Chemokine (C‑C motif) ligand 21/C‑C chemokine receptor type 7 triggers migration and invasion of human lung cancer cells by epithelial‑mesenchymal transition via the extracellular signal‑regulated kinase signaling pathway

Zhong,Guangxin; Chen,Lu; Yin,Ruihong; Qu,Yan; Bao,Yongxing; Xiao,Qiong; Zhang,Zhaolin; Shen,Yaqian; Li,Cailing; Xu,Yun; Zou,Zhigeng; Tian,Hua

doi:10.3892/mmr.2017.6534

Chemokine (C‑C motif) ligand 21/C‑C chemokine receptor type 7 triggers migration and invasion of human lung cancer cells by epithelial‑mesenchymal transition via the extracellular signal‑regulated kinase signaling pathway

Authors:
- Guangxin Zhong
- Lu Chen
- Ruihong Yin
- Yan Qu
- Yongxing Bao
- Qiong Xiao
- Zhaolin Zhang
- Yaqian Shen
- Cailing Li
- Yun Xu
- Zhigeng Zou
- Hua Tian
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Affiliations: Institute of Anatomy and Histology and Embryology, School of Medicine, Shandong University, Jinan, Shandong 250012, P.R. China, Department of Internal Medicine, Jinan First People's Hospital, Jinan, Shandong 250000, P.R. China, Blood Center of General Hospital of Jinan Military Region, Jinan, Shandong 250031, P.R. China, Department of Special Examination, Penglai People's Hospital, Penglai, Shandong 265600, P.R. China, Department of Anatomy, Jining Medical University, Jining, Shandong 272000, P.R. China, Cancer Treatment Center, Shandong Provincial Hospital, Shandong University, Jinan, Shandong 250021, P.R. China
Published online on: May 2, 2017 https://doi.org/10.3892/mmr.2017.6534
Pages: 4100-4108
Copyright: © Zhong et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

C-C chemokine receptor type 7 (CCR7) has been implicated in lymph node metastasis of various cancers. Previous studies have revealed that epithelial‑mesenchymal transition (EMT) is involved in the chemotactic process mediated by CCR7 and its ligands in various types of carcinoma. However, the underlying mechanism of this process remains to be fully elucidated. The present study investigated whether chemokine (C‑C motif) ligand 21 (CCL21)/CCR7 may activate EMT of lung cancer cells and their associated signaling pathways. A549 and H520 lung cancer cell lines were examined in vitro in the present study. The results indicated that A549 and H520 expressed CCR7, but reduced levels of CCL21. Following stimulation of lung cancer cell lines with CCL21, the expression of the epithelial marker E‑cadherin was downregulated, and the mesenchymal markers Vimentin/Slug and extracellular signal‑regulated kinase (ERK) were upregulated. In addition, the ERK inhibitor PD98059 may inhibit EMT caused by CCL21, and decreased cell migration and invasion initiated by CCL21. Furthermore, lung adenocarcinoma tissues from 50 patients who underwent lung cancer operations were investigated by immunohistochemistry. The findings revealed that CCR7, Slug and Vimentin were highly expressed in lung carcinoma tissues, and were significantly associated with lymph node metastasis and clinical pathological stages, respectively. CCR7 expression was correlated positively with expression levels of Slug and Vimentin. CCL21 was expressed positively in the endothelium of lymphatic vessels adjacent to cancer cells, and weakly in lung cancer cells. Collectively, these results demonstrated that CCL21/CCR7 may activate EMT in lung cancer cells via the ERK1/2 signaling pathway. The current study provides evidence that a close interaction exists between CCL21/CCR7chemotaxis and EMT procedures in lung cancer metastasis, providing a basis for the development of therapeutic targets.

