Increase in CD4+FOXP3+ regulatory T cell number and upregulation of the HGF/c‑Met signaling pathway during the liver metastasis of colorectal cancer

Huang,Xiaoming; Chen,Zexian; Zhang,Nanrong; Zhu,Caiyan; Lin,Xutao; Yu,Jiandong; Chen,Zhiping; Lan,Ping; Wan,Yunle

doi:10.3892/ol.2020.11785

Increase in CD4+FOXP3+ regulatory T cell number and upregulation of the HGF/c‑Met signaling pathway during the liver metastasis of colorectal cancer

Authors:
- Xiaoming Huang
- Zexian Chen
- Nanrong Zhang
- Caiyan Zhu
- Xutao Lin
- Jiandong Yu
- Zhiping Chen
- Ping Lan
- Yunle Wan
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Affiliations: Department of Hepatobiliary Surgery, The Sixth Affiliated Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510655, P.R. China, Guangdong Provincial Key Laboratory of Colorectal and Pelvic Floor Diseases, The Sixth Affiliated Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510655, P.R. China, Department of Anesthesiology, The Sixth Affiliated Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510655, P.R. China, Department of Pharmacy, The Sixth Affiliated Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510655, P.R. China, Department of Endoscopy Center, The Sixth Affiliated Hospital, Sun Yat‑Sen University, Guangzhou, Guangdong 510655, P.R. China
Published online on: June 26, 2020 https://doi.org/10.3892/ol.2020.11785
Pages: 2113-2118
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Colorectal cancer (CRC) is the third and second most common type of cancer diagnosed in males and females, respectively, and is the fourth leading cause of cancer‑associated mortality worldwide. Liver metastasis is the primary cause of mortality in patients with CRC, and therefore requires therapeutic focus. Regulatory T cells (Tregs) and hepatic stellate cells (HSCs) are potentially involved in regulating the immune response during liver metastasis. The aim of the present study was to evaluate the influence of CD4+ forkhead box p3 (Foxp3)+ Tregs and the HGF/c‑Met signaling pathway in the liver metastasis of CRC. A model of the latter was established using Balb/c mice via splenic injection of human CRC cells (CT‑26 line). The mice were monitored for 3 weeks after being injected, and the spleens and livers were removed on day 22 for further analysis. Moreover, the single‑cell suspensions were labeled with CD4 and Foxp3 antibodies, and were analyzed using flow cytometry. Expression levels of α‑smooth muscle actin (SMA), hepatocyte growth factor (HGF) and hepatocyte growth factor receptor (c‑Met) were analyzed using immunohistochemistry. Mice injected with CT‑26 cells exhibited signs of illness and significant weight loss, compared with the control mice (P=0.013), and they also developed liver metastases, at an average of 20.5 tumors per mouse. Pathological evaluation using hematoxylin and eosin staining confirmed the tumors as liver metastases of CRC. The numbers of CD4+ T cells were significantly decreased in the spleen (P<0.001) and liver (P=0.003) of tumor‑bearing mice, while the proportions of CD4+FOXP3+ Tregs increased significantly in the spleen (P<0.001) and liver (P=0.026) compared with that in the controls. Additionally, α‑SMA, HGF and c‑Met levels increased significantly during metastatic growth in the liver. In conclusion, CD4+FOXP3+ Treg levels increased and the HGF/c‑Met pathway was upregulated during the liver metastasis of CRC in mice, indicating the presence of potential therapeutic targets.

