Chemerin mediated neutrophil infiltration in oral squamous cell carcinoma and predicted poor prognosis

doi:10.19439/j.sjos.2023.02.009

Abstract

Abstract: PURPOSE: To explore the effect of Chemerin in oral squamous cell carcinoma (OSCC) tissue on neutrophils infiltration and its possible molecular mechanism. METHODS: The relationship between Chemerin expression and neutrophils density was assessed via double immunohistochemistry staining.The chemotactic effect of Chemerin on neutrophils in OSCC was detected by transwell assay, real-time quantitative PCR(qRT-PCR), Western blot, enzyme-linked immunosorbent assay(ELISA) and flow cytometry. The data were statistically analyzed using SPSS 23.0 software package. The relationship between Chemerin expression and neutrophils density was assessed using Spearman rank correlation analysis. ChemR23 knockout efficiency and chemotactic index were calculated by ANOVA. The relationship between Chemerin expression, neutrophils density and clinicopathological factors was analyzed by Mann-Whitney test. Kaplan-Meier test and Log rank test were used for survival analysis, and risk factors affecting the survival of OSCC patients was assessed using Cox regression model. RESULTS: Double immunohistochemistry staining showed that overexpression of Chemerin was significantly correlated with increased neutrophils infiltration in OSCC(P=0.023), and strong Chemerin expression and high neutrophils density were associated with higher clinical stage(P<0.001), cervical lymph node metastasis (P<0.001) and tumor recurrence (P=0.002). Kaplan-Meier survival analysis showed that patients in the strong Chemerin expression + high neutrophils density group had shortened cancer-related overall survival time and disease-free survival time compared with the other two groups. Transwell assay results showed that both OSCC cells and R-Chemerin had a significant chemotactic effect on dHL-60 cells; knockdown of ChemR23 suppressed Chemerin-induced chemotaxis to dHL-60 cells. CONCLUSIONS: Overexpression of Chemerin in OSCC tissue chemoattracts more neutrophils to tumor sites through its receptor ChemR23 and is related to poor clinical prognosis.

Key words: Chemerin, Neutrophils, Oral squamous cell carcinoma, ChemR23

CLC Number:

R739.8

HU Xiao-yuan, WANG Ning, ZHANG Xiao-ye, YANG Qian, GAO Fei, WANG Cheng-qin, SHANG Wei, XIANG Feng-gang, FENG Yuan-yong. Chemerin mediated neutrophil infiltration in oral squamous cell carcinoma and predicted poor prognosis[J]. Shanghai Journal of Stomatology, 2023, 32(2): 158-165.

