Loss of trefoil factor 1 is regulated by DNA methylation and is an independent predictive factor for poor survival in advanced gastric cancer

Tanaka,Tomokazu; Nakamura,Jun; Kitajima,Yoshihiko; Kai,Keita; Miyake,Shuusuke; Hiraki,Masatsugu; Ide,Takao; Koga,Yasuo; Noshiro,Hirokazu

doi:10.3892/ijo.2013.1759

Loss of trefoil factor 1 is regulated by DNA methylation and is an independent predictive factor for poor survival in advanced gastric cancer

Authors:
- Tomokazu Tanaka
- Jun Nakamura
- Yoshihiko Kitajima
- Keita Kai
- Shuusuke Miyake
- Masatsugu Hiraki
- Takao Ide
- Yasuo Koga
- Hirokazu Noshiro
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Affiliations: Department of Surgery, Saga University Faculty of Medicine, Saga 849-8501, Japan, Department of Pathology and Microbiology, Saga University Faculty of Medicine, Saga 849-8501, Japan
Published online on: January 3, 2013 https://doi.org/10.3892/ijo.2013.1759
Pages: 894-902

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Abstract

Trefoil factor 1 (TFF1) is considered to be a tumor suppressor gene in gastric cancer. However, the role of TFF1 expression and its regulation in gastric cancer patients remain unclear. The aims of this study were to clarify the clinical significance of TFF1 and to determine its regulatory mechanisms. We assessed the immunohistochemical expression of TFF1 in 182 gastric cancer patients and examined whether or not TFF1 is associated with the clinicopathological factors and patient survival. In vitro study using TFF1 knockdown gastric cancer cells evaluated the role of TFF1 in cancer invasion. Bisulfite sequencing was performed to assess DNA methylation of TFF1 in cells and resected tissues. Patients with low expression of TFF1 showed a significantly deeper invasion of the tumor than those with high expression (p=0.037). Low expression of TFF1 was also associated with a poor survival (p=0.029) in 108 patients who were treated by surgery alone. Both TFF1 expression and lymph node metastasis are independent predictive factors for disease‑specific survival in a multivariate analysis. In an in vitro study, invasive power of the cells was significantly increased in the TFF1‑deficient cells compared with the control cells. Bisulfate sequencing showed that TFF1 expression is strongly dependent on DNA methylation in both gastric cancer cells and tissues. Interestingly, methylation status of two specific CpG sites, which are located close to a TATA box and hypoxia response element (HRE), determined the TFF1 expression in the resected tissues. TFF1 expression is silenced by DNA methylation and is associated with tumor invasion and a poor survival in gastric cancer patients. The expression and̸or methylation status of TFF1 may, therefore, serve as a useful biomarker for predicting survival in patients with advanced gastric cancer.

