Abstract: Numerous studies have demonstrated that the high expression of CXC motif chemokine ligand 16 (CXCL16) in cancer correlates with poor prognosis, as well as tumor cell proliferation, migration, and invasion. While CXCL16 can serve as a tumor biomarker, the underlying mechanism in modulating head and neck squamous cell carcinoma (HNSCC) remains unclear. In this study, the aimed was to investigate the CXCL16 expression in HNSCC and to uncover the potential underlying mechanism. Hereby, we determined the high expression of CXCL16 in The Cancer Genome Atlas (TCGA) database, as well as in tissue samples from patients with HNSCC at our central hospital and from HNSCC cell lines. The results showed that CXCL16 knockdown inhibited the proliferation, migration, and invasion of HNSCC cells. Mechanistically, transcriptome sequencing revealed that CXCL16 may affect HNSCC cell growth by regulating the antioxidant pathway of glutathione peroxidase 1 (GPX1). The reactive oxygen species (ROS) levels were elevated in small interfering CXCL16 (si-CXCL16) cells, which may contribute to the inhibition of cell proliferation, migration, and invasion. Moreover, treatment of cells with the GPX1 inhibitor eldecalcitol (ED-71) revealed that HNSCC cell growth was significantly inhibited in the synergistic group of si-CXCL16 and GPX1 inhibitor compared to the si-CXCL16 group. In conclusion, CXCL16 contributed to the development of HNSCC cells by modulating the GPX1-mediated antioxidant pathway. Thus, targeting cellular CXCL16 expression seems to be a promising strategy for treating HNSCC.

CXCL16通过调节GPX1介导的抗氧化水平促进头颈部鳞状细胞癌增殖

[1]AbelS, HundhausenC, MentleinR, et al., 2004. The transmembrane CXC-chemokine ligand 16 is induced by IFN-γ and TNF-α and shed by the activity of the disintegrin-like metalloproteinase ADAM10. J Immunol, 172(10):6362-6372.

[2]BeckerTM, JuvikJA, 2016. The role of glucosinolate hydrolysis products from brassica vegetable consumption in inducing antioxidant activity and reducing cancer incidence. Diseases, 4(2):22.

[3]BrayF, FerlayJ, SoerjomataramI, et al., 2018. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin, 68(6):394-424.

[4]BrocksteinB, HarafDJ, RademakerAW, et al., 2004. Patterns of failure, prognostic factors and survival in locoregionally advanced head and neck cancer treated with concomitant chemoradiotherapy: a 9-year, 337-patient, multi-institutional experience. Ann Oncol, 15(8):1179-1186.

[5]Chalabi-DcharM, Cassant-SourdyS, DulucC, et al., 2015. Loss of somatostatin receptor subtype 2 promotes growth of KRAS-induced pancreatic tumors in mice by activating PI3K signaling and overexpression of CXCL16. Gastroenterology, 148(7):1452-1465.

[6]ChoSW, KimYA, SunHJ, et al., 2016. CXCL16 signaling mediated macrophage effects on tumor invasion of papillary thyroid carcinoma. Endocr Relat Cancer, 23(2):113-124.

[7]ColladoA, MarquesP, EscuderoP, et al., 2018. Functional role of endothelial CXCL16/CXCR6-platelet-leucocyte axis in angiotensin II-associated metabolic disorders. Cardiovasc Res, 114(13):1764-1775.

[8]DequanterD, DokR, KoolenL, et al., 2017. Prognostic significance of glutathione peroxidase levels (GPx1) in head and neck cancers. Front Oncol, 7:84.

[9]FanS, TangQL, LinYJ, et al., 2011. A review of clinical and histological parameters associated with contralateral neck metastases in oral squamous cell carcinoma. Int J Oral Sci, 3(4):180-191.

[10]GaoZR, LingXY, ShiCY, et al., 2022. Tumor immune checkpoints and their associated inhibitors. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(10):823-843.

[11]GriffithJW, SokolCL, LusterAD, 2014. Chemokines and chemo

[12]kine receptors: positioning cells for host defense and immunity. Annu Rev Immunol, 32:659-702.

