EPCR promotes MGC803 human gastric cancer cell tumor angiogenesis in vitro through activating ERK1/2 and AKT in a PAR1‑dependent manner

Wang,Qingling; Tang,Yangyang; Wang,Tianyuan; Yang,Hong‑Li; Wang,Xinyue; Ma,Hongru; Zhang,Peng

doi:10.3892/ol.2018.8869

August-2018 Volume 16 Issue 2

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August-2018 Volume 16 Issue 2

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Article Open Access

EPCR promotes MGC803 human gastric cancer cell tumor angiogenesis in vitro through activating ERK1/2 and AKT in a PAR1‑dependent manner

Authors:
- Qingling Wang
- Yangyang Tang
- Tianyuan Wang
- Hong‑Li Yang
- Xinyue Wang
- Hongru Ma
- Peng Zhang
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Affiliations: Department of Pathology, Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China, School of Anesthesiology, Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China, School of Clinical Medicine, Xuzhou Medical University, Xuzhou, Jiangsu 221004, P.R. China, Jiangsu Key Laboratory of Immunity and Metabolism, Xuzhou, Jiangsu 221004, P.R. China

Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 1565-1570
|
Published online on: June 1, 2018

https://doi.org/10.3892/ol.2018.8869
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Abstract

The endothelial cell protein C receptor (EPCR) serves a key role in activated protein C (APC)‑mediated cytoprotective effects in endothelial cells, and is involved in the development of certain types of human cancer. To the best of our knowledge, the present study is the first to demonstrate that EPCR may exert effects on gastric cancer angiogenesis in vitro. To detect microvessel density (MVD), the microvascular endothelial cells were stained for cluster of differentiation (CD)31 and CD34 in 61 cases of surgical resection of gastric carcinoma tissues, and the association between the expression of EPCR protein and MVD was analyzed. In addition, to analyze the effect of EPCR expressed by gastric cancer cells on the proliferation, migration and angiogenic abilities of endothelial cells, human umbilical vein endothelial cells (HUVECs) were cultured with tumor‑conditioned medium derived from EPCR knockdown or protease‑activated receptor 1 (PAR1)‑blocked MGC803 gastric cancer cells. A CCK‑8 assay was used to assess the proliferation ability of the HUVECs. A Transwell assay was performed to assess the migration ability of the HUVECs and a Matrigel‑based tube formation assay was used to assess the angiogenic activity of the HUVECs. The results demonstrated that the expression of EPCR was correlated with the MVD of gastric cancer tissues. When cultured with tumor‑conditioned medium derived from EPCR knockdown or PAR1‑blocked MGC803 cells, the proliferation, migration and tubules formation abilities of HUVECs were markedly inhibited markedly. The expression of phosphorylated (p)‑extracellular signal regulated kinase 1/2, p‑protein kinase B (AKT; s473) and p‑AKT (T308) in the HUVECs was decreased. In addition, EPCR knockdown inhibited PAR1 activation in the MGC803 cells. These results indicated that the expression of EPCR in gastric cancer cell line MGC803 contributes to tumor angiogenesis in vitro by activating ERK1/2 and AKT, and that this effect of EPCR is dependent on PAR1 activation.

