Skip to main content

Advertisement

SpringerLink
Log in

Anti-proliferative, apoptotic and signal transduction effects of hesperidin in non-small cell lung cancer cells

  • Original Paper
  • Published:
Cellular Oncology Aims and scope Submit manuscript

Abstract

Purpose

Hesperidin, a glycoside flavonoid, is thought to act as an anti-cancer agent, since it has been found to exhibit both pro-apoptotic and anti-proliferative effects in several cancer cell types. The mechanisms underlying hesperidin-induced growth arrest and apoptosis are, however, not well understood. Here, we aimed to investigate the anti-proliferative and apoptotic effects of hesperidin on non-small cell lung cancer (NSCLC) cells and to investigate the mechanisms involved.

Methods

The anti-proliferative and apoptotic effects of hesperidin on two NSCLC-derived cell lines, A549 and NCI-H358, were determined using a WST-1 colorimetric assay, a LDH cytotoxicity assay, a Cell Death Detection assay, an AnnexinV-FITC assay, a caspase-3 assay and a JC-1 assay, respectively, all in a time- and dose-dependent manner. As a control, non-cancerous MRC-5 lung fibroblasts were included. Changes in whole genome gene expression profiles were assessed using an Illumina Human HT-12v4 beadchip microarray platform, and subsequent data analyses were performed using an Illumina Genome Studio and Ingenuity Pathway Analyser (IPA).

Results

We found that after hesperidin treatment, A549 and NCI-H358 cells exhibited decreasing cell proliferation and increasing caspase-3 and other apoptosis-related activities, in conjunction with decreasing mitochondrial membrane potential activities, in a dose- and time-dependent manner. Through a GO analysis, by which changes in gene expression profiles were compared, we found that the FGF and NF-κB signal transduction pathways were most significantly affected in the hesperidin treated NCI-H358 and A549 NSCLC cells.

Conclusions

Our results indicate that hesperidin elicits an in vitro growth inhibitory effect on NSCLC cells by modulating immune response-related pathways that affect apoptosis. When confirmed in vivo, hesperidin may serve as a novel anti-proliferative agent for non-small cell lung 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

Abbreviations

EF:

Nucleosomal enrichment factor

FBS:

Fetal bovine serum

FGF:

Fibroblast growth factor

FITC:

Fluorescein ısothiocyanate

IPA:

Ingenuity pathway analysis

LDH:

Lactate dehydrogenase

MEM-α:

Eagle’s minimum essential medium

NF-kB:

Nuclear factor kappa B

NSCLC:

Non-small cell lung cancer

PS:

Phosphatidylserine

RPMI-1640:

Roswell Park Memorial Institute-1640

References

  1. W. Pao, N. Girard, New driver mutations in non-small-cell lung cancer. Lancet Oncol. 12, 175–180 (2011)

    Article  CAS  PubMed  Google Scholar 

  2. W.D. Travis, Classification of lung cancer. Semin. Roentgenol. 46, 178–186 (2011)

    Article  PubMed  Google Scholar 

  3. Q. Wu, Y.F. Chen, J. Fu, Q.H. You, S.M. Wang, X. Huang, X.J. Feng, S.H. Zhang, Short hairpin RNA-mediated down-regulation of CENP-A attenuates the aggressive phenotype of lung adenocarcinoma cells. Cell. Oncol. 37, 399–407 (2014)

    Article  CAS  Google Scholar 

  4. A. Koren, H. Motaln, T. Cufer, Lung cancer stem cells: a biological and clinical perspective. Cell. Oncol. 36, 265–275 (2013)

    Article  CAS  Google Scholar 

  5. N. Peled, M.W. Wynes, N. Ikeda, T. Ohira, K. Yoshida, J. Qian, M. Ilouze, R. Brenner, Y. Kato, C. Mascaux, F.R. Hirsch, Insulin-like growth factor-1 receptor (IGF-1R) as a biomarker for resistance to the tyrosine kinase inhibitor gefitinib in non-small cell lung cancer. Cell. Oncol. 36, 277–288 (2013)

