Effects of silenced PAR-2 on cell proliferation, invasion and metastasis of esophageal cancer

Chen,Jinmei; Xie,Liqun; Zheng,Yanmin; Liu,Caihong

doi:10.3892/ol.2017.6711

Effects of silenced PAR-2 on cell proliferation, invasion and metastasis of esophageal cancer

Authors:
- Jinmei Chen
- Liqun Xie
- Yanmin Zheng
- Caihong Liu
View Affiliations

Affiliations: Department of Gastroenterology, Affiliated Hospital of Logistics University of People's Armed Police Force, Tianjin 300162, P.R. China
Published online on: August 3, 2017 https://doi.org/10.3892/ol.2017.6711
Pages: 4115-4121
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The present study aimed to investigate the effect of protease-activated receptor 2 (PAR‑2) on cell proliferation, invasion and metastasis in the esophageal EC109 cell line. Two short hairpin RNA (shRNA) expression plasmids were constructed based on the PAR‑2 mRNA sequence in humans, and they were transfected into the EC109 esophageal cancer cell line, and the stable interference cell line (shRNA‑PAR‑2 EC109) was obtained by puromycin selection. Following transfection of PAR‑2 shRNA‑1, PAR‑2 expression was significantly downregulated in mRNA level and protein level in EC109 cells (P<0.05). The proliferation of EC109 cells transfected with PAR‑2 shRNA was significantly lower than the negative control group (P<0.05). At 24, 48 and 72 h, the ratio of proliferation inhibition was 15.92, 24.89 and 32.28%, respectively. Compared with the control group, S‑phase arrest was observed in cells transfected with shRNA‑PAR‑2. The ratio of cells in the S phase was 32.79±4.06, 26.54±1.37 and 33.45±2.46% at 24, 48 and 72 h, respectively. For invasion, the number of invasive cells was significantly lower in shRNA‑PAR2‑2 cells compared with the control group (P<0.05). For metastasis assay, the number of invasive cells was significantly lower in shRNA‑PAR2‑2 cells compared with the control group (P<0.01). In the present study, the PAR‑2 shRNA plasmid was constructed successfully, which can significantly downregulate PAR‑2 expression in EC109 cells. Subsequent to silencing of PAR‑2, the proliferation of EC109 cells was inhibited and the capabilities of invasion and migration were reduced. It is indicated that PAR‑2 may be a potential target in esophageal cancer.

