Skip to main content

Advertisement

SpringerLink
Log in

Silibinin induced the apoptosis of Hep-2 cells via oxidative stress and down-regulating survivin expression

  • Laryngology
  • Published:
European Archives of Oto-Rhino-Laryngology Aims and scope Submit manuscript

Abstract

Silibinin is an anticancer and chemopreventive natural compound, which is extracted from milk thistle (Silybum marianum). It is reported that silibinin has anticancer efficacy in many malignant tumors. Laryngeal carcinoma is the second most common head and neck squamous carcinoma. In the present work, we investigated the effects of silibinin on laryngeal squamous cell carcinoma (LSCC) cell line Hep-2 cells. We found that silibinin induced the decrease of cell viability in Hep-2 cells with a concentration- and time-dependent manner. Moreover, silibinin resulted in the apoptosis of Hep-2 cells and had synergy effects with arsenic trioxide. Intracellular reactive oxygen species (ROS) accumulation increased because of silibinin exposure. ROS scavenger NAC alleviated the cytotoxicity of silibinin to Hep-2 cells. The mitochondrial membrane potential (MMP) was lost in Hep-2 cells treated with silibinin. Subsequently, silibinin induced the activation of caspase-3 in Hep-2 cells and caspase inhibitor Z-VAD-FMK inhibited the cytotoxicity of silibinin in Hep-2 cells. The survivin expression decreased after Hep-2 cells were treated with silibinin. In conclusion, silibinin induced the apoptosis of Hep-2 cells via oxidative stress and down-regulating survivin expression. Therefore, silibinin is a potential therapeutical agent against LSCC in future.

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
Fig. 7
Fig. 8

Similar content being viewed by others

Rererences

  1. Jemal A, Siegel R, Xu J, Ward E (2010) Cancer statistics. CA Cancer J Clin 60(5):277–300

    Article  PubMed  Google Scholar 

  2. Karatzanis AD, Psychogios G, Zenk J et al (2009) Comparison among different available surgical approaches in T1 glottic cancer. The Laryngoscope 119:1704–1708

    Article  PubMed  Google Scholar 

  3. Elsheikh MN, Rinaldo A, Hamakawa H et al (2006) Importance of molecular analysis in detecting cervical lymph node metastasis in head and neck squamous cell carcinoma. Head Neck 28:842–849

    Article  PubMed  Google Scholar 

  4. Gazak R, Walterova D, Kren V (2007) Silybin and silymarin—new and emerging applications in medicine. Curr Med Chem 14:315–338

    Article  PubMed  CAS  Google Scholar 

  5. Deep G, Agarwal R (2010) Antimetastatic efficacy of silibinin: molecular mechanisms and therapeutic potential against cancer. Cancer Metastasis Rev 29:447–463

    Article  PubMed  CAS  Google Scholar 

  6. Wu KJ, Zeng J, Zhu GD et al (2009) Silibinin inhibits prostate cancer invasion, motility and migration by suppressing vimentin and MMP-2 expression. Acta Pharmacol Sin 30:1162–1168

    Article  PubMed  CAS  Google Scholar 

  7. Kim S, Choi JH, Lim HI et al (2009) Silibinin prevents TPA-induced MMP-9 expression and VEGF secretion by inactivation of the Raf/MEK/ERK pathway in MCF-7 human breast cancer cells. Phytomedicine 16:573–580

    Article  PubMed  CAS  Google Scholar 

  8. Li L, Gao Y, Zhang L, Zeng J, He D, Sun Y (2008) Silibinin inhibits cell growth and induces apoptosis by caspase activation, down-regulating survivin and blocking EGFR-ERK activation in renal cell carcinoma. Cancer Lett 272:61–69

    Article  PubMed  CAS  Google Scholar 

  9. Singh RP, Tyagi A, Sharma G, Mohan S, Agarwal R (2008) Oral silibinin inhibits in vivo human bladder tumor xenograft growth involving down-regulation of survivin. Clin Cancer Res 14:300–308

