Role of peptidylarginine deiminase type 4 in gastric cancer

Xin,Jiang; Song,Xiuqi

doi:10.3892/etm.2016.3798

Role of peptidylarginine deiminase type 4 in gastric cancer

Authors:
- Jiang Xin
- Xiuqi Song
View Affiliations

Affiliations: Qingdao University Medical College, Qingdao, Shandong 266003, P.R. China
Published online on: October 11, 2016 https://doi.org/10.3892/etm.2016.3798
Pages: 3155-3160
Copyright: © Xin 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

Peptidylarginine deiminase type 4 (PADI4) post-translationally converts peptidylarginine to citrulline, appearing to be overexpressed in numerous carcinomas. The current study aimed to investigate the expression of PADI4 in gastric cancer tissues and its effect on the biological activities of SGC‑7901 and AGS tumor cell lines. The expression of PADI4 was determined in gastric cancer and normal gastric mucosa tissues using western blot analysis and reverse transcription‑quantitative polymerase chain reaction. Gastric cancer cell lines were divided into the following groups: Mock group (subjected to transfection reagent); negative group [subjected to small interfering RNA (siRNA) transfection]; PADI4 siRNA group (subjected to PADI4 siRNA transfection); 5‑fluorouracil (5‑Fu) group (subjected to 5‑Fu); and 5‑Fu + siRNA transfection group (subjected to 5‑Fu and PADI4 siRNA transfection). The effects of silencing PADI4 with the above measures on the proliferation and invasion of SGC‑7901 and AGS cells were determined by MTT and Transwell chamber assays. In addition, propidium iodide staining was performed to detect the effects of PADI4 on the cell cycle. A significant increase in the expression of PADI4 mRNA in gastric cancer tissue compared with normal mucosa tissue was identified (P<0.05). The proliferation and invasion of SGC‑7901 and AGS cells were significantly decreased in the PADI4 siRNA group. Furthermore, flow cytometry DNA analysis revealed that silencing PADI4 resulted in significant S phase arrest and marked decrease of cells in the G2/M phase. PADI4 siRNA coupled with 5‑Fu significantly enhanced its inhibitory effect on the proliferation of gastric cancer cells. In conclusion, PADI4 demonstrated high expression in gastric cancer and served an important role in the biological activities of gastric cancer cells involving cell proliferation, invasion and cell cycle. As a result, PADI4 may be a valid cancer susceptibility gene and potential target for cancer therapy.