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1	Wong WM, Poulsom R and Wright NA: Trefoil peptides. Gut. 44:890–895. 1999. View Article : Google Scholar : PubMed/NCBI
2	Hoffmann W and Jagla W: Cell type specific expression of secretory TFF peptides: colocalization with mucins and synthesis in the brain. Int Rev Cytol. 213:147–181. 2002. View Article : Google Scholar : PubMed/NCBI
3	Taupin D and Podolsky DK: Trefoil factors: initiators of mucosal healing. Nat Rev Mol Cell Biol. 4:721–732. 2003. View Article : Google Scholar : PubMed/NCBI
4	Thim L: Trefoil peptides: from structure to function. Cell Mol Life Sci. 53:888–903. 1997. View Article : Google Scholar : PubMed/NCBI
5	Masiakowski P, Breathnach R, Bloch J, Gannon F, Krust A and Chambon P: Cloning of cDNA sequences of hormone-regulated genes from the MCF-7 human breast cancer cell line. Nucleic Acids Res. 10:7895–7903. 1982. View Article : Google Scholar : PubMed/NCBI
6	Rio MC, Bellocq JP, Daniel JY, et al: Breast cancer-associated pS2 protein: synthesis and secretion by normal stomach mucosa. Science. 241:705–708. 1988. View Article : Google Scholar : PubMed/NCBI
7	Tomasetto C, Masson R, Linares JL, Wendling C, Lefebvre O, Chenard MP and Rio MC: pS2/TFF1 interacts directly with the VWFC cysteine-rich domains of mucins. Gastroenterology. 118:70–80. 2000. View Article : Google Scholar : PubMed/NCBI
8	Hoffmann W, Jagla W and Wiede A: Molecular medicine of TFF-peptides: from gut to brain. Histol Histopathol. 16:319–334. 2001.PubMed/NCBI
9	Lefebvre O, Chenard MP, Masson R, et al: Gastric mucosa abnormalities and tumorigenesis in mice lacking the pS2 trefoil protein. Science. 274:259–262. 1996. View Article : Google Scholar : PubMed/NCBI
10	Colombel M, Dante R, Bouvier R, Ribieras S, Pangaud C, Marechal JM and Lasne Y: Differential RNA expression of the pS2 gene in the human benign and malignant prostatic tissue. J Urol. 162:927–930. 1999. View Article : Google Scholar : PubMed/NCBI
11	Buache E, Etique N, Alpy F, et al: Deficiency in trefoil factor 1 (TFF1) increases tumorigenicity of human breast cancer cells and mammary tumor development in TFF1-knockout mice. Oncogene. 30:3261–3273. 2011. View Article : Google Scholar : PubMed/NCBI
12	Arumugam T, Brandt W, Ramachandran V, et al: Trefoil factor 1 stimulates both pancreatic cancer and stellate cells and increases metastasis. Pancreas. 40:815–822. 2011. View Article : Google Scholar : PubMed/NCBI
13	McChesney PA, Aiyar SE, Lee OJ, Zaika A, Moskaluk C, Li R and El-Rifai W: Cofactor of BRCA1: a novel transcription factor regulator in upper gastrointestinal adenocarcinomas. Cancer Res. 66:1346–1353. 2006. View Article : Google Scholar
14	Milne AN, Carvalho R, Morsink FM, Musler AR, de Leng WW, Ristimäki A and Offerhaus GJ: Early-onset gastric cancers have a different molecular expression profile than conventional gastric cancers. Mod Pathol. 19:564–572. 2006. View Article : Google Scholar : PubMed/NCBI
15	Gött P, Beck S, Machado JC, Carneiro F, Schmitt H and Blin N: Human trefoil peptides: genomic structure in 21q22.3 and coordinated expression. Eur J Hum Genet. 4:308–315. 1996.PubMed/NCBI
16	Beck S, Sommer P, Blin N and Gött P: 5′-flanking motifs control cell-specific expression of trefoil factor genes (TFF). Int J Mol Med. 2:353–361. 1998.
17	Al-azzeh ED, Fegert P, Blin N and Gött P: Transcription factor GATA-6 activates expression of gastroprotective trefoil genes TFF1 and TFF2. Biochim Biophys Acta. 1490:324–332. 2000. View Article : Google Scholar : PubMed/NCBI
18	Beck S, Sommer P, dos Santos Silva E, Blin N and Gött P: Hepatocyte nuclear factor 3 (winged helix domain) activates trefoil factor gene TFF1 through a binding motif adjacent to the TATAA box. DNA Cell Biol. 18:157–164. 1999. View Article : Google Scholar
19	Hernández C, Santamatilde E, McCreath KJ, et al: Induction of trefoil factor (TFF) 1, TFF2 and TFF3 by hypoxia is mediated by hypoxia inducible factor-1: implications for gastric mucosal healing. Br J Pharmacol. 156:262–272. 2009.PubMed/NCBI
20	Japanese Gastric Cancer Association: Japanese classification of gastric carcinoma: 3rd English ed. Gastric Cancer. 14:101–112. 2011. View Article : Google Scholar
21	Yanagihara K, Ito A, Toge T and Numoto M: Antiproliferative effects of isoflavones on human cancer cell lines established from the gastrointestinal tract. Cancer Res. 53:5815–5821. 1993.PubMed/NCBI
22	Yanagihara K, Tanaka H, Takigahira M, et al: Establishment of two cell lines from human gastric scirrhous carcinoma that possess the potential to metastasize spontaneously in nude mice. Cancer Sci. 95:575–582. 2004. View Article : Google Scholar : PubMed/NCBI
23	Nakamura J, Kitajima Y, Kai K, et al: Expression of hypoxic marker CA IX is regulated by site-specific DNA methylation and is associated with the histology of gastric cancer. Am J Pathol. 178:515–524. 2011. View Article : Google Scholar : PubMed/NCBI
24	Babyatsky MW, deBeaumont M, Thim L and Podolsky DK: Oral trefoil peptides protect against ethanol- and indomethacin-induced gastric injury in rats. Gastroenterology. 110:489–497. 1996. View Article : Google Scholar : PubMed/NCBI
25	Playford RJ, Marchbank T, Goodlad RA, Chinery RA, Poulsom R and Hanby AM: Transgenic mice that overexpress the human trefoil peptide pS2 have an increased resistance to intestinal damage. Proc Natl Acad Sci USA. 93:2137–2142. 1996. View Article : Google Scholar : PubMed/NCBI
26	Peterson DE, Barker NP, Akhmadullina LI, et al: Phase II, randomized, double-blind, placebo-controlled study of recombinant human intestinal trefoil factor oral spray for prevention of oral mucositis in patients with colorectal cancer who are receiving fluorouracil-based chemotherapy. J Clin Oncol. 27:4333–4338. 2009. View Article : Google Scholar
27	Park WS, Oh RR, Park JY, et al: Mapping of a new target region of allelic loss at 21q22 in primary gastric cancers. Cancer Lett. 159:15–21. 2000. View Article : Google Scholar : PubMed/NCBI
28	Park WS, Oh RR, Park JY, et al: Somatic mutations of the trefoil factor family 1 gene in gastric cancer. Gastroenterology. 119:691–698. 2000. View Article : Google Scholar : PubMed/NCBI
29	Carvalho R, Kayademir T, Soares P, et al: Loss of heterozygosity and promoter methylation, but not mutation, may underlie loss of TFF1 in gastric carcinoma. Lab Invest. 82:1319–1326. 2002. View Article : Google Scholar : PubMed/NCBI
30	Beckler AD, Roche JK, Harper JC, et al: Decreased abundance of trefoil factor 1 transcript in the majority of gastric carcinomas. Cancer. 98:2184–2191. 2003. View Article : Google Scholar : PubMed/NCBI
31	Yio X, Diamond M, Zhang JY, Weinstein H, Wang LH, Werther L and Itzkowitz S: Trefoil factor family-1 mutations enhance gastric cancer cell invasion through distinct signaling pathways. Gastroenterology. 130:1696–1706. 2006. View Article : Google Scholar : PubMed/NCBI
32	Lifton RP, Goldberg ML, Karp RW and Hogness DS: The organization of the histone genes in Drosophila melanogaster: functional and evolutionary implications. Cold Spring Harb Symp Quant Biol. 42:1047–1051. 1978.
33	Semenza GL, Jiang BH, Leung SW, Passantino R, Concordet JP, Maire P and Giallongo A: Hypoxia response elements in the aldolase A, enolase 1, and lactate dehydrogenase A gene promoters contain essential binding sites for hypoxia-inducible factor 1. J Biol Chem. 271:32529–32537. 1996. View Article : Google Scholar