Toll‑like receptor 4 plays a tumor‑suppressive role in cutaneous squamous cell carcinoma

Mikami,Erina; Kudo,Mitsuhiro; Ohashi,Ryuji; Kawahara,Kiyoko; Kawamoto,Yoko; Teduka,Kiyoshi; Fujii,Takenori; Kitamura,Taeko; Kure,Shoko; Ishino,Kousuke; Sakatani,Takashi; Wada,Ryuichi; Saeki,Hidehisa; Naito,Zenya

doi:10.3892/ijo.2019.4790

Toll‑like receptor 4 plays a tumor‑suppressive role in cutaneous squamous cell carcinoma

Authors:
- Erina Mikami
- Mitsuhiro Kudo
- Ryuji Ohashi
- Kiyoko Kawahara
- Yoko Kawamoto
- Kiyoshi Teduka
- Takenori Fujii
- Taeko Kitamura
- Shoko Kure
- Kousuke Ishino
- Takashi Sakatani
- Ryuichi Wada
- Hidehisa Saeki
- Zenya Naito
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Affiliations: Department of Integrated Diagnostic Pathology, Nippon Medical School, Tokyo 113‑8603, Japan, Department of Diagnostic Pathology, Nippon Medical School Musashikosugi Hospital, Kawasaki, Kanagawa 211‑8533, Japan, Department of Dermatology, Nippon Medical School, Tokyo 113‑8603, Japan
Published online on: April 17, 2019 https://doi.org/10.3892/ijo.2019.4790
Pages: 2179-2188

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Abstract

Toll‑like receptor 4 (TLR4), a key regulator of the innate immune system, is expressed not only in immune cells, but also in a number of cancer cells. A biological role for TLR4 in cutaneous squamous cell carcinoma (SCC), however, is unclear. In this study, we first examined TLR4 expression and localization in cases of SCC, actinic keratosis (AK) and Bowen's disease (BD) by immunohistochemistry. TLR4 expression was significantly higher in the SCC than in the AK or BD tissues. We then determined the TLR4 expression level in vivo, in 3 histological subtypes of SCC. TLR4 expression in poorly differentiated SCC was significantly lower compared with that of the moderately and well‑differentiated type. In addition, the CD44 immunoreactivity tended to be high in the cell membrane of poorly differentiated SCC. Of note, poorly differentiated SCC is a risk factor of unfavorable outcomes in affected patients. We then assessed the biological role of TLR4 in HSC‑1 and HSC‑5 SCC cells and HaCaT human keratinocytes. TLR4 knockdown by transfection with siRNA accelerated HSC‑1 and HaCaT cell migration and invasion compared to the control siRNA‑transfected cells. TLR4 knockdown resulted in an increased CD44 expression and in an enhanced filopodia protrusion formation, particularly in HSC‑1. On the whole, these results suggest that a reduced TLR4 expression enhances the malignant features in SCC cases and cultured SCC cell lines. TLR4 may thus play an anti‑tumor role in cutaneous SCC.