    Article  PubMed  CAS  Google Scholar 

  10. Bang CI, Paik SY, Sun DI, Joo YH, Kim MS (2008) Cell growth inhibition and down-regulation of survivin by silibinin in a laryngeal squamous cell carcinoma cell line. Ann Otol Rhinol Laryngol 117:781–785

    PubMed  Google Scholar 

  11. Kong Q, Lillehei KO (1998) Antioxidant inhibitors for cancer therapy. Med Hypotheses 51:405–409

    Article  PubMed  CAS  Google Scholar 

  12. Duan W, Jin X, Li Q, Tashiro S, Onodera S, Ikejima T (2010) Silibinin induced autophagic and apoptotic cell death in HT1080 cells through a reactive oxygen species pathway. J Pharmacol Sci 113:48–56

    Article  PubMed  CAS  Google Scholar 

  13. Fan S, Li L, Chen S et al (2011) Silibinin induced-autophagic and apoptotic death is associated with an increase in reactive oxygen and nitrogen species in HeLa cells. Free Radic Res 45:1307–1324

    Article  PubMed  CAS  Google Scholar 

  14. Jiang YY, Huang H, Wang HJ et al (2011) Interruption of mitochondrial complex IV activity and cytochrome c expression activated O(2). (-)-mediated cell survival in silibinin-treated human melanoma A375–S2 cells via IGF-1R-PI3K-Akt and IGF-1R-PLC gamma-PKC pathways. Eur J Pharmacol 668:78–87

    Article  PubMed  CAS  Google Scholar 

  15. Noh EM, Yi MS, Youn HJ et al (2011) Silibinin enhances ultraviolet B-induced apoptosis in mcf-7 human breast cancer cells. J Breast Cancer 14:8–13

    Article  PubMed  Google Scholar 

  16. Ryan BM, Konecny GE, Kahlert S et al (2006) Survivin expression in breast cancer predicts clinical outcome and is associated with HER2, VEGF, urokinase plasminogen activator and PAI-1. Ann Oncol 17:597–604

    Article  PubMed  CAS  Google Scholar 

  17. Sah NK, Khan Z, Khan GJ, Bisen PS (2006) Structural, functional and therapeutic biology of survivin. Cancer Lett 244:164–171

    Article  PubMed  CAS  Google Scholar 

  18. Jha K, Shukla M, Pandey M (2012) Survivin expression and targeting in breast cancer. Surg Oncol 21:125–131

    Article  PubMed  Google Scholar 

  19. Wang Y, Kong W, Xiong X, Zhang S, Sun D (2005) [The expressions of survivin and VEGF in squamous cell carcinoma of larynx and the correlation between the two marks]. Lin chuang er bi yan hou ke za zhi. J Clin Otorhinolaryngol 19:838–841

    Google Scholar 

  20. Shen Z, Ren Y, Ye D, Guo J, Kang C, Ding H (2011) Significance and relationship between DJ-1 gene and surviving gene expression in laryngeal carcinoma. Eur J Histochem 55:e9

    Article  PubMed  CAS  Google Scholar 

  21. Cheng B, Yang X, Han Z, An L, Liu S (2008) Arsenic trioxide induced the apoptosis of laryngeal cancer via down-regulation of survivin mRNA. Auris Nasus Larynx 35:95–101

    Article  PubMed  Google Scholar 

  22. Kauntz H, Bousserouel S, Gosse F, Raul F (2011) Silibinin triggers apoptotic signaling pathways and autophagic survival response in human colon adenocarcinoma cells and their derived metastatic cells. Apoptosis 16:1042–1053

    Article  PubMed  CAS  Google Scholar 

  23. Cheung CW, Taylor PJ, Kirkpatrick CM et al (2007) Therapeutic value of orally administered silibinin in renal cell carcinoma: manipulation of insulin-like growth factor binding protein-3 levels. BJU Int 100:438–444

