Murakami Y, Miller JW, Vavvas DG. RIP kinase-mediated necrosis as an alternative mechanisms of photoreceptor death. Oncotarget. 2011;2:497–509.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Maduro JH, Noordhuis MG, ten Hoor KA, Pras E, Arts HJ, et al. The prognostic value of TRAIL and its death receptors in cervical cancer. Int J Radiat Oncol Biol Phys. 2009;75:203–11.

    Article  CAS  PubMed  Google Scholar 

  11. Duenas-Gonzalez A, Cetina L, Mariscal I, de la Garza J. Modern management of locally advanced cervical carcinoma. Cancer Treat Rev. 2003;29:389–99.

    Article  PubMed  Google Scholar 

  12. Moon YJ, Wang X, Morris ME. Dietary flavonoids: effects on xenobiotic and carcinogen metabolism. Toxicol In Vitro. 2006;20:187–210.

    Article  CAS  PubMed  Google Scholar 

  13. Chen YC, Shen SC, Lee WR, Lin HY, Ko CH, et al. Wogonin and fisetin induction of apoptosis through activation of caspase 3 cascade and alternative expression of p21 protein in hepatocellular carcinoma cells SK-HEP-1. Arch Toxicol. 2002;76:351–9.

    Article  CAS  PubMed  Google Scholar 

  14. Liao YC, Shih YW, Chao CH, Lee XY, Chiang TA. Involvement of the ERK signaling pathway in fisetin reduces invasion and migration in the human lung cancer cell line A549. J Agric Food Chem. 2009;57:8933–41.

    Article  CAS  PubMed  Google Scholar 

  15. Kang KA, Piao MJ, Hyun JW. Fisetin induces apoptosis in human nonsmall lung cancer cells via a mitochondria-mediated pathway. In Vitro Cell Dev Biol Anim. 2015;51:300–9.

    Article  CAS  PubMed  Google Scholar 

  16. Khan N, Adhami VM, Mukhtar H. Apoptosis by dietary agents for prevention and treatment of prostate cancer. Endocr Relat Cancer. 2010;17:R39–52.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Chou RH, Hsieh SC, Yu YL, Huang MH, Huang YC, et al. Fisetin inhibits migration and invasion of human cervical cancer cells by down-regulating urokinase plasminogen activator expression through suppressing the p38 MAPK-dependent NF-kappaB signaling pathway. PLoS One. 2013;8, e71983.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Ying TH, Yang SF, Tsai SJ, Hsieh SC, Huang YC, et al. Fisetin induces apoptosis in human cervical cancer HeLa cells through ERK1/2-mediated activation of caspase-8-/caspase-3-dependent pathway. Arch Toxicol. 2012;86:263–73.

    Article  CAS  PubMed  Google Scholar 

  19. Bagi CM, Gebhard DF, Andresen CJ. Antitumor effect of vascular endothelial growth factor inhibitor sunitinib in preclinical models of hepatocellular carcinoma. Eur J Gastroenterol Hepatol. 2012;24:563–74.

    Article  CAS  PubMed  Google Scholar 

  20. Cheng AL, Kang YK, Chen Z, Tsao CJ, Qin S, et al. Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma: a phase III randomised, double-blind, placebo-controlled trial. Lancet Oncol. 2009;10:25–34.

    Article  CAS  PubMed  Google Scholar 

  21. Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, et al. Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 2008;359:378–90.

    Article  CAS  PubMed  Google Scholar 

  22. Amantini C, Morelli MB, Santoni M, Soriani A, Cardinali C, et al. Sorafenib induces cathepsin B-mediated apoptosis of bladder cancer cells by regulating the Akt/PTEN pathway. The Akt inhibitor, perifosine, enhances the sorafenib-induced cytotoxicity against bladder cancer cells. Oncoscience. 2015;2:395–409.