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1	Burton KA, Ashack KA and Khachemoune A: Cutaneous Squamous Cell Carcinoma: A Review of High-Risk and Metastatic Disease. Am J Clin Dermatol. 17:491–508. 2016. View Article : Google Scholar : PubMed/NCBI
2	Karia PS, Han J and Schmults CD: Cutaneous squamous cell carcinoma: Estimated incidence of disease, nodal metastasis, and deaths from disease in the United States, 2012. J Am Acad Dermatol. 68:957–966. 2013. View Article : Google Scholar : PubMed/NCBI
3	Zhou M, McFarland-Mancini MM, Funk HM, Husseinzadeh N, Mounajjed T and Drew AF: Toll-like receptor expression in normal ovary and ovarian tumors. Cancer Immunol Immunother. 58:1375–1385. 2009. View Article : Google Scholar : PubMed/NCBI
4	Sun Y, Wu C, Ma J, Yang Y, Man X, Wu H and Li S: Toll-like receptor 4 promotes angiogenesis in pancreatic cancer via PI3K/AKT signaling. Exp Cell Res. 347:274–282. 2016. View Article : Google Scholar : PubMed/NCBI
5	Dong YQ, Lu CW, Zhang L, Yang J, Hameed W and Chen W: Toll-like receptor 4 signaling promotes invasion of hepatocellular carcinoma cells through MKK4/JNK pathway. Mol Immunol. 68:671–683. 2015. View Article : Google Scholar : PubMed/NCBI
6	Ye K, Wu Y, Sun Y, Lin J and Xu J: TLR4 siRNA inhibits proliferation and invasion in colorectal cancer cells by downregulating ACAT1 expression. Life Sci. 155:133–139. 2016. View Article : Google Scholar : PubMed/NCBI
7	Zou Y, Qin F, Chen J, Meng J, Wei L, Wu C, Zhang Q, Wei D, Chen X, Wu H, et al: sTLR4/MD-2 complex inhibits colorectal cancer in vitro and in vivo by targeting LPS. Oncotarget. 7:52032–52044. 2016. View Article : Google Scholar : PubMed/NCBI
8	Eiró N, Ovies C, Fernandez-Garcia B, Álvarez-Cuesta CC, González L, González LO and Vizoso FJ: Expression of TLR3, 4, 7 and 9 in cutaneous malignant melanoma: Relationship with clinicopathological characteristics and prognosis. Arch Dermatol Res. 305:59–67. 2013. View Article : Google Scholar
9	Janda J, Burkett NB, Blohm-Mangone K, Huang V, Curiel-Lewandrowski C, Alberts DS, Petricoin EF III, Calvert VS, Einspahr J, Dong Z, et al: Resatorvid-based Pharmacological Antagonism of Cutaneous TLR4 Blocks UV-induced NF-κB and AP-1 Signaling in Keratinocytes and Mouse Skin. Photochem Photobiol. 92:816–825. 2016. View Article : Google Scholar : PubMed/NCBI
10	Blohm-Mangone K, Burkett NB, Tahsin S, Myrdal PB, Aodah A, Ho B, Janda J, McComas M, Saboda K, Roe DJ, et al: Pharmacological TLR4 Antagonism Using Topical Resatorvid Blocks Solar UV-Induced Skin Tumorigenesis in SKH-1 Mice. Cancer Prev Res (Phila). 11:265–278. 2018. View Article : Google Scholar
11	Iotzova-Weiss G, Freiberger SN, Johansen P, Kamarachev J, Guenova E, Dziunycz PJ, Roux GA, Neu J and Hofbauer GFL: TLR4 as a negative regulator of keratinocyte proliferation. PLoS One. 12:e01856682017. View Article : Google Scholar : PubMed/NCBI
12	Szatmary Z: Molecular biology of toll-like receptors. Gen Physiol Biophys. 31:357–366. 2012. View Article : Google Scholar : PubMed/NCBI
13	Mai CW, Kang YB and Pichika MR: Should a Toll-like receptor 4 (TLR-4) agonist or antagonist be designed to treat cancer? TLR-4: Its expression and effects in the ten most common cancers. Onco Targets Ther. 6:1573–1587. 2013.PubMed/NCBI
14	Dajon M, Iribarren K and Cremer I: Toll-like receptor stimulation in cancer: A pro- and anti-tumor double-edged sword. Immunobiology. 222:89–100. 2017. View Article : Google Scholar
15	Miller LS: Toll-like receptors in skin. Adv Dermatol. 24:71–87. 2008. View Article : Google Scholar : PubMed/NCBI
16	Weng H, Deng Y, Xie Y, Liu H and Gong F: Expression and significance of HMGB1, TLR4 and NF-κB p65 in human epidermal tumors. BMC Cancer. 13:3112013. View Article : Google Scholar
17	Burns EM and Yusuf N: Toll-like receptors and skin cancer. Front Immunol. 5:1352014. View Article : Google Scholar : PubMed/NCBI