[13]GuanJZ, XuX, QiuG, et al., 2023. Cellular hierarchy framework based on single-cell/multi-patient sample sequencing reveals metabolic biomarker PYGL as a therapeutic target for HNSCC. J Exp Clin Cancer Res, 42:162.

[14]HanJ, FuRJ, ChenC, et al., 2021. CXCL16 promotes gastric cancer tumorigenesis via ADAM10-dependent CXCL16/CXCR6 axis and activates Akt and MAPK signaling pathways. Int J Biol Sci, 17(11):2841-2852.

[15]HattermannK, GebhardtH, KrossaS, et al., 2016. Transmembrane chemokines act as receptors in a novel mechanism termed inverse signaling. eLife, 5:e10820.

[16]HsiehCY, LinCC, HuangYW, et al., 2022. Macrophage secretory IL-1β promotes docetaxel resistance in head and neck squamous carcinoma via SOD2/CAT-ICAM1 signaling. JCI Insight, 7(23):e157285.

[17]HuWD, LiuY, ZhouWH, et al., 2014. CXCL16 and CXCR6 are coexpressed in human lung cancer in vivo and mediate the invasion of lung cancer cell lines in vitro. PLoS ONE, 9(6):e99056.

[18]HuangC, DingGY, GuCY, et al., 2012. Decreased selenium-binding protein 1 enhances glutathione peroxidase 1 activity and downregulates HIF-1α to promote hepatocellular carcinoma invasiveness. Clin Cancer Res, 18(11):3042-3053.

[19]IzquierdoMC, Martin-ClearyC, Fernandez-FernandezB, et al., 2014. CXCL16 in kidney and cardiovascular injury. Cytokine Growth Factor Rev, 25(3):317-325.

[20]JiangYY, GuoHY, TongT, et al., 2022. lncRNA lnc-POP1-1 upregulated by VN1R5 promotes cisplatin resistance in head and neck squamous cell carcinoma through interaction with MCM5. Mol Ther, 30(1):448-467.

[21]KarakiS, BlancC, TranT, et al., 2021. CXCR6 deficiency impairs cancer vaccine efficacy and CD8⁺ resident memory T-cell recruitment in head and neck and lung tumors. J Immunother Cancer, 9(3):e001948.

[22]KassabRB, ElhenawyAA, AbdulrahmanTheyab, et al., 2023. Modulation of inflammatory, oxidative, and apoptotic stresses mediates the renoprotective effect of daidzein against glycerol-induced acute kidney injury in rats. Environ Sci Pollut Res Int, 30(56):119016-119033.

[23]KhanP, FatimaM, KhanMA, et al., 2022. Emerging role of chemokines in small cell lung cancer: road signs for metastasis, heterogeneity, and immune response. Semin Cancer Biol, 87:117-126.

[24]KimMJ, SunHJ, SongYS, et al., 2019. CXCL16 positively correlated with M2-macrophage infiltration, enhanced angiogenesis, and poor prognosis in thyroid cancer. Sci Rep, 9:13288.

[25]KorbeckiJ, Bajdak-RusinekK, KupnickaP, et al., 2021. The role of CXCL16 in the pathogenesis of cancer and other diseases. Int J Mol Sci, 22(7):3490.

[26]LeiFJ, ChiangJY, ChangHJ, et al., 2023. Cellular and exosomal GPx1 are essential for controlling hydrogen peroxide balance and alleviating oxidative stress in hypoxic glioblastoma. Redox Biol, 65:102831.

[27]LeporeF, D'AlessandroG, AntonangeliF, et al., 2018. CXCL16/CXCR6 axis drives microglia/macrophages phenotype in physiological conditions and plays a crucial role in glioma. Front Immunol, 9:2750.

[28]LiuH, YangZJ, LuWP, et al., 2020. Chemokines and chemokine receptors: a new strategy for breast cancer therapy. Cancer Med, 9(11):3786-3799.

[29]LiuHY, WangGQ, WuT, et al., 2022. Efficacy and safety of eldecalcitol for osteoporosis: a meta-analysis of randomized controlled trials. Front Endocrinol (Lausanne), 13:854439.

[30]LivakKJ, SchmittgenTD, 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods, 25(4):402-408.