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1	Thiery JP, Acloque H, Huang RY and Nieto MA: Epithelial-mesenchymal transitions in development and disease. Cell. 139:871–890. 2009. View Article : Google Scholar
2	De Craene B and Berx G: Regulatory networks defining EMT during cancer initiation and progression. Nat Rev Cancer. 13:97–110. 2013. View Article : Google Scholar
3	Mukaida N and Baba T: Chemokines in tumor development and progression. Exp Cell Res. 318:95–102. 2012. View Article : Google Scholar
4	Zlotnik A, Burkhardt AM and Homey B: Homeostatic chemokine receptors and organ-specific metastasis. Nat Rev Immunol. 11:597–606. 2011. View Article : Google Scholar
5	Förster R, Davalos-Misslitz AC and Rot A: CCR7 and its ligands: Balancing immunity and tolerance. Nat Rev Immunol. 8:362–371. 2008. View Article : Google Scholar
6	Carlsen HS, Haraldsen G, Brandtzaeg P and Baekkevold ES: Disparate lymphoid chemokine expression in mice and men: No evidence of CCL21 synthesis by human high endothelial venules. Blood. 106:444–446. 2005. View Article : Google Scholar
7	Baekkevold ES, Yamanaka T, Palframan RT, Carlsen HS, Reinholt FP, von Andrian UH, Brandtzaeg P and Haraldsen G: The CCR7 ligand ELC (CCL19) is transcytosed in high endothelial venules and mediates T cell recruitment. J Exp Med. 193:1105–1112. 2001. View Article : Google Scholar :
8	Gunn MD, Tangemann K, Tam C, Cyster JG, Rosen SD and Williams LT: A chemokine expressed in lymphoid high endothelial venules promotes the adhesion and chemotaxis of naive T lymphocytes. Proc Natl Acad Sci USA. 95:pp. 258–263. 1998; View Article : Google Scholar :
9	Sperveslage J, Frank S, Heneweer C, Egberts J, Schniewind B, Buchholz M, Bergmann F, Giese N, Munding J, Hahn SA, et al: Lack of CCR7 expression is rate limiting for lymphatic spread of pancreatic ductal adenocarcinoma. Int J Cancer. 131:E371–E381. 2012. View Article : Google Scholar
10	Legler DF, Uetz-von Allmen E and Hauser MA: CCR7: Roles in cancer cell dissemination, migration and metastasis formation. Int J Biochem Cell Biol. 54:78–82. 2014. View Article : Google Scholar
11	Li X, Ma Q, Xu Q, Liu H, Lei J, Duan W, Bhat K, Wang F, Wu E and Wang Z: SDF-1/CXCR4 signaling induces pancreatic cancer cell invasion and epithelial-mesenchymal transition in vitro through non-canonical activation of Hedgehog pathway. Cancer Lett. 322:169–176. 2012. View Article : Google Scholar :
12	Chen K, Li Z, Jiang P, Zhang X, Zhang Y, Jiang Y, He Y and Li X: Co-expression of CD133, CD44v6 and human tissue factor is associated with metastasis and poor prognosis in pancreatic carcinoma. Oncol Rep. 32:755–763. 2014.
13	Lee SH, Kang HY, Kim KS, Nam BY, Paeng J, Kim S, Li JJ, Park JT, Kim DK, Han SH, et al: The monocyte chemoattractant protein-1 (MCP-1)/CCR2 system is involved in peritoneal dialysis-related epithelial-mesenchymal transition of peritoneal mesothelial cells. Lab Invest. 92:1698–1711. 2012. View Article : Google Scholar
14	Marsigliante S, Vetrugno C and Muscella A: Paracrine CCL20 loop induces epithelial-mesenchymal transition in breast epithelial cells. Mol Carcinog. 55:1175–1186. 2016. View Article : Google Scholar
15	Li F, Zou Z, Suo N, Zhang Z, Wan F, Zhong G, Qu Y, Ntaka KS and Tian H: CCL21/CCR7 axis activating chemotaxis accompanied with epithelial-mesenchymal transition in human breast carcinoma. Med Oncol. 31:1802014. View Article : Google Scholar
16	Zhang J, Zhou Y and Yang Y: CCR7 pathway induces epithelial-mesenchymal transition through up-regulation of Snail signaling in gastric cancer. Med Oncol. 32:4672015.
17	Siegel RL, Miller KD and Jemal A: Cancer Statistics, 2017. CA Cancer J Clin. 67:7–30. 2017. View Article : Google Scholar
18	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
19	Goldstraw P, Chansky K, Crowley J, Rami-Porta R, Asamura H, Eberhardt WE, Nicholson AG, Groome P, Mitchell A, Bolejack V, et al: The IASLC lung cancer staging project: proposals for revision of the TNM stage groupings in the forthcoming (Eighth) edition of the TNM classification for lung cancer. J Thorac Oncol. 11:39–51. 2016. View Article : Google Scholar
20	Chen X, Ye S, Xiao W, Wang W, Luo L and Liu Y: ERK1/2 pathway mediates epithelial-mesenchymal transition by cross-interacting with TGFβ/Smad and Jagged/Notch signaling pathways in lens epithelial cells. Int J Mol Med. 33:1664–1670. 2014.
21	Buonato JM and Lazzara MJ: ERK1/2 blockade prevents epithelial-mesenchymal transition in lung cancer cells and promotes their sensitivity to EGFR inhibition. Cancer Res. 74:309–319. 2014. View Article : Google Scholar
22	Maekawa S, Iwasaki A, Shirakusa T, Kawakami T, Yanagisawa J, Tanaka T, Shibaguchi H, Kinugasa T and Kuroki M and Kuroki M: Association between the expression of chemokine receptors CCR7 and CXCR3, and lymph node metastatic potential in lung adenocarcinoma. Oncol Rep. 19:1461–1468. 2008.
23	Shields JD, Fleury ME, Yong C, Tomei AA, Randolph GJ and Swartz MA: Autologous chemotaxis as a mechanism of tumor cell homing to lymphatics via interstitial flow and autocrine CCR7 signaling. Cancer Cell. 11:526–538. 2007. View Article : Google Scholar
24	Li Y, Zhang Q, Jiang L, Qiu X and Wang E: Upregulation of the Chemokine Receptor CCR7 expression by HIF-1alpha and HIF-2alpha in non-small cell lung cancer. Zhongguo Fei Ai Za Zhi. 11:724–728. 2008.(In Chinese).
25	Li Y, Zhang Q, Wang Y, Qiu X and Wang E: Effects of hypoxia on the expression of CCR7 and proliferation, invasiveness of A549 cells. Zhongguo Fei Ai Za Zhi. 11:704–706. 2008.(In Chinese).
26	Pang MF, Georgoudaki AM, Lambut L, Johansson J, Tabor V, Hagikura K, Jin Y, Jansson M, Alexander JS, Nelson CM, et al: TGF-β1-induced EMT promotes targeted migration of breast cancer cells through the lymphatic system by the activation of CCR7/CCL21-mediated chemotaxis. Oncogene. 35:748–760. 2016. View Article : Google Scholar
27	Papageorgis P: TGFβ signaling in tumor initiation, epithelial-to-mesenchymal transition and metastasis. J Oncol. 2015:5871932015. View Article : Google Scholar :