    Article  PubMed  PubMed Central  Google Scholar 

  23. Broecker-Preuss M, Muller S, Britten M, Worm K, Schmid KW, et al. Sorafenib inhibits intracellular signaling pathways and induces cell cycle arrest and cell death in thyroid carcinoma cells irrespective of histological origin or BRAF mutational status. BMC Cancer. 2015;15:184.

    Article  PubMed  PubMed Central  Google Scholar 

  24. Ishijima N, Kanki K, Shimizu H, Shiota G. Activation of AMP-activated protein kinase by retinoic acid sensitizes hepatocellular carcinoma cells to apoptosis induced by sorafenib. Cancer Sci. 2015;106:567–75.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Hsieh SC, Tsai JP, Yang SF, Tang MJ, Hsieh YH. Metformin inhibits the invasion of human hepatocellular carcinoma cells and enhances the chemosensitivity to sorafenib through a downregulation of the ERK/JNK-mediated NF-kappaB-dependent pathway that reduces uPA and MMP-9 expression. Amino Acids. 2014;46:2809–22.

    Article  CAS  PubMed  Google Scholar 

  26. Teixeira SF, Alexandre de Azevedo R, Salomon MA, Jorge SD, Levy D, et al. Synergistic anti-tumor effects of the combination of a benzofuroxan derivate and sorafenib on NCI-H460 human large cell lung carcinoma cells. Biomed Pharmacother. 2014;68:1015–22.

    Article  CAS  PubMed  Google Scholar 

  27. Dornetshuber-Fleiss R, Heilos D, Mohr T, Richter L, Sussmuth RD, et al. The naturally born fusariotoxin enniatin B and sorafenib exert synergistic activity against cervical cancer in vitro and in vivo. Biochem Pharmacol. 2015;93:318–31.

    Article  CAS  PubMed  Google Scholar 

  28. Chen CM, Hsieh YH, Hwang JM, Jan HJ, Hsieh SC, et al. Fisetin suppresses ADAM9 expression and inhibits invasion of glioma cancer cells through increased phosphorylation of ERK1/2. Tumour Biol. 2015;36:3407–15.

    Article  CAS  PubMed  Google Scholar 

  29. Kotipatruni RP, Ren X, Thotala D, Jaboin JJ. NDRG4 is a novel oncogenic protein and p53 associated regulator of apoptosis in malignant meningioma cells. Oncotarget. 2015;6:17594–604.

    Article  PubMed  PubMed Central  Google Scholar 

  30. Tsai JP, Lee CH, Ying TH, Lin CL, Hsueh JT, et al. Licochalcone A induces autophagy through PI3K/Akt/mTOR inactivation and autophagy suppression enhances Licochalcone A-induced apoptosis of human cervical cancer cells. Oncotarget. 2015;6:28851–66.

    PubMed  PubMed Central  Google Scholar 

  31. Aggarwal BB. Signalling pathways of the TNF superfamily: a double-edged sword. Nat Rev Immunol. 2003;3:745–56.

    Article  CAS  PubMed  Google Scholar 

  32. Chipuk JE, Bouchier-Hayes L, Green DR. Mitochondrial outer membrane permeabilization during apoptosis: the innocent bystander scenario. Cell Death Differ. 2006;13:1396–402.

    Article  CAS  PubMed  Google Scholar 

  33. Williams MM, Cook RS. Bcl-2 family proteins in breast development and cancer: could Mcl-1 targeting overcome therapeutic resistance? Oncotarget. 2015;6:3519–30.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Gross A, McDonnell JM, Korsmeyer SJ. BCL-2 family members and the mitochondria in apoptosis. Genes Dev. 1999;13:1899–911.

    Article  CAS  PubMed  Google Scholar 

  35. Lopez J, Tait SW. Mitochondrial apoptosis: killing cancer using the enemy within. Br J Cancer. 2015;112:957–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Ren W, Qiao Z, Wang H, Zhu L, Zhang L. Flavonoids: promising anticancer agents. Med Res Rev. 2003;23:519–34.