View Figures

View References

1	Montgomery EA: Oesophageal cancerWorld Cancer Report 2014. Stewart BW and Wild CP: IARC; Lyon: pp. 374–382. 2014
2	Hao J and Shao K: The epidemiology, current status of management, challenge and future strategy for esophageal cancer. Chin Oncol. 21:501–504. 2011.
3	Ciocirlan M, Lapalus MG, Hervieu V, Souquet JC, Napoléon B, Scoazec JY, Lefort C, Saurin JC and Ponchon T: Endoscopic mucosal resection for squamous premalignant and early malignant lesions of the esophagus. Endoscopy. 39:24–29. 2007. View Article : Google Scholar : PubMed/NCBI
4	Kim T, Grobmyer SR, Smith R, Ben-David K, Ang D, Vogel SB and Hochwald SN: Esophageal cancer-the five year survivors. J Surg Oncol. 103:179–183. 2011. View Article : Google Scholar : PubMed/NCBI
5	Ossovskaya VS and Bunnett NW: Protease-activated receptors: Contribution to physiology and disease. Physiol Rev. 84:579–621. 2004. View Article : Google Scholar : PubMed/NCBI
6	Caruso R, Pallone F, Fina D, Gioia V, Peluso I, Caprioli F, Stolfi C, Perfetti A, Spagnoli LG, Palmieri G, et al: Protease-activated receptor-2 activation in gastric cancer cells promotes epidermal growth factor receptor transactivation and proliferation. Am J Pathol. 169:268–278. 2006. View Article : Google Scholar : PubMed/NCBI
7	Shimamoto R, Sawada T, Uchima Y, Inoue M, Kimura K, Yamashita Y, Yamada N, Nishihara T, Ohira M and Hirakawa K: A role for protease-activated receptor-2 in pancreatic cancer cell proliferation. Int J Oncol. 24:1401–1406. 2004.PubMed/NCBI
8	Zhou J, Xie LQ, Li X, Chen X, Chen L, Zheng Y and Li F: Protease-activated receptor-2 agonists promote cell invasion and metastasis in human esophageal cancer cell line EC109. World Chin J Digestol. 13:1313–1319. 2010.(In Chinese).
9	Macfarlane SR, Seatter MJ, Kanke T, Hunter GD and Plevin R: Proteinase-activated receptors. Pharmacol Rev. 53:245–282. 2001.PubMed/NCBI
10	Nystedt S, Emilsson K, Wahlestedt C and Sundelin J: Molecular cloning of a potential proteinase activated receptor. Proc Natl Acad Sci USA. 91:9208–9212. 1994. View Article : Google Scholar : PubMed/NCBI
11	Inci K, Edebo A, Olbe L and Casselbrant A: Expression of protease-activated-receptor 2 (PAR-2) in human esophageal mucosa. Scand J Gastroenterol. 44:664–671. 2009. View Article : Google Scholar : PubMed/NCBI
12	Uchima Y, Sawada T and Hirakawa K: Action of antiproteases on pancreatic cancer cells. JOP. 8:(4 Suppl). S479–S487. 2007.
13	Uusitalo-Jarvinen H, Kurokawa T, Mueller BM, Andrade-Gordon P, Friedlander M and Ruf W: Role of protease activated receptor 1 and 2 signaling in hypoxia induced angiogenesis. Arterioscler Thromb Vasc Biol. 27:1456–1462. 2007. View Article : Google Scholar : PubMed/NCBI
14	Awasthi V, Mandal SK, Papanna V, Rao LV and Pendurthi UR: Modulation of tissue factor VIIa signaling by lipid rafts and caveolae. Arterioscler Thromb Vasc Biol. 27:1447–1455. 2007. View Article : Google Scholar : PubMed/NCBI
15	Fujimoto D, Hirono Y, Goi T, Katayama K, Hirose K and Yamaguchi A: Expression of protease activated receptor-2 (PAR-2) in gastric cancer. J Surg Oncol. 93:139–144. 2006. View Article : Google Scholar : PubMed/NCBI
16	Ribeiro FS, Simão TA, Amoêdo ND, Andreollo NA, Lopes LR, Acatauassu R, Rumjanek FD, Albano RM, Pinto LF and Monteiro RQ: Evidence for increased expression of tissue factor and protease-activated receptor-1 in human esophageal cancer. Oncol Rep. 21:1599–1604. 2009.PubMed/NCBI
17	Hannon GJ: RNA interference. Nature. 418:244–251. 2002. View Article : Google Scholar : PubMed/NCBI
18	Brusselmans K, De Schrijver E, Verhoeven G and Swinnen JV: RNA interference-mediated silencing of the acetyl-CoA-carboxylase-alpha gene induces growth inhibition and apoptosis of prostate cancer cells. Cancer Res. 65:6719–6725. 2005. View Article : Google Scholar : PubMed/NCBI
19	Filipowicz W: RNAI: The nuts and bolts of the RISC machine. Cell. 122:17–20. 2005. View Article : Google Scholar : PubMed/NCBI
20	Mette MF, Aufsatz W, Kanno T, Daxinger L, Rovina P, Matzke M and Matzke AJ: Analysis of double-strand RNA and small RNAs involved in RNA-mediated transcriptional gene silencing. Methods Mol Biol. 309:61–82. 2005.PubMed/NCBI
21	Zhang H, Zheng X, Chen K, et al: SiRNA inhibit the expression of VEGF-C in patients with esophageal carcinoma. Cancer Res Prev Treatment. 37:132–135. 2010.(In Chinese).
22	Zheng YM, Xie LQ, Zhao JY, Li X, Chen XY, Chen L, Hai O and Zhou J: Effects of proteinase activated receptor-2 agonists on proliferation of hepatoma cells. Zhonghua Gan Zang Bing Za Zhi. 17:701–702. 2009.(In Chinese). PubMed/NCBI
23	Zeng ZS, Shu WP, Cohen AM and Guillem JG: Matrix metalloproteinase-7 expression in colorectal cancer liver metastases: Evidence for involvement of MMP-7 activation in human cancer metastases. Clin Cancer Res. 8:144–148. 2002.PubMed/NCBI
24	Caruso R, Pallone F, Fina D, Gioia V, Peluso I, Caprioli F, Stolfi C, Perfetti A, Spagnoli LG, Palmieri G, et al: Protease-activated receptor-2 activation in gastric cancer cells promotes epidermal growth factor receptor trans-activation and proliferation. Am J Pathol. 169:268–278. 2006. View Article : Google Scholar : PubMed/NCBI
25	Darmoul D, Gratio V, Devaud H and Laburthe M: Protease-activated receptor 2 in colon cancer: Trypsin-induced MAPK phosphorylation and cell proliferation are mediated by epidermal growth factor receptor transactivation. J Biol Chem. 279:20927–20934. 2004. View Article : Google Scholar : PubMed/NCBI