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Antitumor effects of cytoplasmic delivery of an innate adjuvant receptor ligand, poly(I:C), on human breast cancer

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Abstract

Innate adjuvant receptors are expressed in immune cells and some types of cancers. If antitumor therapies targeting these receptors are established, it is likely that they will be therapeutically beneficial because antitumor effects and immune-cell activation can be induced simultaneously. In this study, we tested this possibility of using an innate adjuvant receptor ligand, polyinosinic–polycytidylic acid [poly(I:C)], to treat human breast cancer cell lines. Three breast cancer cell lines (MCF-7, MDA-MB-231, and BT-549) were used in this study. Poly(I:C) was transfected into these cancer cells to stimulate melanoma differentiation–associated gene (MDA) 5, which is a cytoplasmic adjuvant receptor. Poly(I:C) transfection significantly reduced the viability of all cell lines in a manner partially dependent on MDA5. Flow cytometeric analyses and immunoblot assays revealed that the antitumor effect depended on both caspase-dependent apoptosis and c-Myc- and cyclinD1-dependent growth arrest. Interestingly, poly(I:C) transfection was accompanied by autophagy, which is thought to protect cancer cells from apoptosis after poly(I:C) transfection. In a xenograft mouse model, local transfection of poly(I:C) significantly inhibited the growth of xenografted MDA-MB-231 cells. Our findings indicate that cytoplasmic delivery of poly(I:C) can induce apoptosis and growth arrest of human breast cancer cells, and that therapy-associated autophagy prevents apoptosis. The results of this study suggest that the innate adjuvant receptors are promising targets and that their ligands could serve as antitumor reagents, which have the potential to simultaneously induce antitumor effects and activate immune cells.

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Abbreviations

CFSE:

Carboxyfluorescein diacetate, succinimidyl ester

DC:

Dendritic cell

IFN:

Interferon

MDA:

Melanoma differentiation–associated gene

PI:

Propidium iodide

poly(I:C):

Polyinosinic–polycytidylic acid

siRNA:

Small interfering RNA

TLR:

Toll-like receptor

WST-8:

2-(2-Methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2, 4-disulfophenyl)-2H-tetrazolium monosodium salt

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Acknowledgments

We thank Ms. Yasuko Moriyama for her technical assistance. This study was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology, Japan (no. 18591449 to M.H. and no. 23701074 to N.H.).

Conflict of interest

None.

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Authors and Affiliations

  1. Department of Immunology, Shimane University Faculty of Medicine, Izumo, Shimane, 693-8501, Japan

    Touko Inao, Nanae Harashima, Hiroyuki Monma & Mamoru Harada

  2. Department of Surgery, Shimane University Faculty of Medicine, Shimane, Japan

    Touko Inao, Hiroyuki Monma, Masayuki Itakura, Tsuneo Tanaka & Yoshitsugu Tajima

  3. Division of Pathophysiological and Experimental Pathology, Department of Pathology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan

    Shinji Okano

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  1. Touko Inao
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Correspondence to Mamoru Harada.

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Inao, T., Harashima, N., Monma, H. et al. Antitumor effects of cytoplasmic delivery of an innate adjuvant receptor ligand, poly(I:C), on human breast cancer. Breast Cancer Res Treat 134, 89–100 (2012). https://doi.org/10.1007/s10549-011-1930-3

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