    Article  CAS  PubMed  Google Scholar 

  37. Syed DN, Lall RK, Chamcheu JC, Haidar O, Mukhtar H. Involvement of ER stress and activation of apoptotic pathways in fisetin induced cytotoxicity in human melanoma. Arch Biochem Biophys. 2014;563:108–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Tripathi R, Samadder T, Gupta S, Surolia A, Shaha C. Anticancer activity of a combination of cisplatin and fisetin in embryonal carcinoma cells and xenograft tumors. Mol Cancer Ther. 2011;10:255–68.

    Article  CAS  PubMed  Google Scholar 

  39. Wu MS, Lien GS, Shen SC, Yang LY, Chen YC. N-acetyl-L-cysteine enhances fisetin-induced cytotoxicity via induction of ROS-independent apoptosis in human colonic cancer cells. Mol Carcinog. 2014;53 Suppl 1:E119–29.

    Article  CAS  PubMed  Google Scholar 

  40. Kuo HC, Lee HJ, Hu CC, Shun HI, Tseng TH. Enhancement of esculetin on Taxol-induced apoptosis in human hepatoma HepG2 cells. Toxicol Appl Pharmacol. 2006;210:55–62.

    Article  CAS  PubMed  Google Scholar 

  41. Oikonomou E, Pintzas A. The TRAIL of oncogenes to apoptosis. Biofactors. 2013;39:343–54.

    Article  CAS  PubMed  Google Scholar 

  42. Kroemer G. The proto-oncogene Bcl-2 and its role in regulating apoptosis. Nat Med. 1997;3:614–20.

    Article  CAS  PubMed  Google Scholar 

  43. Youle RJ, Strasser A. The BCL-2 protein family: opposing activities that mediate cell death. Nat Rev Mol Cell Biol. 2008;9:47–59.

    Article  CAS  PubMed  Google Scholar 

  44. Dole M, Nunez G, Merchant AK, Maybaum J, Rode CK, et al. Bcl-2 inhibits chemotherapy-induced apoptosis in neuroblastoma. Cancer Res. 1994;54:3253–9.

    CAS  PubMed  Google Scholar 

  45. Chen JJ, Mikelis CM, Zhang Y, Gutkind JS, Zhang B. TRAIL induces apoptosis in oral squamous carcinoma cells—a crosstalk with oncogenic Ras regulated cell surface expression of death receptor 5. Oncotarget. 2013;4:206–17.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Ko H, Jeong MH, Jeon H, Sung GJ, So Y, et al. Delphinidin sensitizes prostate cancer cells to TRAIL-induced apoptosis, by inducing DR5 and causing caspase-mediated HDAC3 cleavage. Oncotarget. 2015;6:9970–84.

    Article  PubMed  PubMed Central  Google Scholar 

  47. Tanaka R, Tomosugi M, Horinaka M, Sowa Y, Sakai T. Metformin causes G1-phase arrest via down-regulation of MiR-221 and enhances TRAIL sensitivity through DR5 Up-regulation in pancreatic cancer cells. PLoS One. 2015;10, e0125779.

    Article  PubMed  PubMed Central  Google Scholar 

  48. Zhang Y, Ge Y, Chen Y, Li Q, Chen J, et al. Cellular and molecular mechanisms of silibinin induces cell-cycle arrest and apoptosis on HeLa cells. Cell Biochem Funct. 2012;30:243–8.

    Article  CAS  PubMed  Google Scholar 

  49. Bishayee K, Ghosh S, Mukherjee A, Sadhukhan R, Mondal J, et al. Quercetin induces cytochrome-c release and ROS accumulation to promote apoptosis and arrest the cell cycle in G2/M, in cervical carcinoma: signal cascade and drug-DNA interaction. Cell Prolif. 2013;46:153–63.

