Histological and biochemical analysis of DNA damage after BNCT in rat model

Mitsuko Masutani, Diaz Baiseitov, Tasuku Itoh, Takahisa Hirai, Kulzhan Berikkhanova, Yasufumi Murakami, Zhaxybay Zhumadilov, Yoshio Imahori, Masaharu Hoshi, Jun Itami

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

To understand the mechanism of tumor cell death induced by boron neutron capture therapy (BNCT) and to optimize BNCT condition, we used rat tumor graft models and histological and biochemical analyses were carried out focusing on DNA damage response. Rat lymphosarcoma cells were grafted subcutaneously into male Wister rats. The rats with developed tumors were then treated with neutron beam irradiation 45min after injection of 330mg/kg bodyweight boronophenylalanine (10BPA) (+BPA) or saline control (-BPA). BNCT was carried out in the National Nuclear Center of the Republic of Kazakhstan (neutron flux: 1×109nvt/s, fluence: 6×1011nvt) with the presence of background γ-irradiation of 33Gy. 6 and 20h after BNCT treatment, tumors were resected, fixed and subjected to immunohistochemistry and biochemical analyses. Immunostaining of nuclei showed that double strand break (DSB) marker gamma H2AX staining was high in 20h/+BPA sample but not in 20h/-BPA samples. Poly(ADP-ribose), DSB and single strand break markers of DNA, also demonstrated this tendency. These two markers were observed at low levels in unirradiated tissues or 6h after BNCT either under -BPA and +BPA conditions. HMGB1 level increased in 6h/+BPA but not in 6h/-BPA or 20h/+BPA samples. The persistent staining of γH2AX and poly(ADP-ribose) in +BPA group suggests accumulated DSB damage after BNCT. The early HMGB1 upregulation and γH2AX and poly(ADP-ribose) observed later might be the markers for monitoring the DNA damage induced by BNCT.

Original languageEnglish
Pages (from-to)104-108
Number of pages5
JournalApplied Radiation and Isotopes
Volume88
DOIs
Publication statusPublished - 2014

Fingerprint

Boron Neutron Capture Therapy
DNA Damage
rats
therapy
boron
deoxyribonucleic acid
damage
neutrons
Poly Adenosine Diphosphate Ribose
ribose
adenosine diphosphate
strands
markers
tumors
HMGB1 Protein
Neutrons
staining
Neoplasms
Kazakhstan
Staining and Labeling

Keywords

  • BNCT
  • BPA
  • DNA damage response
  • HMGB1
  • PAR
  • γH2AX

ASJC Scopus subject areas

  • Radiation
  • Medicine(all)

Cite this

Histological and biochemical analysis of DNA damage after BNCT in rat model. / Masutani, Mitsuko; Baiseitov, Diaz; Itoh, Tasuku; Hirai, Takahisa; Berikkhanova, Kulzhan; Murakami, Yasufumi; Zhumadilov, Zhaxybay; Imahori, Yoshio; Hoshi, Masaharu; Itami, Jun.

In: Applied Radiation and Isotopes, Vol. 88, 2014, p. 104-108.

Research output: Contribution to journalArticle

Masutani, M, Baiseitov, D, Itoh, T, Hirai, T, Berikkhanova, K, Murakami, Y, Zhumadilov, Z, Imahori, Y, Hoshi, M & Itami, J 2014, 'Histological and biochemical analysis of DNA damage after BNCT in rat model', Applied Radiation and Isotopes, vol. 88, pp. 104-108. https://doi.org/10.1016/j.apradiso.2014.03.003
Masutani, Mitsuko ; Baiseitov, Diaz ; Itoh, Tasuku ; Hirai, Takahisa ; Berikkhanova, Kulzhan ; Murakami, Yasufumi ; Zhumadilov, Zhaxybay ; Imahori, Yoshio ; Hoshi, Masaharu ; Itami, Jun. / Histological and biochemical analysis of DNA damage after BNCT in rat model. In: Applied Radiation and Isotopes. 2014 ; Vol. 88. pp. 104-108.
@article{8351c585b04044979e7c84eee5a2dfd2,
title = "Histological and biochemical analysis of DNA damage after BNCT in rat model",
abstract = "To understand the mechanism of tumor cell death induced by boron neutron capture therapy (BNCT) and to optimize BNCT condition, we used rat tumor graft models and histological and biochemical analyses were carried out focusing on DNA damage response. Rat lymphosarcoma cells were grafted subcutaneously into male Wister rats. The rats with developed tumors were then treated with neutron beam irradiation 45min after injection of 330mg/kg bodyweight boronophenylalanine (10BPA) (+BPA) or saline control (-BPA). BNCT was carried out in the National Nuclear Center of the Republic of Kazakhstan (neutron flux: 1×109nvt/s, fluence: 6×1011nvt) with the presence of background γ-irradiation of 33Gy. 6 and 20h after BNCT treatment, tumors were resected, fixed and subjected to immunohistochemistry and biochemical analyses. Immunostaining of nuclei showed that double strand break (DSB) marker gamma H2AX staining was high in 20h/+BPA sample but not in 20h/-BPA samples. Poly(ADP-ribose), DSB and single strand break markers of DNA, also demonstrated this tendency. These two markers were observed at low levels in unirradiated tissues or 6h after BNCT either under -BPA and +BPA conditions. HMGB1 level increased in 6h/+BPA but not in 6h/-BPA or 20h/+BPA samples. The persistent staining of γH2AX and poly(ADP-ribose) in +BPA group suggests accumulated DSB damage after BNCT. The early HMGB1 upregulation and γH2AX and poly(ADP-ribose) observed later might be the markers for monitoring the DNA damage induced by BNCT.",
keywords = "BNCT, BPA, DNA damage response, HMGB1, PAR, γH2AX",
author = "Mitsuko Masutani and Diaz Baiseitov and Tasuku Itoh and Takahisa Hirai and Kulzhan Berikkhanova and Yasufumi Murakami and Zhaxybay Zhumadilov and Yoshio Imahori and Masaharu Hoshi and Jun Itami",
year = "2014",
doi = "10.1016/j.apradiso.2014.03.003",
language = "English",
volume = "88",
pages = "104--108",
journal = "Applied Radiation and Isotopes",
issn = "0969-8043",
publisher = "Elsevier",