    Article  PubMed  CAS  Google Scholar 

  24. Kauntz H, Bousserouel S, Gosse F, Raul F (2012) The flavonolignan silibinin potentiates TRAIL-induced apoptosis in human colon adenocarcinoma and in derived TRAIL-resistant metastatic cells. Apoptosis 17:797–809

    Article  PubMed  CAS  Google Scholar 

  25. Son YG, Kim EH, Kim JY et al (2007) Silibinin sensitizes human glioma cells to TRAIL-mediated apoptosis via DR5 up-regulation and down-regulation of c-FLIP and survivin. Cancer Res 67:8274–8284

    Article  PubMed  CAS  Google Scholar 

  26. Dizaji MZ, Malehmir M, Ghavamzadeh A, Alimoghaddam K, Ghaffari SH (2012) Synergistic effects of arsenic trioxide and silibinin on apoptosis and invasion in human glioblastoma U87MG cell line. Neurochem Res 37:370–380

    Article  PubMed  CAS  Google Scholar 

  27. Flaig TW, Gustafson DL, Su LJ et al (2007) A phase I and pharmacokinetic study of silybin-phytosome in prostate cancer patients. Invest New Drugs 25:139–146

    Article  PubMed  CAS  Google Scholar 

  28. Pradhan SC, Girish C (2006) Hepatoprotective herbal drug, silymarin from experimental pharmacology to clinical medicine. Indian J Med Res 124:491–504

    PubMed  CAS  Google Scholar 

  29. Zhou B, Wu LJ, Li LH et al (2006) Silibinin protects against isoproterenol-induced rat cardiac myocyte injury through mitochondrial pathway after up-regulation of SIRT1. J Pharmacol Sci 102:387–395

    Article  PubMed  CAS  Google Scholar 

  30. Wang MJ, Lin WW, Chen HL et al (2002) Silymarin protects dopaminergic neurons against lipopolysaccharide-induced neurotoxicity by inhibiting microglia activation. Eur J Neurosci 16:2103–2112

    Article  PubMed  Google Scholar 

  31. Lu P, Mamiya T, Lu L et al (2010) Silibinin attenuates cognitive deficits and decreases of dopamine and serotonin induced by repeated methamphetamine treatment. Behav Brain Res 207:387–393

    Article  PubMed  CAS  Google Scholar 

  32. Lu P, Mamiya T, Lu LL et al (2009) Silibinin prevents amyloid beta peptide-induced memory impairment and oxidative stress in mice. Br J Pharmacol 157:1270–1277

    Article  PubMed  CAS  Google Scholar 

  33. Marrazzo G, Bosco P, La Delia F et al (2011) Neuroprotective effect of silibinin in diabetic mice. Neurosci Lett 504:252–256

    Article  PubMed  CAS  Google Scholar 

  34. Pallauf K, Rimbach G (2013) Autophagy, polyphenols and healthy ageing. Ageing Res Rev 12:237–252

    Article  PubMed  CAS  Google Scholar 

  35. Liu B, Yang P, Ye Y et al (2011) Role of ROS in the protective effect of silibinin on sodium nitroprusside-induced apoptosis in rat pheochromocytoma PC12 cells. Free Radic Res 45:835–847

    Article  PubMed  CAS  Google Scholar 

  36. Duan WJ, Li QS, Xia MY, Tashiro S, Onodera S, Ikejima T (2011) Silibinin activated p53 and induced autophagic death in human fibrosarcoma HT1080 cells via reactive oxygen species-p38 and c-Jun N-terminal kinase pathways. Biol Pharm Bull 34:47–53

    Article  PubMed  CAS  Google Scholar 

  37. Kim KW, Choi CH, Kim TH, Kwon CH, Woo JS, Kim YK (2009) Silibinin inhibits glioma cell proliferation via Ca2+/ROS/MAPK-dependent mechanism in vitro and glioma tumor growth in vivo. Neurochem Res 34:1479–1490