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1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Bengmark S and Hafstrom L: The natural history of primary and secondary malignant tumors of the liver. I. The prognosis for patients with hepatic metastases from colonic and rectal carcinoma by laparotomy. Cancer Am Cancer Soc. 23:198–202. 1969.
3	Manfredi S, Lepage C, Hatem C, Coatmeur O, Faivre J and Bouvier AM: Epidemiology and management of liver metastases from colorectal cancer. Ann Surg. 244:254–259. 2006. View Article : Google Scholar : PubMed/NCBI
4	Konopke R, Roth J, Volk A, Pistorius S, Folprecht G, Zöphel K, Schuetze C, Laniado M, Saeger HD and Kersting S: Colorectal liver metastases: An update on palliative treatment options. J Gastrointestin Liver Dis. 21:83–91. 2012.PubMed/NCBI
5	Lykoudis PM, O'Reilly D, Nastos K and Fusai G: Systematic review of surgical management of synchronous colorectal liver metastases. Br J Surg. 101:605–612. 2014. View Article : Google Scholar : PubMed/NCBI
6	Robinson MW, Harmon C and O'Farrelly C: Liver immunology and its role in inflammation and homeostasis. Cell Mol Immunol. 13:267–276. 2016. View Article : Google Scholar : PubMed/NCBI
7	Nishikawa H and Sakaguchi S: Regulatory T cells in cancer immunotherapy. Curr Opin Immunol. 27:1–7. 2014. View Article : Google Scholar : PubMed/NCBI
8	Huang X, Zou Y, Lian L, Wu X, He X, He X, Wu X, Huang Y and Lan P: Changes of T cells and cytokines TGF-β1 and IL-10 in mice during liver metastasis of colon carcinoma: Implications for liver anti-tumor immunity. J Gastrointest Surg. 17:1283–1291. 2013. View Article : Google Scholar : PubMed/NCBI
9	Bárcena C, Stefanovic M, Tutusaus A, Martinez-Nieto GA, Martinez L, García-Ruiz C, de Mingo A, Caballeria J, Fernandez-Checa JC, Marí M and Morales A: Angiogenin secretion from hepatoma cells activates hepatic stellate cells to amplify a self-sustained cycle promoting liver cancer. Sci Rep. 5:79162015. View Article : Google Scholar : PubMed/NCBI
10	Gupta G, Khadem F and Uzonna JE: Role of hepatic stellate cell (HSC)-derived cytokines in hepatic inflammation and immunity. Cytokine. 124:1545422019. View Article : Google Scholar : PubMed/NCBI
11	Lu DH, Guo XY, Qin SY, Luo W, Huang XL, Chen M, Wang JX, Ma SJ, Yang XW and Jiang HX: Interleukin-22 ameliorates liver fibrogenesis by attenuating hepatic stellate cell activation and downregulating the levels of inflammatory cytokines. World J Gastroenterol. 21:1531–1545. 2015. View Article : Google Scholar : PubMed/NCBI
12	Weiskirchen R and Tacke F: Cellular and molecular functions of hepatic stellate cells in inflammatory responses and liver immunology. Hepatobiliary Surg Nutr. 3:344–363. 2014.PubMed/NCBI
13	Patel MB, Pothula SP, Xu Z, Lee AK, Goldstein D, Pirola RC, Apte MV and Wilson JS: The role of the hepatocyte growth factor/c-MET pathway in pancreatic stellate cell-endothelial cell interactions: Antiangiogenic implications in pancreatic cancer. Carcinogenesis. 35:1891–1900. 2014. View Article : Google Scholar : PubMed/NCBI
14	Cai W, Rook SL, Jiang ZY, Takahara N and Aiello LP: Mechanisms of hepatocyte growth factor-induced retinal endothelial cell migration and growth. Invest Ophthalmol Vis Sci. 41:1885–1893. 2000.PubMed/NCBI
15	Pothula SP, Xu Z, Goldstein D, Biankin AV, Pirola RC, Wilson JS and Apte MV: Hepatocyte growth factor inhibition: A novel therapeutic approach in pancreatic cancer. Br J Cancer. 114:269–280. 2016. View Article : Google Scholar : PubMed/NCBI