}

TY - JOUR

T1 - Histological and biochemical analysis of DNA damage after BNCT in rat model

AU - Masutani, Mitsuko

AU - Baiseitov, Diaz

AU - Itoh, Tasuku

AU - Hirai, Takahisa

AU - Berikkhanova, Kulzhan

AU - Murakami, Yasufumi

AU - Zhumadilov, Zhaxybay

AU - Imahori, Yoshio

AU - Hoshi, Masaharu

AU - Itami, Jun

PY - 2014

Y1 - 2014

N2 - To understand the mechanism of tumor cell death induced by boron neutron capture therapy (BNCT) and to optimize BNCT condition, we used rat tumor graft models and histological and biochemical analyses were carried out focusing on DNA damage response. Rat lymphosarcoma cells were grafted subcutaneously into male Wister rats. The rats with developed tumors were then treated with neutron beam irradiation 45min after injection of 330mg/kg bodyweight boronophenylalanine (10BPA) (+BPA) or saline control (-BPA). BNCT was carried out in the National Nuclear Center of the Republic of Kazakhstan (neutron flux: 1×109nvt/s, fluence: 6×1011nvt) with the presence of background γ-irradiation of 33Gy. 6 and 20h after BNCT treatment, tumors were resected, fixed and subjected to immunohistochemistry and biochemical analyses. Immunostaining of nuclei showed that double strand break (DSB) marker gamma H2AX staining was high in 20h/+BPA sample but not in 20h/-BPA samples. Poly(ADP-ribose), DSB and single strand break markers of DNA, also demonstrated this tendency. These two markers were observed at low levels in unirradiated tissues or 6h after BNCT either under -BPA and +BPA conditions. HMGB1 level increased in 6h/+BPA but not in 6h/-BPA or 20h/+BPA samples. The persistent staining of γH2AX and poly(ADP-ribose) in +BPA group suggests accumulated DSB damage after BNCT. The early HMGB1 upregulation and γH2AX and poly(ADP-ribose) observed later might be the markers for monitoring the DNA damage induced by BNCT.

AB - To understand the mechanism of tumor cell death induced by boron neutron capture therapy (BNCT) and to optimize BNCT condition, we used rat tumor graft models and histological and biochemical analyses were carried out focusing on DNA damage response. Rat lymphosarcoma cells were grafted subcutaneously into male Wister rats. The rats with developed tumors were then treated with neutron beam irradiation 45min after injection of 330mg/kg bodyweight boronophenylalanine (10BPA) (+BPA) or saline control (-BPA). BNCT was carried out in the National Nuclear Center of the Republic of Kazakhstan (neutron flux: 1×109nvt/s, fluence: 6×1011nvt) with the presence of background γ-irradiation of 33Gy. 6 and 20h after BNCT treatment, tumors were resected, fixed and subjected to immunohistochemistry and biochemical analyses. Immunostaining of nuclei showed that double strand break (DSB) marker gamma H2AX staining was high in 20h/+BPA sample but not in 20h/-BPA samples. Poly(ADP-ribose), DSB and single strand break markers of DNA, also demonstrated this tendency. These two markers were observed at low levels in unirradiated tissues or 6h after BNCT either under -BPA and +BPA conditions. HMGB1 level increased in 6h/+BPA but not in 6h/-BPA or 20h/+BPA samples. The persistent staining of γH2AX and poly(ADP-ribose) in +BPA group suggests accumulated DSB damage after BNCT. The early HMGB1 upregulation and γH2AX and poly(ADP-ribose) observed later might be the markers for monitoring the DNA damage induced by BNCT.

KW - BNCT

KW - BPA

KW - DNA damage response

KW - HMGB1

KW - PAR

KW - γH2AX

UR - http://www.scopus.com/inward/record.url?scp=84901836830&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84901836830&partnerID=8YFLogxK

U2 - 10.1016/j.apradiso.2014.03.003

DO - 10.1016/j.apradiso.2014.03.003

M3 - Article

VL - 88

SP - 104

EP - 108

JO - Applied Radiation and Isotopes

JF - Applied Radiation and Isotopes

SN - 0969-8043

ER -