Neutrophil extracellular traps promote gastric cancer metastasis by inducing epithelial‑mesenchymal transition

Zhu,Tong; Zou,Xiaoming; Yang,Chunfa; Li,Liangliang; Wang,Bing; Li,Rong; Li,Hongxuan; Xu,Zhangxuan; Huang,Di; Wu,Qingyun

doi:10.3892/ijmm.2021.4960

Neutrophil extracellular traps promote gastric cancer metastasis by inducing epithelial‑mesenchymal transition

Authors:
- Tong Zhu
- Xiaoming Zou
- Chunfa Yang
- Liangliang Li
- Bing Wang
- Rong Li
- Hongxuan Li
- Zhangxuan Xu
- Di Huang
- Qingyun Wu
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Affiliations: The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China, Shuangyashan Shuangkuang Hospital, Shuangyashan, Heilongjiang 155100, P.R. China, Department of Pharmacology, Harbin Medical University, Harbin, Heilongjiang 150086, P.R. China
Published online on: May 13, 2021 https://doi.org/10.3892/ijmm.2021.4960
Article Number: 127
Copyright: © Zhu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The risks of tumor recurrence following the successful resection of the primary tumor have been known for decades; however, the precise mechanisms underlying treatment failures remain unknown. The formation of neutrophil extracellular traps (NETs) has increasingly been demonstrated to be associated with thrombi formation in cancer patients, as well as with the development and metastasis of cancer. The present study demonstrated that the level of peripheral blood NETs in patients with gastric cancer (GC) was associated with tumor progression, and patients with stage III/IV disease exhibited significant differences compared with the healthy controls and patients with stage I/II disease, which may be associated with an increased risk of metastasis. In addition, plasma from patients with stage III/IV GC was more prone to stimulate neutrophils to form NETs; thus, it was hypothesized that the formation of NETs may be affected by the tumor microenvironment. A higher deposition of NETs in GC tissues compared with normal resection margins was also identified. In vitro, following treatment with phorbol myristate acetate, which promotes the formation of NETs, or with DNAse‑1/GSK‑484, which inhibits the formation of NETs, it was found that the tumor migratory ability was altered; however, no significant changes were observed in cell proliferation and cell cycle progression. Epithelial‑mesenchymal transition (EMT) is a key event associated with dissemination and metastasis in GC pathogenesis. Finally, the present study demonstrated that NETs promote a more aggressive mesenchymal phenotype and promote the progression of GC in vitro and in vivo. On the whole, to the best of our knowledge, the present study reports a previously unknown role of NETs in the regulation of GC, which is associated with EMT and migration. Therefore, targeting NETs may prove to be therapeutically beneficial.

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1	Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA and Jemal A: Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 68:394–424. 2018. View Article : Google Scholar : PubMed/NCBI
2	Chen W, Zheng R, Baade PD, Zhang S, Zeng H, Bray F, Jemal A, Yu XQ and He J: Cancer statistics in China, 2015. CA Cancer J Clin. 66:115–132. 2016. View Article : Google Scholar : PubMed/NCBI
3	Arneth B: Tumor microenvironment. Medicina (Kaunas). 56:152019. View Article : Google Scholar
4	Quail DF and Joyce JA: Microenvironmental regulation of tumor progression and metastasis. Nat Med. 19:1423–1437. 2013. View Article : Google Scholar : PubMed/NCBI
5	Gonzalez H, Hagerling C and Werb Z: Roles of the immune system in cancer: From tumor initiation to metastatic progression. Genes Dev. 32:1267–1284. 2018. View Article : Google Scholar : PubMed/NCBI
6	Zhang X, Shi H, Yuan X, Jiang P, Qian H and Xu W: Tumor-derived exosomes induce N2 polarization of neutrophils to promote gastric cancer cell migration. Mol Cancer. 17:1462018. View Article : Google Scholar : PubMed/NCBI
7	Shaul ME and Fridlender ZG: Tumour-associated neutrophils in patients with cancer. Nat Rev Clin Oncol. 16:601–620. 2019. View Article : Google Scholar : PubMed/NCBI
8	Coffelt SB, Wellenstein MD and de Visser KE: Neutrophils in cancer: Neutral no more. Nat Rev Cancer. 16:4314462016. View Article : Google Scholar