View Figures

View References

1	Roder DM: The epidemiology of gastric cancer. Gastric Cancer. 5(Suppl 1): 5–11. 2002. View Article : Google Scholar : PubMed/NCBI
2	Parkin DM, Bray FI and Devesa SS: Cancer burden in the year 2000. The global picture. Eur J Cancer. 37(Suppl 1): S4–S66. 2001. View Article : Google Scholar
3	Tahara E: Molecular aspects of invasion and metastasis of stomach cancer. Verh Dtsch Ges Pathol. 84:43–49. 2000.PubMed/NCBI
4	Sud R, Wells D, Talbot IC and Delhanty JD: Genetic alterations in gastric cancers from British patients. Cancer Genet Cytogenet. 126:111–119. 2001. View Article : Google Scholar : PubMed/NCBI
5	Fiocca R, Luinetti O, Villani L, Mastracci L, Quilici P, Grillo F and Ranzani GN: Molecular mechanisms involved in the pathogenesis of gastric carcinoma: Interactions between genetic alterations, cellular phenotype and cancer histotype. Hepatogastroenterology. 48:1523–1530. 2001.PubMed/NCBI
6	El-Rifai W and Powell SM: Molecular biology of gastric cancer. Semin Radiat Oncol. 12:128–140. 2002. View Article : Google Scholar : PubMed/NCBI
7	Wang L, Wei D, Huang S, Peng Z, Le X, Wu TT, Yao J, Ajani J and Xie K: Transcription factor Sp1 expression is a significant predictor of survival in human gastric cancer. Clin Cancer Res. 9:6371–6380. 2003.PubMed/NCBI
8	Kurayoshi M, Oue N, Yamamoto H, Kishida M, Inoue A, Asahara T, Yasui W and Kikuchi A: Expression of Wnt-5a is correlated with aggressiveness of gastric cancer by stimulating cell migration and invasion. Cancer Res. 66:10439–10448. 2006. View Article : Google Scholar : PubMed/NCBI
9	Wei D, Gong W, Kanai M, Schlunk C, Wang L, Yao JC, Wu TT, Huang S and Xie K: Drastic down-regulation of Krüppel-like factor 4 expression is critical in human gastric cancer development and progression. Cancer Res. 65:2746–2754. 2005. View Article : Google Scholar : PubMed/NCBI
10	Liu YL, Chiang YH, Liu GY and Hung HC: Functional role of dimerization of human peptidylarginine deiminase 4 (PAD4). PLoS One. 6:e213142011. View Article : Google Scholar : PubMed/NCBI
11	Chang X and Han J: Expression of peptidylarginine deiminase type 4 (PAD4) in various tumors. Mol Carcinog. 45:183–196. 2006. View Article : Google Scholar : PubMed/NCBI
12	Chang X, Han J, Pang L, Zhao Y, Yang Y and Shen Z: Increased PADI4 expression in blood and tissues of patients with malignant tumors. BMC Cancer. 9:402009. View Article : Google Scholar : PubMed/NCBI
13	Wang L, Chang X, Yuan G, Zhao Y and Wang P: Expression of peptidylarginine deiminase type 4 in ovarian tumors. Int J Biol Sciences. 6:454–464. 2010. View Article : Google Scholar
14	Ordóñez A, Yélamos J, Pedersen S, Miñano A, Conesa-Zamora P, Kristensen SR, Stender MT, Thorlacius-Ussing O, Martínez-Martínez I, Vicente V and Corral J: Increased levels of citrullinated antithrombin in plasma of patients with rheumatoid arthritis and colorectal adenocarcinoma determined by a newly developed ELISA using a specific monoclonal antibody. Thromb Haemost. 104:1143–1149. 2010. View Article : Google Scholar : PubMed/NCBI
15	Simms D, Cizdziel PE and Chomczynski P: TRIzol: A new reagent for optimal single-step isolation of RNA. Focus. 15:532–535. 1993.
16	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
17	Tiffen JC, Bailey CG, Ng C, Rasko JE and Holst J: Luciferase expression and bioluminescence does not affect tumor cell growth in vitro or in vivo. Mol Cancer. 9:2992010. View Article : Google Scholar : PubMed/NCBI
18	Haase SB and Reed SI: Improved flow cytometric analysis of the budding yeast cell cycle. Cell Cycle. 1:132–136. 2002. View Article : Google Scholar : PubMed/NCBI
19	Katz M, Amit I, Citri A, Shay T, Carvalho S, Lavi S, Milanezi F, Lyass L, Amariglio N, Jacob-Hirsch J, et al: A reciprocal tensin-3-cten switch mediates EGF-driven mammary cell migration. Nat Cell Biol. 9:961–969. 2007. View Article : Google Scholar : PubMed/NCBI
