Overexpression of B7‑H4 promotes renal cell carcinoma progression by recruiting tumor‑associated neutrophils via upregulation of CXCL8

Li,Anqi; Zhang,Ningyue; Zhao,Zhiming; Chen,Yali; Zhang,Liang

doi:10.3892/ol.2020.11701

Overexpression of B7‑H4 promotes renal cell carcinoma progression by recruiting tumor‑associated neutrophils via upregulation of CXCL8

Authors:
- Anqi Li
- Ningyue Zhang
- Zhiming Zhao
- Yali Chen
- Liang Zhang
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Affiliations: Department of Biological Pharmacy, College of Pharmaceutical Sciences, Soochow University, Suzhou, Jiangsu 215123, P.R. China
Published online on: June 5, 2020 https://doi.org/10.3892/ol.2020.11701
Pages: 1535-1544
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The immune checkpoint molecule B7 family member H4 (B7‑H4) plays a similar role to programmed death‑ligand 1 in tumor immune evasion by regulating T‑cell‑mediated immune responses. However, besides the role in T‑cell immunity, B7‑H4 also affects tumor cell biology by promoting tumor cell proliferation, metastasis and angiogenesis. In order to explore the effect of B7‑H4 on tumor cell biology, it is necessary to investigate the gene expression profile when B7‑H4 is overexpressed. In the present study, 786‑O cells were transfected to stably express B7‑H4. A microarray technique was subsequently used to screen B7‑H4‑related differentially expressed genes (DEGs) in B7‑H4/786‑O cells compared with negative control (NC)/786‑O cells. The protein expression of the upregulated DEGs, including non‑metastatic cells 5, NME/NM23 family member 5 (NME5), membrane metalloendopeptidase (MME), vascular non‑inflammatory molecule 1 (VNN1), matrix metalloproteinase (MMP) 7, tumor necrosis factor, C‑X‑C motif chemokine ligand (CXCL) 8, CXCL1 and C‑C motif chemokine ligand (CCL) 2, was investigated using western blotting. Kidney renal papillary cell carcinoma mRNA‑sequencing data obtained from The Cancer Genome Atlas revealed that chemokines, including CXCL1/2/3, CXCL8, MMP7 and CCL20, were positively correlated with B7‑H4 gene expression. Furthermore, 59 clinical renal cell carcinoma tissues were collected and analyzed by immunohistochemical staining. The results revealed the positive correlation of B7‑H4 with CCL20 and CXCL8, and validated the DEGs identified in tumor cell lines. 786‑O transfectants were inoculated into non‑obese diabetic/severe combined immunodeficiency mice, and tumor growth was investigated. B7‑H4 overexpression promoted tumor growth and administration of anti‑CXCL8 antibody reversed this effect. Furthermore, B7‑H4 overexpression increased the number of tumor‑infiltrating neutrophils while inhibition of CXCL8 abrogated this effect. These data indicated that recruitment of neutrophils in the tumor microenvironment by CXCL8 serves an important role in the tumor promotion effect of B7‑H4. The present study revealed a novel mechanism of B7‑H4 in tumor promotion in addition to T cell inhibition.

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1	Ferlay J, Soerjomataram I, Dikshit R, Eser S, Mathers C, Rebelo M, Parkin DM, Forman D and Bray F: Cancer incidence and mortality worldwide: Sources, methods and major patterns in GLOBOCAN 2012. Int J Cancer. 136:E359–E386. 2015. View Article : Google Scholar : PubMed/NCBI
2	Rooney MS, Shukla SA, Wu CJ, Getz G and Hacohen N: Molecular and genetic properties of tumors associated with local immune cytolytic activity. Cell. 160:48–61. 2015. View Article : Google Scholar : PubMed/NCBI
3	Santoni M, Berardi R, Amantini C, Burattini L, Santini D, Santoni G and Cascinu S: Role of natural and adaptive immunity in renal cell carcinoma response to VEGFR-TKIs and mTOR inhibitor. Int J Cancer. 134:2772–2777. 2014. View Article : Google Scholar : PubMed/NCBI