9	Liu Y and Liu L: The pro-tumor effect and the anti-tumor effect of neutrophils extracellular traps. Biosci Trends. 13:469–475. 2020. View Article : Google Scholar
10	Demers M, Krause DS, Schatzberg D, Martinod K, Voorhees JR, Fuchs TA, Scadden DT and Wagner DD: Cancers predispose neutrophils to release extracellular DNA traps that contribute to cancer-associated thrombosis. Proc Natl Acad Sci USA. 109:13076–13081. 2012. View Article : Google Scholar : PubMed/NCBI
11	Cedervall J, Hamidi A and Olsson AK: Platelets, NETs and cancer. Thromb Res. 164(Suppl 1): S148–S152. 2018. View Article : Google Scholar : PubMed/NCBI
12	Berger-Achituv S, Brinkmann V, Abed UA, Kühn LI, Ben-Ezra J, Elhasid R and Zychlinsky A: A proposed role for neutrophil extracellular traps in cancer immunoediting. Front Immunol. 4:482013. View Article : Google Scholar : PubMed/NCBI
13	Park J, Wysocki RW, Amoozgar Z, Maiorino L, Fein MR, Jorns J, Schott AF, Kinugasa-Katayama Y, Lee Y, Won NH, et al: Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps. Sci Transl Med. 8:361ra1382016. View Article : Google Scholar : PubMed/NCBI
14	Najmeh S, Cools-Lartigue J, Rayes RF, Gowing S, Vourtzoumis P, Bourdeau F, Giannias B, Berube J, Rousseau S, Ferri LE and Spicer JD: Neutrophil extracellular traps sequester circulating tumor cells via β1-integrin mediated interactions. Int J Cancer. 140:2321–2330. 2017. View Article : Google Scholar : PubMed/NCBI
15	Cools-Lartigue J, Spicer J, McDonald B, Gowing S, Chow S, Giannias B, Bourdeau F, Kubes P and Ferri L: Neutrophil extracellular traps sequester circulating tumor cells and promote metastasis. J Clin Invest. 123:3446–3458. 2013. View Article : Google Scholar :
16	Grilz E, Mauracher LM, Posch F, Königsbrügge O, Zöchbauer-Müller S, Marosi C, Lang I, Pabinger I and Ay C: Citrullinated histone H3, a biomarker for neutrophil extracellular trap formation, predicts the risk of mortality in patients with cancer. Br J Haematol. 186:311–320. 2019. View Article : Google Scholar : PubMed/NCBI
17	Erpenbeck L and Schön MP: Neutrophil extracellular traps: Protagonists of cancer progression? Oncogene. 36:2483–2490. 2017. View Article : Google Scholar
18	Claushuis TA, de Stoppelaar SF, Stroo I, Roelofs JJ, Ottenhoff R, van der Poll T and Van't Veer C: Thrombin contributes to protective immunity in pneumonia-derived sepsis via fibrin polymerization and platelet-neutrophil interactions. J Thromb Haemost. 15:744–757. 2017. View Article : Google Scholar : PubMed/NCBI
19	Arelaki S, Arampatzioglou A, Kambas K, Papagoras C, Miltiades P, Angelidou I, Mitsios A, Kotsianidis I, Skendros P, Sivridis E, et al: Gradient infiltration of neutrophil extracellular traps in colon cancer and evidence for their involvement in tumour growth. PLoS One. 11:e01544842016. View Article : Google Scholar : PubMed/NCBI
20	Schedel F, Mayer-Hain S, Pappelbaum KI, Metze D, Stock M, Goerge T, Loser K, Sunderkötter C, Luger TA and Weishaupt C: Evidence and impact of neutrophil extracellular traps in malignant melanoma. Pigment Cell Melanoma Res. 33:63–73. 2020. View Article : Google Scholar
21	Gupta GP and Massagué J: Cancer metastasis: Building a framework. Cell. 127:679–695. 2006. View Article : Google Scholar : PubMed/NCBI
22	Pastushenko I and Blanpain C: EMT transition states during tumor progression and metastasis. Trends Cell Biol. 29:212–226. 2019. View Article : Google Scholar
23	Zhang W, Gu J, Chen J, Zhang P, Ji R, Qian H, Xu W and Zhang X: Interaction with neutrophils promotes gastric cancer cell migration and invasion by inducing epithelial-mesenchymal transition. Oncol Rep. 38:2959–2966. 2017. View Article : Google Scholar : PubMed/NCBI
24	Li S, Cong X, Gao H, Lan X, Li Z, Wang W, Song S, Wang Y, Li C, Zhang H, et al: Correction to: Tumor-associated neutrophils induce EMT by IL-17a to promote migration and invasion in gastric cancer cells. J Exp Clin Cancer Res. 38:1772019. View Article : Google Scholar : PubMed/NCBI
25	Rayes RF, Vourtzoumis P, Bou Rjeily M, Seth R, Bourdeau F, Giannias B, Berube J, Huang YH, Rousseau S, Camilleri-Broet S, et al: Neutrophil extracellular trap-associated CEACAM1 as a putative therapeutic target to prevent metastatic progression of colon carcinoma. J Immunol. 204:2285–2294. 2020. View Article : Google Scholar : PubMed/NCBI
26	American Joint Committee on Cancer (AJCC): Cancer Staging Manual. AJCC; Chicago, IL: 2016, https://cancerstaging.org/references-tools/deskreferences/pages/default.aspx.