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1	Liu CC, Shen Z, Kung HF and Lin MC: Cancer gene therapy targeting angiogenesis: An updated review. World J Gastroenterol. 12:6941–6948. 2006. View Article : Google Scholar : PubMed/NCBI
2	Thiyagarajan M, Cheng T and Zlokovic BV: Endothelial cell protein C receptor: Role beyond endothelium? Circ Res. 100:155–157. 2007. View Article : Google Scholar : PubMed/NCBI
3	Crawley JT: Multiple roles of the endothelial cell protein C receptor. J Thromb Haemost. 5:1813–1816. 2007. View Article : Google Scholar : PubMed/NCBI
4	Mohan Rao LV, Esmon CT and Pendurthi UR: Endothelial cell protein C receptor: A multiliganded and multifunctional receptor. Blood. 124:1553–1562. 2014. View Article : Google Scholar : PubMed/NCBI
5	Riewald M, Petrovan RJ, Donner A and Ruf W: Activated protein C signals through the thrombin receptor PAR1 in endothelial cells. J Endotoxin Res. 9:317–321. 2003. View Article : Google Scholar : PubMed/NCBI
6	Uchiba M, Okajima K, Oike Y, Ito Y, Fukudome K, Isobe H and Suda T: Activated protein C induces endothelial cell proliferation by mitogen-activated protein kinase activation in vitro and angiogenesis in vivo. Circ Res. 95:34–41. 2004. View Article : Google Scholar : PubMed/NCBI
7	Shua F, Kobayashia H, Fukudomeb K, Tsuneyoshib N, Kimotob M and Teraoa T: Activated protein C suppresses tissue factor expression on U937 cells in the endothelial protein C receptor-dependent manner. FEBS Lett. 477:208–212. 2000. View Article : Google Scholar : PubMed/NCBI
8	Keshava S, Sahoo S, Tucker TA, Idell S, Rao LV and Pendurthi UR: Endothelial cell protein C receptor opposes mesothelioma growth driven by tissue factor. Cancer Res. 73:3963–3973. 2013. View Article : Google Scholar : PubMed/NCBI
9	Ducros E, Mirshahi S, Azzazene D, Camilleri-Broët S, Mery E, Al Farsi H, Althawadi H, Besbess S, Chidiac J, Pujade-Lauraine E, et al: Endothelial protein C receptor expressed by ovarian cancer cells as a possible biomarker of cancer onset. Int J Oncol. 41:433–440. 2012. View Article : Google Scholar : PubMed/NCBI
10	Antón I, Molina E, Luis-Ravelo D, Zandueta C, Valencia K, Ormazabal C, Martínez-Canarias S, Perurena N, Pajares MJ, Agorreta J, et al: Receptor of activated protein C promotes metastasis and correlates with clinical outcome in lung adenocarcinoma. Am J Respir Crit Care Med. 186:96–105. 2012. View Article : Google Scholar : PubMed/NCBI
11	Heng W, Mu CY, Chen C, Huang JA and Wang ZY: Endothelial cell protein C receptor (EPCR) is expressed by lung carcinoma and correlated with clinical parameters. Clin Lab. 59:375–380. 2013. View Article : Google Scholar : PubMed/NCBI
12	Gramling MW, Beaulieu LM and Church FC: Activated protein C enhances cell motility of endothelial cells and MDA-MB-231 breast cancer cells by intracellular signal transduction. Exp Cell Res. 316:314–328. 2010. View Article : Google Scholar : PubMed/NCBI
13	Perurena N, Zandueta C, Martínez-Canarias S, Moreno H, Vicent S, Almeida AS, Guruceaga E, Gomis RR, Santisteban M, Egeblad M, et al: EPCR promotes breast cancer progression by altering SPOCK1/testican 1-mediated 3D growth. J Hematol Oncol. 10:232017. View Article : Google Scholar : PubMed/NCBI
14	Wang Q, Liu Q, Wang T, Yang H, Han Z and Zhang P: Endothelial cell protein C receptor promotes MGC803 gastric cancer cells proliferation and migration by activating ERK1/2. Med Oncol. 32:1622015. View Article : Google Scholar : PubMed/NCBI
15	Jang HK, Kin BS, Han J, Yoon JK, Lee JR, Jeong GJ and Shin JY: Therapeutic angiogenesis using tumor cell-conditioned medium. Biotechnol Prog. 32:456–464. 2016. View Article : Google Scholar : PubMed/NCBI
16	Xu LN, Xu BN, Cai J, Yang JB and Lin N: Tumor-associated fibroblast-conditioned medium promotes tumor cell proliferation and angiogenesis. Genet Mol Res. 12:5863–5871. 2013. View Article : Google Scholar : PubMed/NCBI
17	Zhang T and Jiang CL: Tumor conditioned medium regulates the proliferation, adhesion and migration of human umbilical vein endothelial cells. Sheng Li Xue Bao. 63:256–260. 2011.(In Chinese). PubMed/NCBI
18	Liu T, Jabbes M, Nedrow-Byers JR, Wu LY, Bryan JN and Berkman CE: Detection of prostate-specific membrane antigen on HUVECs in response to breast tumor-conditioned medium. Int J Oncol. 38:1349–1355. 2011.PubMed/NCBI
19	Koizumi S, Gu C, Amano S, Yamamoto S, Ihara H, Tokuyama T and Namba H: Migration of mouse-induced pluripotent stem cells to glioma-conditioned medium is mediated by tumor-associated specific growth factors. Oncol Lett. 2:283–288. 2011. View Article : Google Scholar : PubMed/NCBI
20	Peng Y, Li J and Geng M: The glycan profile of endothelial cells in the present of tumor-conditioned medium and potential roles of beta-1,6-GlcNAc branching on HUVEC conformation. Mol Cell Biochem. 340:143–152. 2010. View Article : Google Scholar : PubMed/NCBI
21	Falahat R, Wiranowska M, Gallant ND, Toomey R, Hill R and Alcantar N: A cell ELISA for the quantification of MUC1 mucin (CD227) expressed by cancer cells of epithelial and neuroectodermal origin. Cell Immunol. 298:96–103. 2015. View Article : Google Scholar : PubMed/NCBI
22	Niessen F, Furlan-Freguia C, Fernández JA, Mosnier LO, Castellino FJ, Weiler H, Rosen H, Griffin JH and Ruf W: Endogenous EPCR/aPC-PAR1 signaling prevents inflammation-induced vascular leakage and lethality. Blood. 113:2859–2866. 2009. View Article : Google Scholar : PubMed/NCBI
23	Sundaram J, Keshava S, Gopalakrishnan R, Esmon CT, Pendurthi UR and Rao LV: Factor VIIa binding to endothelial cell protein C receptor protects vascular barrier integrity in vivo. J Thromb Haemost. 12:690–700. 2014. View Article : Google Scholar : PubMed/NCBI
24	Mosnier LO and Griffin JH: Inhibition of staurosporine-induced apoptosis of endothelial cells by activated protein C requires protease-activated receptor-1 and endothelial cell protein C receptor. Biochem J. 373:65–70. 2003. View Article : Google Scholar : PubMed/NCBI
25	Hun Lee J, Won S and Stein DG: Progesterone attenuates thrombin-induced endothelial barrier disruption in the brain endothelial cell line bEnd.3: The role of tight junction proteins and the endothelial protein C receptor. Brain Res. 1613:73–80. 2015. View Article : Google Scholar : PubMed/NCBI
26	Sen P, Sahoo S, Pendurthi UR and Rao LV: Zinc modulates the interaction of protein C and activated protein C with endothelial cell protein C receptor. J Biol Chem. 285:20410–20420. 2010. View Article : Google Scholar : PubMed/NCBI