4	Motzer RJ, Hutson TE, Tomczak P, Michaelson MD, Bukowski RM, Rixe O, Oudard S, Negrier S, Szczylik C, Kim ST, et al: Sunitinib versus interferon alfa in metastatic renal-cell carcinoma. N Engl J Med. 356:115–124. 2007. View Article : Google Scholar : PubMed/NCBI
5	Motzer RJ, Escudier B, McDermott DF, George S, Hammers HJ, Srinivas S, Tykodi SS, Sosman JA, Procopio G, Plimack ER, et al: Nivolumab versus everolimus in advanced renal-cell carcinoma. N Engl J Med. 373:1803–1813. 2015. View Article : Google Scholar : PubMed/NCBI
6	Gill DM, Hahn AW, Hale P and Maughan BL: Overview of current and future first-line systemic therapy for metastatic clear cell renal cell carcinoma. Curr Treat Options Oncol. 19:62018. View Article : Google Scholar : PubMed/NCBI
7	Kawashima A, Uemura M and Nonomura N: Importance of multiparametric evaluation of immune-related T-cell markers in renal-cell carcinoma. Clin Genitourin Cancer. 17:e1147–e1152. 2019. View Article : Google Scholar : PubMed/NCBI
8	Sica GL, Choi IH, Zhu G, Tamada K, Wang SD, Tamura H, Chapoval AI, Flies DB, Bajorath J and Chen L: B7-H4, a molecule of the B7 family, negatively regulates T cell immunity. Immunity. 18:849–861. 2003. View Article : Google Scholar : PubMed/NCBI
9	Zang X, Loke P, Kim J, Murphy K, Waitz R and Allison JP: B7×: A widely expressed B7 family member that inhibits T cell activation. Proc Natl Acad Sci USA. 100:10388–10392. 2003. View Article : Google Scholar : PubMed/NCBI
10	Tang Z, Li C, Kang B, Gao G, Li C and Zhang Z: GEPIA: A web server for cancer and normal gene expression profiling and interactive analyses. Nucleic Acids Res. 45:W98–W102. 2017. View Article : Google Scholar : PubMed/NCBI
11	Li C, Zhan Y, Ma X, Fang H and Gai X: B7-H4 facilitates proliferation and metastasis of colorectal carcinoma cell through PI3K/Akt/mTOR signaling pathway. Clin Exp Med. Oct 29–2019.doi: 10.1007/s10238-019-00590-7 (Epub ahead of print).
12	Genova C, Boccardo S, Mora M, Rijavec E, Biello F, Rossi G, Tagliamento M, Dal Bello MG, Coco S, Alama A, et al: Correlation between B7-H4 and survival of non-small-cell lung cancer patients treated with nivolumab. J Clin Med. 8(pii): E15662019. View Article : Google Scholar
13	Gruosso T, Gigoux M, Manem VSK, Bertos N, Zuo D, Perlitch I, Saleh SMI, Zhao H, Souleimanova M, Johnson RM, et al: Spatially distinct tumor immune microenvironments stratify triple-negative breast cancers. J Clin Invest. 129:1785–1800. 2019. View Article : Google Scholar : PubMed/NCBI
14	Krambeck AE, Thompson RH, Dong H, Lohse CM, Park ES, Kuntz SM, Leibovich BC, Blute ML, Cheville JC, Kwon ED, et al: B7-H4 expression in renal cell carcinoma and tumor vasculature: Associations with cancer progression and survival. Proc Natl Acad Sci USA. 103:10391–10396. 2006. View Article : Google Scholar : PubMed/NCBI
15	Xie N, Cai JB, Zhang L, Zhang PF, Shen YH, Yang X, Lu JC, Gao DM, Kang Q, Liu LX, et al: Upregulation of B7-H4 promotes tumor progression of intrahepatic cholangiocarcinoma. Cell Death Dis. 8:32052017. View Article : Google Scholar : PubMed/NCBI
16	Zhu J, Chu BF, Yang YP, Zhang SL, Zhuang M, Lu WJ and Liu YB: B7-H4 expression is associated with cancer progression and predicts patient survival in human thyroid cancer. Asian Pac J Cancer Prev. 14:3011–3015. 2013. View Article : Google Scholar : PubMed/NCBI
17	Prasad DV, Richards S, Mai XM and Dong C: B7S1, a novel B7 family member that negatively regulates T cell activation. Immunity. 18:863–873. 2003. View Article : Google Scholar : PubMed/NCBI