27	Albrengues J, Shields MA, Ng D, Park CG, Ambrico A, Poindexter ME, Upadhyay P, Uyeminami DL, Pommier A, Küttner V, et al: Neutrophil extracellular traps produced during inflammation awaken dormant cancer cells in mice. Science. 361:eaao42272018. View Article : Google Scholar : PubMed/NCBI
28	Li B, Liu Y, Hu T, Zhang Y, Zhang C, Li T, Wang C, Dong Z, Novakovic VA, Hu T and Shi J: Neutrophil extracellular traps enhance procoagulant activity in patients with oral squamous cell carcinoma. J Cancer Res Clin Oncol. 145:1695–1707. 2019. View Article : Google Scholar : PubMed/NCBI
29	Hu Y, Li H, Yan R, Wang C, Wang Y, Zhang C, Liu M, Zhou T, Zhu W, Zhang H, et al: Increased neutrophil activation and plasma DNA levels in patients with pre-eclampsia. Thromb Haemost. 118:2064–2073. 2018. View Article : Google Scholar : PubMed/NCBI
30	Schimmel M, Nur E, Biemond BJ, van Mierlo GJ, Solati S, Brandjes DP, Otten HM, Schnog JJ, Zeerleder S and Curama Study Group: Nucleosomes and neutrophil activation in sickle cell disease painful crisis. Haematologica. 98:1797–1803. 2013. View Article : Google Scholar : PubMed/NCBI
31	Yoo DG, Floyd M, Winn M, Moskowitz SM and Rada B: NET formation induced by Pseudomonas aeruginosa cystic fibrosis isolates measured as release of myeloperoxidase-DNA and neutrophil elastase-DNA complexes. Immunol Lett. 160:186–194. 2014. View Article : Google Scholar : PubMed/NCBI
32	Stakos DA, Kambas K, Konstantinidis T, Mitroulis I, Apostolidou E, Arelaki S, Tsironidou V, Giatromanolaki A, Skendros P, Konstantinides S and Ritis K: Expression of functional tissue factor by neutrophil extracellular traps in culprit artery of acute myocardial infarction. Eur Heart J. 36:1405–1414. 2015. View Article : Google Scholar : PubMed/NCBI
33	Abed AU and Brinkmann V: Immunofluorescence labelling of human and murine neutrophil extracellular traps in paraffin-embedded tissue. J Vis Exp. 151:e601152019.
34	Owen GR, Häkkinen L, Wu C and Larjava H: A reproducible technique for specific labeling of antigens using preformed fluorescent molecular IgG-F(ab′)2 complexes from primary anti-bodies of the same species. Microsc Res Tech. 73:623–630. 2010.
35	Hu Z, Murakami T, Tamura H, Reich J, Kuwahara-Arai K, Iba T, Tabe Y and Nagaoka I: Neutrophil extracellular traps induce IL-1β production by macrophages in combination with lipopolysaccharide. Int J Mol Med. 39:549–558. 2017. View Article : Google Scholar : PubMed/NCBI
36	Brinkmann V, Reichard U, Goosmann C, Fauler B, Uhlemann Y, Weiss DS, Weinrauch Y and Zychlinsky A: Neutrophil extracellular traps kill bacteria. Science. 303:1532–1535. 2004. View Article : Google Scholar : PubMed/NCBI
37	Nauseef WM and Borregaard N: Neutrophils at work. Nat Immunol. 15:602–611. 2014. View Article : Google Scholar : PubMed/NCBI
38	Yang C, Sun W, Cui W, Li X, Yao J, Jia X, Li C, Wu H, Hu Z and Zou X: Procoagulant role of neutrophil extracellular traps in patients with gastric cancer. Int J Clin Exp Pathol. 8:14075–14086. 2015.
