TY - JOUR
T1 - Dna strand break properties of protoporphyrin ix by x-ray irradiation against melanoma
AU - Hasegawa, Takema
AU - Takahashi, Junko
AU - Nagasawa, Shinsuke
AU - Doi, Motomichi
AU - Moriyama, Akihiro
AU - Iwahashi, Hitoshi
N1 - Funding Information:
Funding: This work was supported by JSPS KAKENHI Grant Number 18H02705 and 19K22609. This work was also supported by the Canon Foundation Number K16-0073.
Publisher Copyright:
© 2020 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2020/4/1
Y1 - 2020/4/1
N2 - Recent reports have suggested that 5-aminolevulinic acid (5-ALA), which is a precursor to protoporphyrin IX (PpIX), leads to selective accumulation of PpIX in tumor cells and acts as a radiation sensitizer in vitro and in vivo in mouse models of melanoma, glioma, and colon cancer. In this study, we investigated the effect of PpIX under X-ray irradiation through ROS generation and DNA damage. ROS generation by the interaction between PpIX and X-ray was evaluated by two kinds of probes, 3’-(p-aminophenyl) fluorescein (APF) for hydroxyl radical (•OH) detection and dihydroethidium (DHE) for superoxide (O2 •- ). •OH showed an increase, regardless of the dissolved oxygen. Meanwhile, the increase in O2 •- was proportional to the dissolved oxygen. Strand breaks (SBs) of DNA molecule were evaluated by gel electrophoresis, and the enhancement of SBs was observed by PpIX treatment. We also studied the effect of PpIX for DNA damage in cells by X-ray irradiation using a B16 melanoma culture. X-ray irradiation induced γH2AX, DNA double-strand breaks (DSBs) in the context of chromatin, and affected cell survival. Since PpIX can enhance ROS generation even in a hypoxic state and induce DNA damage, combined radiotherapy treatment with 5-ALA is expected to improve therapeutic efficacy for radioresistant tumors.
AB - Recent reports have suggested that 5-aminolevulinic acid (5-ALA), which is a precursor to protoporphyrin IX (PpIX), leads to selective accumulation of PpIX in tumor cells and acts as a radiation sensitizer in vitro and in vivo in mouse models of melanoma, glioma, and colon cancer. In this study, we investigated the effect of PpIX under X-ray irradiation through ROS generation and DNA damage. ROS generation by the interaction between PpIX and X-ray was evaluated by two kinds of probes, 3’-(p-aminophenyl) fluorescein (APF) for hydroxyl radical (•OH) detection and dihydroethidium (DHE) for superoxide (O2 •- ). •OH showed an increase, regardless of the dissolved oxygen. Meanwhile, the increase in O2 •- was proportional to the dissolved oxygen. Strand breaks (SBs) of DNA molecule were evaluated by gel electrophoresis, and the enhancement of SBs was observed by PpIX treatment. We also studied the effect of PpIX for DNA damage in cells by X-ray irradiation using a B16 melanoma culture. X-ray irradiation induced γH2AX, DNA double-strand breaks (DSBs) in the context of chromatin, and affected cell survival. Since PpIX can enhance ROS generation even in a hypoxic state and induce DNA damage, combined radiotherapy treatment with 5-ALA is expected to improve therapeutic efficacy for radioresistant tumors.
KW - DNA double-strand break
KW - Melanoma
KW - Photodynamic therapy
KW - Protoporphyrin IX
KW - Radiodynamic therapy
KW - Radiotherapy
KW - Reactive oxygen species (ROS)
KW - X-ray
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U2 - 10.3390/ijms21072302
DO - 10.3390/ijms21072302
M3 - Article
C2 - 32225109
AN - SCOPUS:85082791239
SN - 1661-6596
VL - 21
JO - International journal of molecular sciences
JF - International journal of molecular sciences
IS - 7
M1 - 2302
ER -