References

[1] Siegel R, Naishadham D, Jemal A.Cancer statistics for Hispanics/Latinos, 2012[J]. CA Cancer J Clin, 2012, 62(5): 283-298.
[2] Sano D, Myers JN.Metastasis of squamous cell carcinoma of the oral tongue[J]. Cancer Metastasis Rev, 2007, 26(3-4): 645-662.
[3] Coffelt SB, Wellenstein MD, de Visser KE. Neutrophils in cancer: neutral no more[J]. Nat Rev Cancer, 2016, 16(7): 431-446.
[4] Huang H, Zhang H, Onuma AE, et al.Neutrophil elastase and neutrophil extracellular traps in the tumor microenvironment[J]. Adv Exp Med Biol, 2020, 1263: 13-23.
[5] Ferland DJ, Watts SW.Chemerin: a comprehensive review elucidating the need for cardiovascular research[J]. Pharmacol Res, 2015, 99: 351-361.
[6] Treeck O, Buechler C, Ortmann O.Chemerin and cancer[J]. Int J Mol Sci, 2019, 20(15): 3750-3765.
[7] Weigert J, Obermeier F, Neumeier M, et al.Circulating levels of chemerin and adiponectin are higher in ulcerative colitis and chemerin is elevated in Crohn's disease[J]. Inflamm Bowel Dis, 2010, 16(4): 630-637.
[8] Sotiropoulos GP, Dalamaga M, Antonakos G, et al.Chemerin as a biomarker at the intersection of inflammation, chemotaxis, coagulation, fibrinolysis and metabolism in resectable non-small cell lung cancer[J]. Lung Cancer, 2018, 125: 291-299.
[9] Zabel BA, Allen SJ, Kulig P, et al.Chemerin activation by serine proteases of the coagulation, fibrinolytic, and inflammatory cascades[J]. J Biol Chem, 2005, 280(41): 34661-34666.
[10] Mariani F, Roncucci L.Chemerin/chemR23 axis in inflammation onset and resolution[J]. Inflamm Res, 2015, 64(2): 85-95.
[11] Wang N, Wang QJ, Feng YY, et al.Overexpression of chemerin was associated with tumor angiogenesis and poor clinical outcome in squamous cell carcinoma of the oral tongue[J]. Clin Oral Investig, 2014, 18(3): 997-1004.
[12] Wang N, Feng Y, Wang Q, et al.Neutrophils infiltration in the tongue squamous cell carcinoma and its correlation with CEACAM1 expression on tumor cells[J]. PLoS One, 2014, 9(2): e89991.
[13] Zhang W, Gu J, Chen J, et al.Interaction with neutrophils promotes gastric cancer cell migration and invasion by inducing epithelial-mesenchymal transition[J]. Oncol Rep, 2017, 38(5): 2959-2966.
[14] de Souza LF, Pearson AG, Pace PE, et al. Peroxiredoxin expression and redox status in neutrophils and HL-60 cells[J]. Free Radic Biol Med, 2019, 135: 227-234.
[15] Wang N, Wang Q, Chi J, et al.Carcinoembryonic antigen cell adhesion molecule 1 inhibits the antitumor effect of neutrophils in tongue squamous cell carcinoma[J]. Cancer Sci, 2019, 110(2): 519-529.
[16] Wculek SK, Malanchi I.Neutrophils support lung colonization of metastasis-initiating breast cancer cells[J]. Nature, 2015, 528(7582): 413-417.
[17] Masucci MT, Minopoli M, Carriero MV. Tumor associated neutrophils. Their Role in tumorigenesis, metastasis, prognosis and therapy [J]. Front Oncol, 2019, 9: 1146.e1-1146.e16.
[18] Mollaoglu G, Jones A, Wait SJ, et al. The Lineage-defining transcription factors SOX2 and NKX2-1 determine lung cancer cell fate and shape the tumor immune microenvironment [J]. Immunity, 2018, 49(4): 764-779.e9.
[19] Sell H, Laurencikiene J, Taube A, et al.Chemerin is a novel adipocyte-derived factor inducing insulin resistance in primary human skeletal muscle cells[J]. Diabetes, 2009, 58(12): 2731-2740.
[20] Kaur J, Adya R, Tan BK, et al.Identification of chemerin receptor (ChemR23) in human endothelial cells: chemerin-induced endothelial angiogenesis[J]. Biochem Biophys Res Commun, 2010, 391(4): 1762-1768.
[21] Vermi W, Riboldi E, Wittamer V, et al.Role of ChemR23 in directing the migration of myeloid and plasmacytoid dendritic cells to lymphoid organs and inflamed skin[J]. J Exp Med, 2005, 201(4): 509-515.
[22] Coffelt SB, Kersten K, Doornebal CW, et al.IL-17-producing γδ T cells and neutrophils conspire to promote breast cancer metastasis[J]. Nature, 2015, 522(7556): 345-348.
[23] Gordon-Weeks AN, Lim SY, Yuzhalin AE, et al.Neutrophils promote hepatic metastasis growth through fibroblast growth factor 2-dependent angiogenesis in mice[J]. Hepatology, 2017, 65(6): 1920-1935.
[24] Lazennec G, Richmond A.Chemokines and chemokine receptors: new insights into cancer-related inflammation[J]. Trends Mol Med, 2010, 16(3): 133-144.
[25] Mantovani A, Allavena P, Sica A, et al.Cancer-related inflammation[J]. Nature, 2008, 454(7203): 436-444.
[26] Del Prete A, Sozio F, Schioppa T, et al.The atypical receptor CCRL2 is essential for lung cancer immune surveillance[J]. Cancer Immunol Res, 2019, 7(11): 1775-1788.