[31]LuYP, KouYY, GaoY, et al., 2023. Eldecalcitol inhibits the progression of oral cancer by suppressing the expression of GPx-1. Oral Dis, 29(2):615-627.

[32]MaYS, XuX, LuoM, 2017. CXCR6 promotes tumor cell proliferation and metastasis in osteosarcoma through the Akt pathway. Cell Immunol, 311:80-85.

[33]MatloubianM, DavidA, EngelS, et al., 2000. A transmembrane CXC chemokine is a ligand for HIV-coreceptor Bonzo. Nat Immunol, 1(4):298-304.

[34]MatsushitaK, ToiyamaY, TanakaK, et al., 2012. Soluble CXCL16 in preoperative serum is a novel prognostic marker and predicts recurrence of liver metastases in colorectal cancer patients. Ann Surg Oncol, 19(Suppl 3):518-527.

[35]MeiXY, QiDS, ZhangT, et al., 2020. Inhibiting MARSs reduces hyperhomocysteinemia-associated neural tube and congenital heart defects. EMBO Mol Med, 12(3):e9469.

[36]MeiZ, HuangJW, QiaoB, et al., 2020. Immune checkpoint pathways in immunotherapy for head and neck squamous cell carcinoma. Int J Oral Sci, 12:16.

[37]MoloneyJN, CotterTG, 2018. ROS signalling in the biology of cancer. Semin Cell Dev Biol, 80:50-64.

[38]MoreiraD, SampathS, WonH, et al., 2021. Myeloid cell-targeted STAT3 inhibition sensitizes head and neck cancers to radiotherapy and T cell-mediated immunity. J Clin Invest, 131(2):e137001.

[39]PeiJ, PanXY, WeiGH, et al., 2023. Research progress of glutathione peroxidase family (GPX) in redoxidation. Front Pharmacol, 14:1147414.

[40]RichardsenE, NessN, Melbø-JørgensenC, et al., 2015. The prognostic significance of CXCL16 and its receptor C-X-C chemokine receptor 6 in prostate cancer. Am J Pathol, 185(10):2722-2730.

[41]RuffinAT, LiH, VujanovicL, et al., 2023. Improving head and neck cancer therapies by immunomodulation of the tumour microenvironment. Nat Rev Cancer, 23(3):173-188.

[42]Saddawi-KonefkaR, O'FarrellA, FarajiF, et al., 2022. Lymphatic-preserving treatment sequencing with immune checkpoint inhibition unleashes cDC1-dependent antitumor immunity in HNSCC. Nat Commun, 13:4298.

[43]SiegelRL, MillerKD, JemalA, 2020. Cancer statistics, 2020. CA Cancer J Clin, 70(1):7-30.

[44]SiuLL, EvenC, MesíaR, et al., 2019. Safety and efficacy of durvalumab with or without tremelimumab in patients with PD-L1-low/negative recurrent or metastatic HNSCC: the phase 2 CONDOR randomized clinical trial. JAMA Oncol, 5(2):195-203.

[45]TangYC, HsiaoJR, JiangSS, et al., 2021. c-MYC-directed NRF2 drives malignant progression of head and neck cancer via glucose-6-phosphate dehydrogenase and transketolase activation. Theranostics, 11(11):5232-5247.

[46]TsaiCF, ChenGW, ChenYC, et al., 2022. Regulatory effects of quercetin on M1/M2 macrophage polarization and oxidative/antioxidative balance. Nutrients, 14(1):67.

[47]WuW, GengZX, BaiHR, et al., 2021. Ammonium ferric citrate induced ferroptosis in non-small-cell lung carcinoma through the inhibition of GPX4-GSS/GSR-GGT axis activity. Int J Med Sci, 18(8):1899-1909.

[48]ZhangGS, WangQ, QiXL, et al., 2022. OShnscc: a novel user-friendly online survival analysis tool for head and neck squamous cell carcinoma based on RNA expression profiles and long-term survival information. J Zhejiang Univ-Sci B (Biomed & Biotechnol), 23(3):249-257.

[49]ZhaoYJ, WangH, ZhouJD, et al., 2022. Glutathione peroxidase GPX1 and its dichotomous roles in cancer. Cancers (Basel), 14(10):2560.