    Article  CAS  Google Scholar 

  6. A. Maier, AL. Peille, V. Vuaroqueaux, M. Lahn. Anti-tumor activity of the TGF-β receptor kinase inhibitor galunisertib (LY2157299 monohydrate) in patient-derived tumor xenografts. Cell. Oncol. 2015 Jan 9. [Epub ahead of print] DOI 10.1007/s13402-014-0210-8

  7. P. Ulivi, R. Silvestrini, Role of quantitative and qualitative characteristics of free circulating DNA in the management of patients with non-small cell lung cancer. Cell. Oncol. 36, 439–448 (2013)

    Article  CAS  Google Scholar 

  8. G. Giaccone, Twenty-five years of treating advanced NSCLC: what have we achieved? Ann. Oncol. 15 Suppl 4: iv81–83 (2004)

  9. J.W. Neal, L.V. Sequist, Exciting new targets in lung cancer therapy: ALK, IGF-1R, HDAC, and Hh. Curr. Treat. Options in Oncol. 11, 36–44 (2010)

    Article  Google Scholar 

  10. R. Sangha, P.N. Lara, P.C. Mack, D.R. Gandara, Beyond antiepidermal growth factor receptors and antiangiogenesis strategies for nonsmall cell lung cancer: exploring a new frontier. Curr. Opin. Oncol. 21, 116–123 (2009)

    Article  CAS  PubMed  Google Scholar 

  11. S. Mateen, K. Raina, R. Agarwal, Chemopreventive and anti-cancer efficacy of silibinin against growth and progression of lung cancer. Nutr. Cancer J. 65, 3–11 (2013)

    Article  CAS  Google Scholar 

  12. J. Nones, T.C.E. Spohr, F.C. Gomes, Hesperidin, a flavone glycoside, as mediator of neuronal survival. Neurochem. Res. 36, 1776–1784 (2011)

    Article  CAS  PubMed  Google Scholar 

  13. T. Tanaka, R. Takahashi, Flavonoids and asthma. Nutrients 10, 2128–2143 (2013)

    Article  Google Scholar 

  14. J.A. Manthey, K. Grohmann, N. Guthrie, Biological properties of citrus flavonoids pertaining to cancer and inflammation. Curr. Med. Chem. 8, 135–153 (2001)

    Article  CAS  PubMed  Google Scholar 

  15. J. Yu, L. Wang, R.L. Walzem, E.G. Miller, L.M. Pike, B.S. Patil, Antioxidant activity of citrus limonoids, flavonoids, and coumarins. J. Agric. Food Chem. 53, 2009–2014 (2005)

    Article  CAS  PubMed  Google Scholar 

  16. S.L. Hwang, P.H. Shih, G.C. Yen, Neuroprotective effects of citrus flavonoids. J. Agric. Food Chem. 60, 877–885 (2012)

    Article  CAS  PubMed  Google Scholar 

  17. E. Meiyanto, A. Hermawan, Anindyajati: natural products for cancer-targeted therapy: citrus flavonoids as potent chemopreventive agents. Asian. Pac. J. Cancer Prev. 13, 427–1436 (2012)

    Article  PubMed  Google Scholar 

  18. J.A. Manthey, N. Guthrie, Antiproliferative activities of citrus flavonoids against six human cancer cell lines. J. Agric. Food Chem. 50, 5837–5843 (2002)

    Article  CAS  PubMed  Google Scholar 

  19. G. Saiprasad, P. Chitra, R. Manikandan, G. Sudhandiran, Hesperidin alleviates oxidative stress and downregulates the expressions of proliferative and inflammatory markers in azoxymethane-induced experimental colon carcinogenesis in mice. Inflamm. Res. 62, 425–440 (2013)

