A single high-dose irradiation changes accumulation of methotrexate and gene expression levels of SLC and ABC transporters in cancer cells

Kakeru Sato; Tatsuya Seki; Asuka Mizutani; Yuka Muranaka; Shiho Hirota; Kodai Nishi; Kana Yamazaki; Ryuichi Nishii; Takeo Nakanishi; Ikumi Tamai; Keiichi Kawai; Masato Kobayashi

doi:10.3389/fphar.2022.1069321

A single high-dose irradiation changes accumulation of methotrexate and gene expression levels of SLC and ABC transporters in cancer cells

Front Pharmacol. 2023 Jan 11:13:1069321. doi: 10.3389/fphar.2022.1069321. eCollection 2022.

Authors

Affiliations

¹ Division of Health Sciences, Graduate School of Medical Sciences, Kanazawa University, Kanazawa, Japan.
² Faculty of Health Sciences, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kanazawa, Japan.
³ Department of Molecular Imaging and Theranostics, Institute for Quantum Medical Science, Quantum Life and Medical Science Directorate, National Institutes for Quantum Science and Technology, Chiba, Japan.
⁴ Faculty of Pharmacy, Takasaki University of Health and Welfare, Takasaki, Gunma, Japan.
⁵ Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical and Health Sciences, Kanazawa University, Kanazawa, Japan.
⁶ Biomedical Imaging Research Center, University of Fukui, Fukui, Japan.

Abstract

Chemoradiotherapy is frequently used to treat cancer. Stereotactic body radiotherapy (SBRT) is a single high-dose radiotherapy used to treat a variety of cancers. The anticancer drug methotrexate (MTX) shows affinity for solute carrier (SLC) and ATP-binding cassette (ABC) transporters. This study investigated relationships between accumulation of methotrexate and gene expression levels of solute carrier and ATP-binding cassette transporters in cancer cells after a single and high-dose X-ray irradiation. Cancer cell lines were selected from lung and cervical cancer cell line that are commonly used for stereotactic body radiotherapy and effective with methotrexate. We examined expression levels of organic anion-transporting polypeptide (OATP)1B1, OATP1B3, OATP1B7, and organic anion transporter (OAT)1 as solute carrier transporters and multidrug resistance-associated protein (MRP)1 and MRP2 as ATP-binding cassette transporters, using real-time polymerase chain reaction and accumulation of ³H-MTX in cancer cells after 10-Gy irradiation, assuming stereotactic body radiotherapy. Cells were divided into three groups: Control without irradiation; 4 h after irradiation; and 24 h after irradiation. In control, gene expression levels of OAT1 in all cells was below the limit of measurement. After irradiation, gene expression levels of OATP1B1/1B3/1B7 showed changes in each cell line. Gene expression levels of MRP1/2 tended to increase after irradiation. Gene expression levels of OATP1B1/1B3/1B7 were much lower than those of MRP1/2. Accumulation of ³H-MTX tended to decrease over time after irradiation. Irradiation of cancer cells thus alters gene expression levels of both solute carrier transporters (OATP1B1/1B3/1B7) and ABC transporters (MRP1/2) and decreases accumulation of ³H-MTX in cancer cells over time due to elevated expression of MRP1/2.

Keywords: ABC transporters; SLC transporter; chemoradiotherapy; irradiation; methotrexate; stereotactic body radiotherapy; x-ray.

Grants and funding

This study was funded in part by Grants-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (Nos 21H02865, 22H03016, and 22K19504) and the Network-type Joint Usage/Research Center for Radiation Disaster Medical Science.