    Article  CAS  PubMed  Google Scholar 

  50. Haas NB, Manola J, Ky B, Flaherty KT, Uzzo RG, et al. Effects of adjuvant Sorafenib and Sunitinib on cardiac function in renal cell carcinoma patients without overt metastases: results from ASSURE, ECOG 2805. Clin Cancer Res. 2015;21:4048–54.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Pal HC, Baxter RD, Hunt KM, Agarwal J, Elmets CA, et al. Fisetin, a phytochemical, potentiates sorafenib-induced apoptosis and abrogates tumor growth in athymic nude mice implanted with BRAF-mutated melanoma cells. Oncotarget. 2015;6:28296–311.

    Article  PubMed  PubMed Central  Google Scholar 

  52. Pal HC, Diamond AC, Strickland LR, Kappes JC, Katiyar SK, et al.. Fisetin, a dietary flavonoid, augments the anti-invasive and anti-metastatic potential of sorafenib in melanoma. Oncotarget. 2015. doi:10.18632/oncotarget.6237

  53. Kiprianova I, Remy J, Milosch N, Mohrenz IV, Seifert V, et al. Sorafenib sensitizes glioma cells to the BH3 mimetic ABT-737 by targeting MCL1 in a STAT3-dependent manner. Neoplasia. 2015;17:564–73.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgments

This work was supported by grants from Chang Bing Show Chwan Memorial Hospital (RD104025).

Author information

Author notes

    Authors and Affiliations

    1. Department of Obstetrics and Gynecology, Chang Bing Show Chwan Memorial Hospital, Lugang Town, Changhua County, Taiwan

      Ming-Te Lin

    2. Liberal Arts Center, Da-Yeh University, Changhua, Taiwan

      Ming-Te Lin

    3. Institute of Biochemistry, Microbiology and Immunology, Chung Shan Medical University, Taichung, Taiwan

      Chia-Liang Lin, Chun-Wen Cheng, Chu-Liang Lin, Chih-Chien Wu & Yi-Hsien Hsieh

    4. Department of Biochemistry and Molecular Biology, College of Biological Sciences, University of California Davis, Davis, CA, USA

      Tzu-Yu Lin

    5. Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan

      Shun-Fa Yang

    6. Department of Biochemistry, School of Medicine, Chung Shan Medical University, Taichung, Taiwan

      Yi-Hsien Hsieh

    7. Clinical Laboratory, Chung Shan Medical University Hospital, Taichung, Taiwan

      Yi-Hsien Hsieh

    8. School of Medicine, Tzu Chi University, Hualien, Taiwan

      Jen-Pi Tsai

    9. Department of Internal Medicine, Division of Nephrology, Dalin Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Chiayi, Taiwan

      Jen-Pi Tsai

    Authors
    1. Ming-Te Lin
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Chia-Liang Lin
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Tzu-Yu Lin
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Chun-Wen Cheng
      View author publications

      You can also search for this author in PubMed Google Scholar

    5. Shun-Fa Yang
      View author publications

      You can also search for this author in PubMed Google Scholar

    6. Chu-Liang Lin
      View author publications

      You can also search for this author in PubMed Google Scholar

    7. Chih-Chien Wu
      View author publications

      You can also search for this author in PubMed Google Scholar

    8. Yi-Hsien Hsieh
      View author publications

      You can also search for this author in PubMed Google Scholar

    9. Jen-Pi Tsai
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding authors

    Correspondence to Yi-Hsien Hsieh or Jen-Pi Tsai.

    Ethics declarations

    Conflicts of interest

    None

    Additional information

    Ming-Te Lin and Chia-Liang Lin contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Lin, MT., Lin, CL., Lin, TY. et al. Synergistic effect of fisetin combined with sorafenib in human cervical cancer HeLa cells through activation of death receptor-5 mediated caspase-8/caspase-3 and the mitochondria-dependent apoptotic pathway. Tumor Biol. 37, 6987–6996 (2016). https://doi.org/10.1007/s13277-015-4526-4

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s13277-015-4526-4

    Keywords

    Search

    Navigation