    Article  CAS  PubMed  Google Scholar 

  20. O.G. Benavente, J. Castillo, M. Alcaraz, V. Vicente, J.A. Del, A. Ortuno, Beneficial action of Citrus flavonoids on multiple cancer-related biological pathways. Curr. Cancer Drug Targets 7, 795–809 (2007)

    Article  Google Scholar 

  21. J. Nones, T.C.E. Spohr, F.C. Gomes, Hesperidin, a flavone glycoside, as mediator of neuronal survival. Neurochem. Res. 36, 1776–1784 (2011)

    Article  CAS  PubMed  Google Scholar 

  22. V. Gaur, A. Kumar, Hesperidin pre-treatment attenuates NO-mediated cerebral ischemic reperfusion injury and memory dysfunction. Pharmacol. Rep. 62, 635–648 (2010)

    Article  CAS  PubMed  Google Scholar 

  23. S. Ou, Pharmacological action of hesperidin. Zhong. Yao. Cai. 25, 531–533 (2002)

    PubMed  Google Scholar 

  24. O. Benavente-Garcia, J. Castillo, Update on uses and properties of citrus flavonoids: new findings in anticancer, cardiovascular, and anti-inflammatory activity. J. Agric. Food Chem. 56, 6185–6205 (2008)

    Article  CAS  PubMed  Google Scholar 

  25. A. Chanet, D. Milenkovic, C. Manach, A. Mazur, C. Morand, Citrus flavanones: what is their role in cardiovascular protection? J. Agric. Food Chem. 60, 8809–8822 (2012)

    Article  CAS  PubMed  Google Scholar 

  26. B.D. Sahu, M. Kuncha, G.J. Sindhura, R. Sistla, Hesperidin attenuates cisplatin-induced acute renal injury by decreasing oxidative stress, inflammation and DNA damage. Phytomedicine 20, 453–460 (2013)

    Article  CAS  PubMed  Google Scholar 

  27. J.R. Patil, K.N. Chidambara Murthy, G.K. Jayaprakasha, M.B. Chetti, B.S. Patil, Bioactive compounds from Mexican lime [Citrus aurantifolia] juice induce apoptosis in human pancreatic cells. J. Agric. Food Chem 57, 10933–10942 (2009)

    Article  CAS  PubMed  Google Scholar 

  28. M. Galleano, O. Pechanova, C.G. Fraga, Hypertension, nitric oxide, oxidants, and dietary plant polyphenols. Curr. Pharm. Biotechnol. 11, 837–848 (2010)

    Article  CAS  PubMed  Google Scholar 

  29. H.J. Park, M.J. Kim, E. Ha, J.H. Chung, Apoptotic effect of hesperidin through caspase3 activation in human colon cancer cells, SNU-C4. Phytomedicine 15, 147–151 (2008)

    Article  CAS  PubMed  Google Scholar 

  30. A. Ghorban, M. Nazari, M. Jeddi-Tehrani, H. Zand, The citrus flavonoid hesperidin induces p53 and inhibits NF-κB activation in order to trigger apoptosis in NALM-6 cells: involvement of PPARγ-dependent mechanism. Eur. J. Nutr. 51, 39–46 (2012)

    Article  Google Scholar 

  31. M. Nazari, A. Ghorbani, A. Hekmat-Doost, M. Jeddi-Tehrani, H. Zand, Inactivation of nuclear factor-κB by citrus flavanone hesperidin contributes to apoptosis and chemo-sensitizing effect in Ramos cells. Eur. J. Pharmacol. 650, 526–533 (2011)

    Article  CAS  PubMed  Google Scholar 

  32. K. Mohankumar, S. Pajaniradje, S. Sridharan, V.K. Singh, L. Ronsard, A.C. Banerjea, B.C. Selvanesan, M.S. Coumar, L. Periyasamy, R. Rajagopalan, Apoptosis induction by an analog of curcumin (BDMC-A) in human laryngeal carcinoma cells through intrinsic and extrinsic pathways. Cell. Oncol. 37, 439–454 (2014)