20	Wang Y, Wysocka J, Sayegh J, Lee YH, Perlin JR, Leonelli L, Sonbuchner LS, McDonald CH, Cook RG, Dou Y, et al: Human PAD4 regulates histone arginine methylation levels via demethylimination. Science. 306:279–283. 2004. View Article : Google Scholar : PubMed/NCBI
21	Klose RJ and Zhang Y: Regulation of histone methylation by demethylimination and demethylation. Nat Rev Mol Cell Biol. 8:307–318. 2007. View Article : Google Scholar : PubMed/NCBI
22	Cuthbert GL, Daujat S, Snowden AW, Erdjument-Bromage H, Hagiwara T, Yamada M, Schneider R, Gregory PD, Tempst P, Bannister AJ and Kouzarides T: Histone deimination antagonizes arginine methylation. Cell. 118:545–553. 2004. View Article : Google Scholar : PubMed/NCBI
23	Suzuki A, Yamada R, Chang X, Tokuhiro S, Sawada T, Suzuki M, Nagasaki M, Nakayama-Hamada M, Kawaida R, Ono M, et al: Functional haplotypes of PADI4, encoding citrullinating enzyme peptidylarginine deiminase 4, are associated with rheumatoid arthritis. Nat Genet. 34:395–402. 2003. View Article : Google Scholar : PubMed/NCBI
24	Chang X, Zhao Y, Sun S, Zhang Y and Zhu Y: The expression of PADI4 in synovium of rheumatoid arthritis. Rheumatol Int. 29:1411–1416. 2009. View Article : Google Scholar : PubMed/NCBI
25	Iwamoto T, Ikari K, Nakamura T, Kuwahara M, Toyama Y, Tomatsu T, Momohara S and Kamatani N: Association between PADI4 and rheumatoid arthritis: A meta-analysis. Rheumatology (Oxford). 45:804–807. 2006. View Article : Google Scholar : PubMed/NCBI
26	Lv Y, Xia Y, Wang Y and Cai C: Expression of PADI4 in hepatocellular carcinoma. Chinese-German J Clin Oncol. 8:453–455. 2009. View Article : Google Scholar
27	Ulivi P, Mercatali L, Casoni GL, Scarpi E, Bucchi L, Silvestrini R, Sanna S, Monteverde M, Amadori D, Poletti V and Zoli W: Multiple marker detection in peripheral blood for NSCLC diagnosis. PLoS One. 8:e574012013. View Article : Google Scholar : PubMed/NCBI
28	Kessenbrock K, Plaks V and Werb Z: Matrix metalloproteinases: Regulators of the tumor microenvironment. Cell. 141:52–67. 2010. View Article : Google Scholar : PubMed/NCBI
29	Bauvois B: New facets of matrix metalloproteinases MMP-2 and MMP-9 as cell surface transducers: Outside-in signaling and relationship to tumor progression. Biochim Biophys Acta. 1825:29–36. 2012.PubMed/NCBI
30	Roy R, Yang J and Moses MA: Matrix metalloproteinases as novel biomarkers and potential therapeutic targets in human cancer. J Clin Oncol. 27:5287–5297. 2009. View Article : Google Scholar : PubMed/NCBI
31	Turpeenniemi-Hujanen T: Gelatinases (MMP-2 and −9) and their natural inhibitors as prognostic indicators in solid cancers. Biochimie. 87:287–297. 2005. View Article : Google Scholar : PubMed/NCBI
32	Wong TS, Kwong DL, Sham JS, Wei WI, Kwong YL and Yuen AP: Clinicopathologic significance of plasma matrix metalloproteinase-2 and −9 levels in patients with undifferentiated nasopharyngeal carcinoma. Eur J Surg Oncol. 30:560–564. 2004. View Article : Google Scholar : PubMed/NCBI
33	Matsumura S, Oue N, Nakayama H, Kitadai Y, Yoshida K, Yamaguchi Y, Imai K, Nakachi K, Matsusaki K, Chayama K and Yasui W: A single nucleotide polymorphism in the MMP-9 promoter affects tumor progression and invasive phenotype of gastric cancer. J Cancer Res Clin Oncol. 131:19–25. 2005. View Article : Google Scholar : PubMed/NCBI
34	Liu GY, Liao YF, Chang WH, Liu CC, Hsieh MC, Hsu PC, Tsay GJ and Hung HC: Overexpression of peptidylarginine deiminase IV features in apoptosis of haematopoietic cells. Apoptosis. 11:183–196. 2006. View Article : Google Scholar : PubMed/NCBI
35	Li P, Yao H, Zhang Z, Li M, Luo Y, Thompson PR, Gilmour DS and Wang Y: Regulation of p53 target gene expression by peptidylarginine deiminase 4. Mol Cell Biol. 28:4745–4758. 2008. View Article : Google Scholar : PubMed/NCBI
36	Yao H, Li P, Venters BJ, Zheng S, Thompson PR, Pugh BF and Wang Y: Histone Arg modifications and p53 regulate the expression of OKL38, a mediator of apoptosis. J Biol Chem. 283:20060–20068. 2008. View Article : Google Scholar : PubMed/NCBI