    Article  PubMed  CAS  Google Scholar 

  38. Zeng J, Sun Y, Wu K et al (2011) Chemopreventive and chemotherapeutic effects of intravesical silibinin against bladder cancer by acting on mitochondria. Mol Cancer Ther 10:104–116

    Article  PubMed  CAS  Google Scholar 

  39. Tiwari P, Kumar A, Balakrishnan S, Kushwaha HS, Mishra KP (2011) Silibinin-induced apoptosis in MCF7 and T47D human breast carcinoma cells involves caspase-8 activation and mitochondrial pathway. Cancer Invest 29:12–20

    Article  PubMed  CAS  Google Scholar 

  40. Ghobrial IM, Witzig TE, Adjei AA (2005) Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin 55:178–194

    Article  PubMed  Google Scholar 

  41. Agarwal C, Singh RP, Dhanalakshmi S et al (2003) Silibinin upregulates the expression of cyclin-dependent kinase inhibitors and causes cell cycle arrest and apoptosis in human colon carcinoma HT-29 cells. Oncogene 22:8271–8282

    Article  PubMed  CAS  Google Scholar 

  42. Tyagi A, Agarwal C, Harrison G, Glode LM, Agarwal R (2004) Silibinin causes cell cycle arrest and apoptosis in human bladder transitional cell carcinoma cells by regulating CDKI-CDK-cyclin cascade, and caspase 3 and PARP cleavages. Carcinogenesis 25:1711–1720

    Article  PubMed  CAS  Google Scholar 

  43. Kumar S (1999) Mechanisms mediating caspase activation in cell death. Cell Death Differ 6:1060–1066

    Article  PubMed  CAS  Google Scholar 

  44. Tyagi A, Singh RP, Agarwal C, Agarwal R (2006) Silibinin activates p53-caspase 2 pathway and causes caspase-mediated cleavage of Cip1/p21 in apoptosis induction in bladder transitional-cell papilloma RT4 cells: evidence for a regulatory loop between p53 and caspase 2. Carcinogenesis 27:2269–2280

    Article  PubMed  CAS  Google Scholar 

  45. Altieri DC (2008) Survivin, cancer networks and pathway-directed drug discovery. Nat Rev 8:61–70

    Article  CAS  Google Scholar 

  46. Zhao H, Ren J, Zhuo X, Ye H, Zou J, Liu S (2008) Prognostic significance of Survivin and CD44v6 in laryngeal cancer surgical margins. J Cancer Res Clin Oncol 134:1051–1058

    Article  PubMed  CAS  Google Scholar 

  47. Chen XM, Luan XY, Lei DP et al (2008) Suppression of survivin expression by short hairpin RNA induces apoptosis in human laryngeal carcinoma cells. ORL Oto-Rhino-Laryngol Relat Spec 70:168–175

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The present work was supported by Youth Fund of Jining Medical University (No. JYQ2011KZ005), Shandong Provincial Natural Science Foundation of China (ZR2010HL057, ZR2010HL045) and a Project of Shandong Province Higher Educational Science and Technology Program (No. J12LE10).

Conflict of interest

None declared.

Author information

Authors and Affiliations

  1. Jining Medical University, Jining, Shandong Province, People’s Republic China

    Xinxin Yang, Bo Bai & Jing Chen

  2. The Affiliated Hospital of Jining Medical University, Jining, Shandong Province, People’s Republic China

    Xiaoyu Li

  3. Rizhao People’s Hospital, Rizhao, Shandong Province, People’s Republic China

    Liangxiang An

Authors
  1. Xinxin Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaoyu Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Liangxiang An
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Bo Bai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jing Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Xinxin Yang.

Additional information

X. Yang, X. Li and L. An contributed equally.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yang, X., Li, X., An, L. et al. Silibinin induced the apoptosis of Hep-2 cells via oxidative stress and down-regulating survivin expression. Eur Arch Otorhinolaryngol 270, 2289–2297 (2013). https://doi.org/10.1007/s00405-013-2444-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00405-013-2444-x

Keywords

Search

Navigation