18	Li J, Lee Y, Li Y, Jiang Y, Lu H, Zang W, Zhao X, Liu L, Chen Y, Tan H, et al: Co-inhibitory molecule B7 superfamily member 1 expressed by tumor-infiltrating myeloid cells induces dysfunction of Anti-tumor CD8(+) T cells. Immunity. 48:773–786.e5. 2018. View Article : Google Scholar : PubMed/NCBI
19	Chen C, Qu QX, Xie F, Zhu WD, Zhu YH and Huang JA: Analysis of B7-H4 expression in metastatic pleural adenocarcinoma and therapeutic potential of its antagonists. BMC Cancer. 17:6522017. View Article : Google Scholar : PubMed/NCBI
20	Chen C, Zhu WD, Xie F and Huang JA: Nuclear localization of B7-H4 in pulmonary adenocarcinomas presenting as a solitary pulmonary nodule. Oncotarget. 7:58563–58568. 2016. View Article : Google Scholar : PubMed/NCBI
21	Zhang L, Wu H, Lu D, Li G, Sun C, Song H, Li J, Zhai T, Huang L, Hou C, et al: The costimulatory molecule B7-H4 promote tumor progression and cell proliferation through translocating into nucleus. Oncogene. 32:5347–5358. 2013. View Article : Google Scholar : PubMed/NCBI
22	Jeon YK, Park SG, Choi IW, Lee SW, Lee SM and Choi I: Cancer cell-associated cytoplasmic B7-H4 is induced by hypoxia through hypoxia-inducible factor-1a and promotes cancer cell proliferation. Biochem Biophys Res Commun. 459:277–283. 2015. View Article : Google Scholar : PubMed/NCBI
23	Xia F, Zhang Y, Xie L, Jiang H, Zeng H, Chen C, Liu L, He X, Hao X, Fang X, et al: B7-H4 enhances the differentiation of murine leukemia-initiating cells via the PTEN/AKT/RCOR2/RUNX1 pathways. Leukemia. 31:2260–2264. 2017. View Article : Google Scholar : PubMed/NCBI
24	Tang Z, Li C, Kang B, Gao G and Zhang Z: GEPIA: A web server for cancer and normal gene expression profiling and interactive analyses. Nucleic Acids Res. 45:W98–W102. 2017. View Article : Google Scholar : PubMed/NCBI
25	Allred DC, Clark GM, Elledge R, Fuqua SA, Brown RW, Chamness GC, Osborne CK and McGuire WL: Association of p53 protein expression with tumor cell proliferation rate and clinical outcome in node-negative breast cancer. J Natl Cancer Inst. 85:200–206. 1993. View Article : Google Scholar : PubMed/NCBI
26	Wang DH, Lee HS, Yoon D, Berry G, Wheeler TM, Sugarbaker DJ, Kheradmand F, Engleman E and Burt BM: Progression of EGFR-Mutant lung adenocarcinoma is driven by alveolar macrophages. Clin Cancer Res. 23:778–788. 2017. View Article : Google Scholar : PubMed/NCBI
27	Ogawa R, Yamamoto T, Hirai H, Hanada K, Kiyasu Y, Nishikawa G, Mizuno R, Inamoto S, Itatani Y, Sakai Y and Kawada K: Loss of SMAD4 promotes colorectal cancer progression by recruiting tumor-associated neutrophils via CXCL1/8-CXCR2 axis. Clin Cancer Res. 25:2287–2899. 2019. View Article : Google Scholar
28	Wang L, Heng X, Lu Y, Cai Z, Yi Q and Che F: Could B7-H4 serve as a target to activate anti-cancer immunity? Int Immunopharmacol. 38:97–103. 2016. View Article : Google Scholar : PubMed/NCBI
29	Bregar A, Deshpande A, Grange C, Zi T, Stall J, Hirsch H, Reeves J, Sathyanarayanan S, Growdon WB and Rueda BR: Characterization of immune regulatory molecules B7-H4 and PD-L1 in low and high grade endometrial tumors. Gynecol Oncol. 145:446–452. 2017. View Article : Google Scholar : PubMed/NCBI
30	Han S, Wang Y, Shi X, Zong L, Liu L, Zhang J, Qian Q, Jin J, Ma Y, Cui B, et al: Negative roles of B7-H3 and B7-H4 in the microenvironment of cervical cancer. Exp Cell Res. 371:222–230. 2018. View Article : Google Scholar : PubMed/NCBI
31	Huang H, Li C and Ren G: Clinical significance of the B7-H4 as a novel prognostic marker in breast cancer. Gene. 623:24–28. 2017. View Article : Google Scholar : PubMed/NCBI
32	MacGregor HL and Ohashi PS: Molecular pathways: Evaluating the potential for B7-H4 as an immunoregulatory target. Clin Cancer Res. 23:2934–2941. 2017. View Article : Google Scholar : PubMed/NCBI