    Article  CAS  Google Scholar 

  33. S. Aranganathan, N. Nalini, Efficacy of the potential chemopreventive agent, hesperetin [citrus flavanone], on 1,2-dimethylhydrazine induced colon carcinogenesis. Food Chem. Toxicol. 47, 2594–2600 (2009)

    Article  CAS  PubMed  Google Scholar 

  34. C.J. Lee, L. Wilson, M.A. Jordan, V. Nguyen, J. Tang, G. Smiyun, Hesperidin suppressed proliferations of both human breast cancer and androgen-dependent prostate cancer cells. Phytother. Res. Suppl 1, S15–S19 (2010)

    Article  Google Scholar 

  35. G. Saiprasad, P. Chitra, R. Manikandan, G. Sudhandiran, Hesperidin induces apoptosis and triggers autophagic markers through inhibition of Aurora-A mediated phosphoinositide-3-kinase/Akt/mammalian target of rapamycin and glycogen synthase kinase-3 beta signalling cascades in experimental colon carcinogenesis. Eur. J. Cancer 50, 2489–2507 (2014)

    Article  CAS  PubMed  Google Scholar 

  36. S. Yumnam, HS. Park, MK. Kim, A. Nagappan, GE. et al, Hesperidin induces paraptosis like cell death in hepatoblatoma, HepG2 cells: involvement of ERK1/2 MAPK. PLoS One 30;9(6):e101321 (2014)

Download references

Acknowledgments

This work was funded by Istanbul University Scientific Research Project number 9205. We would like to thank Mr. David Chapman for editing the manuscript.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Institute of Experimental Medical Research, Department of Molecular Medicine, Istanbul University, Capa, Istanbul, Turkey

    Zeynep Birsu Cincin, Elif Sinem Bireller & Bedia Cakmakoglu

  2. Department of Molecular Biology and Genetics, Izmir Institute of Technology, Urla, Izmir, Turkey

    Miray Unlu & Yusuf Baran

  3. Department of Biology, Kastamonu University, Kastamonu, Turkey

    Bayram Kiran

Authors
  1. Zeynep Birsu Cincin
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Miray Unlu
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Bayram Kiran
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Elif Sinem Bireller
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yusuf Baran
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bedia Cakmakoglu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bedia Cakmakoglu.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplemental Fig. 7

FGF signaling network. FGF network of 50 μM hesperidin-stimulated genes in A549 (a), NCI-H358(b) and MRC-5(c) cells. Ingenuity pathways analysis (IPA) software was used to identify genes involved in the FGF signaling that were differentially expressed in A549, NCI-H358 and MRC-5 cells. Genes labeled in red and green were identified as up- and down-regulated, respectively, whereas other genes were identified on the basis of the network analysis. (GIF 32 kb)

Fig. 7

High Resolution Image (TIFF 1736 kb)

Supplemental Fig. 8

NF-κB signaling network. NF-κB network of 50 μM hesperidin-stimulated genes in NCI-H358 cells. Ingenuity pathways analysis (IPA) software was used to identify genes involved in the NF-κB signaling that were differentially expressed in NCI-H358 cells. Genes labeled in red and green were identified as up- and down-regulated, respectively, whereas other genes were identified on the basis of the network analysis. (JPEG 131 kb)

ESM 1

(DOCX 14 kb)

ESM 2

(DOCX 14 kb)

ESM 3

(DOC 31 kb)

ESM 4

(DOCX 15 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Birsu Cincin, Z., Unlu, M., Kiran, B. et al. Anti-proliferative, apoptotic and signal transduction effects of hesperidin in non-small cell lung cancer cells. Cell Oncol. 38, 195–204 (2015). https://doi.org/10.1007/s13402-015-0222-z

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13402-015-0222-z

Keywords

Search

Navigation