39	Marin Oyarzún CP, Carestia A, Lev PR, Glembotsky AC, Castro Ríos MA, Moiraghi B, Molinas FC, Marta RF, Schattner M and Heller PG: Neutrophil extracellular trap formation and circulating nucleosomes in patients with chronic myeloproliferative neoplasms. Sci Rep. 6:387382016. View Article : Google Scholar : PubMed/NCBI
40	Oklu R, Sheth RA, Wong KHK, Jahromi AH and Albadawi H: Neutrophil extracellular traps are increased in cancer patients but does not associate with venous thrombosis. Cardiovasc Diagn Ther. 7(Suppl 3): S140–S149. 2017. View Article : Google Scholar
41	Guglietta S and Rescigno M: Hypercoagulation and complement: Connected players in tumor development and metastases. Semin Immunol. 28:578–586. 2016. View Article : Google Scholar : PubMed/NCBI
42	Brinkmann V: Neutrophil extracellular traps in the second decade. J Innate Immun. 10:414–421. 2018. View Article : Google Scholar : PubMed/NCBI
43	Wu L, Liang Y, Zhang C, Wang X, Ding X, Huang C and Liang H: Prognostic significance of lymphovascular infiltration in overall survival of gastric cancer patients after surgery with curative intent. Chin J Cancer Res. 31:785–796. 2019. View Article : Google Scholar : PubMed/NCBI
44	Abdol Razak N, Elaskalani O and Metharom P: Pancreatic cancer-induced neutrophil extracellular traps: A potential contributor to cancer-associated thrombosis. Int J Mol Sci. 18:4872017. View Article : Google Scholar :
45	Richardson JJR, Hendrickse C, Gao-Smith F and Thickett DR: Neutrophil extracellular trap production in patients with colorectal cancer in vitro. Int J Inflam. 2017:49150622017. View Article : Google Scholar : PubMed/NCBI
46	Zhang Y, Wang C, Yu M, Zhao X, Du J, Li Y, Jing H, Dong Z, Kou J, Bi Y, et al: Neutrophil extracellular traps induced by activated platelets contribute to procoagulant activity in patients with colorectal cancer. Thromb Res. 180:87–97. 2019. View Article : Google Scholar : PubMed/NCBI
47	Arpinati L, Shaul ME, Kaisar-Iluz N, Mali S, Mahroum S and Fridlender ZG: NETosis in cancer: A critical analysis of the impact of cancer on neutrophil extracellular trap (NET) release in lung cancer patients vs. mice Cancer Immunol Immunother. 69:199–213. 2020. View Article : Google Scholar
48	Wu L, Saxena S and Singh RK: Neutrophils in the tumor microenvironment. Adv Exp Med Biol. 1224:1–20. 2020. View Article : Google Scholar : PubMed/NCBI
49	Hisada Y, Grover SP, Maqsood A, Houston R, Ay C, Noubouossie DF, Cooley BC, Wallén H, Key NS, Thålin C, et al: Neutrophils and neutrophil extracellular traps enhance venous thrombosis in mice bearing human pancreatic tumors. Haematologica. 105:218–225. 2020. View Article : Google Scholar :
50	Andzinski L, Kasnitz N, Stahnke S, Wu CF, Gereke M, von Köckritz-Blickwede M, Schilling B, Brandau S, Weiss S and Jablonska J: Type I IFNs induce anti-tumor polarization of tumor associated neutrophils in mice and human. Int J Cancer. 138:1982–1993. 2016. View Article : Google Scholar
51	Natalwala A, Spychal R and Tselepis C: Epithelial-mesenchymal transition mediated tumourigenesis in the gastrointestinal tract. World J Gastroenterol. 14:3792–3797. 2008. View Article : Google Scholar : PubMed/NCBI
52	Wang Y, Chen J, Yang L, Li J, Wu W, Huang M, Lin L and Su S: Tumor-contacted neutrophils promote metastasis by a CD90-TIMP-1 Juxtacrine-paracrine loop. Clin Cancer Res. 25:1957–1969. 2019. View Article : Google Scholar
53	Hu P, Shen M, Zhang P, Zheng C, Pang Z, Zhu L and Du J: Intratumoral neutrophil granulocytes contribute to epithelial-mesenchymal transition in lung adenocarcinoma cells. Tumour Biol. 36:7789–7796. 2015. View Article : Google Scholar : PubMed/NCBI
54	Wu Y, Zhao Q, Peng C, Sun L, Li XF and Kuang DM: Neutrophils promote motility of cancer cells via a hyaluronan-mediatedTLR4/PI3K activation loop. J Pathol. 225:438–447. 2011. View Article : Google Scholar : PubMed/NCBI
55	Zhou Q, Wang X, Yu Z, Wu X, Chen X, Li J, Zhu Z, Liu B and Su L: Transducin (β)-like 1 X-linked receptor 1 promotes gastric cancer progression via the ERK1/2 pathway. Oncogene. 36:1873–1886. 2017. View Article : Google Scholar
56	Molina JR and Adjei AA: The Ras/Raf/MAPK pathway. J Thorac Oncol. 1:7–9. 2006. View Article : Google Scholar
57	Groner B and von Manstein V: Jak Stat signaling and cancer: Opportunities, benefits and side effects of targeted inhibition. Mol Cell Endocrinol. 451:1–14. 2017. View Article : Google Scholar : PubMed/NCBI
58	Lim S and Kaldis P: Cdks, cyclins and CKIs: Roles beyond cell cycle regulation. Development. 140:3079–3093. 2013. View Article : Google Scholar : PubMed/NCBI