The TLR7 agonist imiquimod induces anti-cancer effects via autophagic cell death and enhances anti-tumoral and systemic immunity during radiotherapy for melanoma

// Jeong Hyun Cho 1, * , Hyo-Ji Lee 1, * , Hyun-Jeong Ko 2 , Byung-Il Yoon 3 , Jongseon Choe 4 , Keun-Cheol Kim 1 , Tae-Wook Hahn 3 , Jeong A. Han 5 , Sun Shim Choi 6 , Young Mee Jung 7 , Kee-Ho Lee 8 , Yun-Sil Lee 9 , Yu-Jin Jung 1 1 Department of Biological Sciences, Kangwon National University, Chuncheon, Republic of Korea 2 College of Pharmacy, Kangwon National University, Chuncheon, Republic of Korea 3 Department of Veterinary Medicine, Kangwon National University, Chuncheon, Republic of Korea 4 Department of Microbiology, School of Medicine, Kangwon National University, Chuncheon, Republic of Korea 5 Department of Biochemistry and Molecular Biology, School of Medicine, Kangwon National University, Chuncheon, Republic of Korea 6 Department of Medical Biotechnology, College of Biomedical Science, Kangwon National University, Chuncheon, Republic of Korea 7 Department of Chemistry, Kangwon National University, Chuncheon, Republic of Korea 8 Division of Radiation Cancer Research, Korea Institute of Radiological and Medical Sciences, Nowon-gu, Seoul, Republic of Korea 9 Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seodaemun-gu, Seoul, Republic of Korea * These authors have contributed equally to this work Correspondence to: Yu-Jin Jung, email: yjjung@kangwon.ac.kr Keywords: melanoma, TLR7, imiquimod (IMQ), autophagy, radiotherapy Received: February 17, 2016 Accepted: January 23, 2017 Published: February 15, 2017 ABSTRACT Toll-like receptor (TLR) ligands are strongly considered immune-adjuvants for cancer immunotherapy and have been shown to exert direct anti-cancer effects. This study was performed to evaluate the synergistic anti-cancer and anti-metastatic effects of the TLR7 agonist imiquimod (IMQ) during radiotherapy for melanoma. The pretreatment of B16F10 or B16F1 cells with IMQ combined with γ-ionizing radiation (IR) led to enhanced cell death via autophagy, as demonstrated by increased expression levels of autophagy-related genes, and an increased number of autophagosomes in both cell lines. The results also confirmed that the autophagy process was accelerated via the reactive oxygen species (ROS)-mediated MAPK and NF-κB signaling pathway in the cells pretreated with IMQ combined with IR. Mice subcutaneously injected with melanoma cells showed a reduced tumor growth rate after treatment with IMQ and IR. Treatment with 3-methyladenine (3-MA), ameliorated the anti-cancer effect of IMQ combined with IR. Additionally, the combination therapy enhanced anti-cancer immunity, as demonstrated by an increased number of CD8 + T cells and decreased numbers of regulatory T cells (Treg) and myeloid-derived suppressor cells (MDSCs) in the tumor lesions. Moreover, the combination therapy decreased the number of metastatic nodules in the lungs of mice that were injected with B16F10 cells via the tail vein. In addition, the combination therapy enhanced systemic anti-cancer immunity by increasing the abundances of T cell populations expressing IFN-γ and TNF-α. Therefore, these findings suggest that IMQ could serve as a radiosensitizer and immune booster during radiotherapy for melanoma patients.