16	Parr C and Jiang WG: Expression of hepatocyte growth factor/scatter factor, its activator, inhibitors and the c-Met receptor in human cancer cells. Int J Oncol. 19:857–863. 2001.PubMed/NCBI
17	Syed Khaja A, Toor SM, El SH, Ali BR and Elkord E: Intratumoral FoxP3⁺Helios⁺ regulatory T cells upregulating immunosuppressive molecules are expanded in human colorectal cancer. Front Immunol. 8:6192017. View Article : Google Scholar : PubMed/NCBI
18	Liu HY, Huang ZL, Yang GH, Lu WQ and Yu NR: Inhibitory effect of modified citrus pectin on liver metastases in a mouse colon cancer model. World J Gastroenterol. 14:7386–7391. 2008. View Article : Google Scholar : PubMed/NCBI
19	Takeuchi Y and Nishikawa H: Roles of regulatory T cells in cancer immunity. Int Immunol. 28:401–409. 2016. View Article : Google Scholar : PubMed/NCBI
20	Fontenot JD, Gavin MA and Rudensky AY: Foxp3 programs the development and function of CD4⁺CD25⁺ regulatory T cells. Nat Immunol. 4:330–336. 2003. View Article : Google Scholar : PubMed/NCBI
21	Shimizu J, Yamazaki S and Sakaguchi S: Induction of tumor immunity by removing CD25⁺CD4⁺ T cells: A common basis between tumor immunity and autoimmunity. J Immunol. 163:5211–5218. 1999.PubMed/NCBI
22	Onizuka S, Tawara I, Shimizu J, Sakaguchi S, Fujita T and Nakayama E: Tumor rejection by in vivo administration of anti-CD25 (interleukin-2 receptor alpha) monoclonal antibody. Cancer Res. 59:3128–3133. 1999.PubMed/NCBI
23	Almeida AR, Ciernik IF, Sallusto F and Lanzavecchia A: CD4⁺ CD25⁺ Treg regulate the contribution of CD8⁺ T-cell subsets in repopulation of the lymphopenic environment. Eur J Immunol. 40:3478–3488. 2010. View Article : Google Scholar : PubMed/NCBI
24	Zanin-Zhorov A, Ding Y, Kumari S, Attur M, Hippen KL, Brown M, Blazar BR, Abramson SB, Lafaille JJ and Dustin ML: Protein kinase C-theta mediates negative feedback on regulatory T cell function. Science. 328:372–376. 2010. View Article : Google Scholar : PubMed/NCBI
25	Arlt F and Stein U: Colon cancer metastasis: MACC1 and Met as metastatic pacemakers. Int J Biochem Cell Biol. 41:2356–2359. 2009. View Article : Google Scholar : PubMed/NCBI
26	Bradley CA, Salto-Tellez M, Laurent-Puig P, Bardelli A, Rolfo C, Tabernero J, Khawaja HA, Lawler M, Johnston PG and Van Schaeybroeck S; MErCuRIC consortium, : Targeting c-MET in gastrointestinal tumours: Rationale, opportunities and challenges. Nat Rev Clin Oncol. 14:562–576. 2017. View Article : Google Scholar : PubMed/NCBI
27	Shojaei F, Simmons BH, Lee JH, Lappin PB and Christensen JG: HGF/c-Met pathway is one of the mediators of sunitinib-induced tumor cell type-dependent metastasis. Cancer Lett. 320:48–55. 2012. View Article : Google Scholar : PubMed/NCBI
28	Osada S, Matsui S, Komori S, Yamada J, Sanada Y, Ihawa A, Tanaka Y, Tokuyama Y, Okumura N, Nonaka K, et al: Effect of hepatocyte growth factor on progression of liver metastasis in colorectal cancer. Hepatogastroenterology. 57:76–80. 2010.PubMed/NCBI
29	Olguín JE, Medina-Andrade I, Molina E, Vázquez A, Pacheco-Fernández T, Saavedra R, Pérez-Plasencia C, Chirino YI, Vaca-Paniagua F, Arias-Romero LE, et al: Early and partial reduction in CD4⁺Foxp3⁺ regulatory T cells during Colitis-associated colon cancer induces CD4⁺ and CD8⁺ T cell activation inhibiting tumorigenesis. J Cancer. 9:239–249. 2018. View Article : Google Scholar : PubMed/NCBI