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Pediatric Surgery International

, Volume 35, Issue 12, pp 1369–1378 | Cite as

Epidermal growth factor receptor/heme oxygenase-1 axis is involved in chemoresistance to cisplatin and pirarubicin in HepG2 cell lines and hepatoblastoma specimens

  • Takashi KobayashiEmail author
  • Masayuki Kubota
  • Yoshiaki Kinoshita
  • Yuki Arai
  • Toshiyuki Oyama
  • Naoki Yokota
  • Koichi Saito
  • Yasunobu Matsuda
  • Mami Osawa
Original Article
  • 66 Downloads

Abstract

Purpose

To investigate the possibility that the antioxidant stress protein Heme oxygenase-1 (HO-1) is involved in the acquisition of chemoresistance in cisplatin and pirarubicin (CITA) therapy.

Methods

Human hepatoblastoma-derived cell line (HepG2) was used to generate a knockdown cell line of HO-1 by small interfering RNA (siRNA). Expression of HO-1, epidermal growth factor receptor (EGFR), Akt, and extracellular signal-regulated kinase1/2 (ERK1/2) was examined by Western blot. The cytotoxic effect of cisplatin, pirarubicin, and EGFR inhibitor was examined by trypan blue staining. In human hepatoblastoma specimens (n = 5), changes of HO-1 expression were examined immunohistochemically before and after CITA therapy.

Results

HO-1 expression in HepG2 cells was increased by the treatment of cisplatin (CDDP) and pirarubicin (THP) dose-dependently. In HO-1 knockdown HepG2 cells, the HO-1 was not expressed and the percentage of trypan blue-positive cells (dead cells) was significantly increased after treatment of CDDP and THP. The EGFR inhibitor decreased the levels of HO-1, phospho-Akt and phospho-ERK1/2 in HepG2 cells. Combination treatment of EGFR inhibitor with CDDP and THP increased the cytotoxic effect in HepG2 cells. In human hepatoblastoma specimens, 4 of the 5 patients (80%) showed HO-1 expression changed much stronger in the viable tumor cells after CITA therapy.

Conclusion

The cytotoxic effects of CDDP and THP were both enhanced under HO-1 knockdown conditions as well as under conditions that inhibit the activation pathway of HO-1 by EGFR inhibitors. EGFR/HO-1 axis may be involved in acquiring chemoresistance in HepG2 cell lines as well as in human hepatoblastoma.

Keywords

Hepatoblastoma Cisplatin Pirarubicin Heme oxygenase-1 Epidermal growth factor receptor 

Notes

Acknowledgements

This study was supported in part by a Grant-in-Aid for Scientific Research (17H04270F).

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Takashi Kobayashi
    • 1
    Email author
  • Masayuki Kubota
    • 1
    • 4
  • Yoshiaki Kinoshita
    • 1
  • Yuki Arai
    • 1
  • Toshiyuki Oyama
    • 1
  • Naoki Yokota
    • 1
  • Koichi Saito
    • 1
  • Yasunobu Matsuda
    • 2
  • Mami Osawa
    • 3
  1. 1.Department of Pediatric SurgeryNiigata University Graduate School of Medical and Dental SciencesNiigataJapan
  2. 2.Department of Medical TechnologyNiigata University Graduate School of Health SciencesNiigataJapan
  3. 3.Division of Digestive and General SurgeryNiigata University Graduate School of Medical and Dental SciencesNiigataJapan
  4. 4.Kokuraminami Medical Care HospitalKitakyu-syuJapan

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