33	Tan Z and Shen W: Prognostic role of B7-H4 in patients with non-small cell lung cancer: A meta-analysis. Oncotarget. 8:27137–27144. 2017. View Article : Google Scholar : PubMed/NCBI
34	Wang L, Yang C, Liu XB, Wang L and Kang FB: B7-H4 overexpression contributes to poor prognosis and drug-resistance in triple-negative breast cancer. Cancer Cell Int. 18:1002018. View Article : Google Scholar : PubMed/NCBI
35	Qian Y, Sang Y, Wang FX, Hong B, Wang Q, Zhou X, Weng T, Wu Z, Zheng M, Zhang H and Yao H: Prognostic significance of B7-H4 expression in matched primary pancreatic cancer and liver metastases. Oncotarget. 7:72242–72249. 2016. View Article : Google Scholar : PubMed/NCBI
36	Han S, Li Y, Zhang J, Liu L, Chen Q, Qian Q, Li S and Zhang Y: Roles of immune inhibitory molecule B7-H4 in cervical cancer. Oncology Rep. 37:2308–2316. 2017. View Article : Google Scholar
37	Chen X, Wang W, Man H, Li P and Shan B: Increased B7-H4 expression during esophageal squamous cell carcinogenesis is associated with IL-6/STAT3 signaling pathway activation in mice. Oncol Lett. 13:2207–2215. 2017. View Article : Google Scholar : PubMed/NCBI
38	Qian Y, Feng L, Wu W, Weng T, Hu C, Hong B, Wang FXC, Shen L, Wang Q, Jin X and Yao H: MicroRNA expression profiling of pancreatic cancer cell line L3.6p1 following B7-H4 Knockdown. Cell Physiol Biochem. 44:494–504. 2017. View Article : Google Scholar : PubMed/NCBI
39	Azuma T, Sato Y, Ohno T, Azuma M and Kume H: Serum soluble B7-H4 is a prognostic marker for patients with non-metastatic clear cell renal cell carcinoma. PLoS One. 13:e01997192018. View Article : Google Scholar : PubMed/NCBI
40	Jacquelot N, Duong CPM, Belz GT and Zitvogel L: Targeting chemokines and chemokine receptors in melanoma and other cancers. Front Immunol. 9:24802018. View Article : Google Scholar : PubMed/NCBI
41	Zhang Y, Roth TL, Gray EE, Chen H, Rodda LB, Liang Y, Ventura P, Villeda S, Crocker PR and Cyster JG: Migratory and adhesive cues controlling innate-like lymphocyte surveillance of the pathogen-exposed surface of the lymph node. Elife. 5(pii): e181562016. View Article : Google Scholar : PubMed/NCBI
42	Ramirez-Valle F, Gray EE and Cyster JG: Inflammation induces dermal Vγ4+ γδT17 memory-like cells that travel to distant skin and accelerate secondary IL-17-driven responses. Proc Natl Acad Sci USA. 112:8046–8051. 2015. View Article : Google Scholar : PubMed/NCBI
43	Hartwig T, Pantelyushin S, Croxford AL, Kulig P and Becher B: Dermal IL-17-producing γδ T cells establish long-lived memory in the skin. Eur J Immunol. 45:3022–3033. 2015. View Article : Google Scholar : PubMed/NCBI
44	Vilgelm AE and Richmond A: Chemokines modulate immune surveillance in tumorigenesis, metastasis, and response to immunotherapy. Front Immunol. 10:3332019. View Article : Google Scholar : PubMed/NCBI
45	Susek KH, Karvouni M, Alici E and Lundqvist A: The role of CXC chemokine receptors 1–4 on immune cells in the tumor microenvironment. Front Immunol. 9:21592018. View Article : Google Scholar : PubMed/NCBI
46	Young D, Das N, Anowai A and Dufour A: Matrix metalloproteases as influencers of the Cells' social media. Int J Mol Sci. 20(pii): E38472019. View Article : Google Scholar : PubMed/NCBI
47	Liao CH, Chang WS, Hu PS, Wu HC, Hsu SW, Liu YF, Liu SP, Hung HS, Bau DT and Tsai CW: The Contribution of MMP-7 promoter polymorphisms in renal cell carcinoma. In Vivo. 31:631–635. 2017. View Article : Google Scholar : PubMed/NCBI
48	Zhang Z, Dong T, Fu Y, Zhou W, Tian X, Chen G and Liu S: MMP-11 promotes papillary thyroid cell proliferation and invasion via the NF-κB pathway. J Cell Biochem. Sep 1–2018.doi: 10.1002/jcb.27500 (Epub ahead of print).