18	Yusuf N, Nasti TH, Long JA, Naseemuddin M, Lucas AP, Xu H and Elmets CA: Protective role of Toll-like receptor 4 during the initiation stage of cutaneous chemical carcinogenesis. Cancer Res. 68:615–622. 2008. View Article : Google Scholar : PubMed/NCBI
19	Karvinen S, Kosma V-MM, Tammi MII and Tammi R: Hyaluronan, CD44 and versican in epidermal keratinocyte tumours. Br J Dermatol. 148:86–94. 2003. View Article : Google Scholar : PubMed/NCBI
20	Hartmann-Petersen S, Tammi RH, Tammi MI and Kosma VM: Depletion of cell surface CD44 in nonmelanoma skin tumours is associated with increased expression of matrix metallopro-teinase 7. Br J Dermatol. 160:1251–1257. 2009. View Article : Google Scholar : PubMed/NCBI
21	Erfani E, Roudi R, Rakhshan A, Sabet MN, Shariftabrizi A and Madjd Z: Comparative expression analysis of putative cancer stem cell markers CD44 and ALDH1A1 in various skin cancer subtypes. Int J Biol Markers. 31:e53–e61. 2016. View Article : Google Scholar
22	Senbanjo LT and Chellaiah MA: CD44: A Multifunctional Cell Surface Adhesion Receptor Is a Regulator of Progression and Metastasis of Cancer Cells. Front Cell Dev Biol. 5:182017. View Article : Google Scholar : PubMed/NCBI
23	Bourguignon LYW: Matrix hyaluronan-activated CD44 signaling promotes keratinocyte activities and improves abnormal epidermal functions. Am J Pathol. 184:1912–1919. 2014. View Article : Google Scholar : PubMed/NCBI
24	Bourguignon LYW, Wong G, Earle CA and Xia W: Interaction of low molecular weight hyaluronan with CD44 and toll-like receptors promotes the actin filament-associated protein 110-actin binding and MyD88-NFκB signaling leading to proinflammatory cytokine/chemokine production and breast tumor invasion. Cytoskeleton (Hoboken). 68:671–693. 2011. View Article : Google Scholar
25	Elder DE, Massi D, Scolyer R and Willemze R: WHO Classification of Skin Tumours Fourth Edition. WHO Classification of Tumours. 11. IARC; Lyon: 2018
26	Kondo S and Aso K: Establishment of a cell line of human skin squamous cell carcinoma in vitro. Br J Dermatol. 105:125–132. 1981. View Article : Google Scholar : PubMed/NCBI
27	Hozumi Y, Kondo S, Shimoura T and Aso K: Human squamous cell carcinoma from skin: Establishment and characterization of a new cell line (HSC-5). J Dermatol. 17:143–148. 1990. View Article : Google Scholar : PubMed/NCBI
28	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. Methods. 25:402–408. 2001. View Article : Google Scholar
29	Yanofsky VR, Mercer SE and Phelps RG: Histopathological variants of cutaneous squamous cell carcinoma: A review. J Skin Cancer. 2011:2108132011. View Article : Google Scholar : PubMed/NCBI
30	Motaparthi K, Kapil JP and Velazquez EF: Cutaneous Squamous Cell Carcinoma: Review of the Eighth Edition of the American Joint Committee on Cancer Staging Guidelines, Prognostic Factors, and Histopathologic Variants. Adv Anat Pathol. 24:171–194. 2017. View Article : Google Scholar : PubMed/NCBI
31	Jacquemet G, Hamidi H and Ivaska J: Filopodia in cell adhesion, 3D migration and cancer cell invasion. Curr Opin Cell Biol. 36:23–31. 2015. View Article : Google Scholar : PubMed/NCBI
32	Dang S, Peng Y, Ye L, Wang Y, Qian Z, Chen Y, Wang X, Lin Y, Zhang X, Sun X, et al: Stimulation of TLR4 by LMW-HA induces metastasis in human papillary thyroid carcinoma through CXCR7. Clin Dev Immunol. 2013:7125612013. View Article : Google Scholar : PubMed/NCBI
33	Yu L, Wang L and Chen S: Dual character of Toll-like receptor signaling: Pro-tumorigenic effects and anti-tumor functions. Biochim Biophys Acta. 1835:144–154. 2013.
34	Haricharan S and Brown P: TLR4 has a TP53-dependent dual role in regulating breast cancer cell growth. Proc Natl Acad Sci USA. 112:E3216–E3225. 2015. View Article : Google Scholar : PubMed/NCBI
35	Chen C, Zhao S, Karnad A and Freeman JW: The biology and role of CD44 in cancer progression: Therapeutic implications. J Hematol Oncol. 11:642018. View Article : Google Scholar : PubMed/NCBI
36	Brierley JD, Gospodarowicz MK and Wittekind C: TNM Classification of Malignant Tumours. 8th edition. Union for International Cancer Control (UICC); Geneva: 2016