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Cellular Oncology

, Volume 38, Issue 3, pp 195–204 | Cite as

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

  • Zeynep Birsu Cincin
  • Miray Unlu
  • Bayram Kiran
  • Elif Sinem Bireller
  • Yusuf Baran
  • Bedia CakmakogluEmail author
Original Paper

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.

Keywords

Non-small cell lung cancer Hesperidin Anti-proliferative effect Apoptosis Gene expression profile 

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

Notes

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.

Supplementary material

13402_2015_222_Fig7_ESM.gif (33 kb)
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)
13402_2015_222_MOESM1_ESM.tif (1.7 mb)
Fig. 7 High Resolution Image (TIFF 1736 kb)
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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)
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ESM 1 (DOCX 14 kb)
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Copyright information

© International Society for Cellular Oncology 2015

Authors and Affiliations

  • Zeynep Birsu Cincin
    • 1
  • Miray Unlu
    • 2
  • Bayram Kiran
    • 3
  • Elif Sinem Bireller
    • 1
  • Yusuf Baran
    • 2
  • Bedia Cakmakoglu
    • 1
    Email author
  1. 1.Institute of Experimental Medical Research, Department of Molecular MedicineIstanbul UniversityCapaTurkey
  2. 2.Department of Molecular Biology and GeneticsIzmir Institute of TechnologyUrlaTurkey
  3. 3.Department of BiologyKastamonu UniversityKastamonuTurkey

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