Helicobacter pylori infection promotes methylation and silencing of trefoil factor 2, leading to gastric tumor development in mice and humans.
OBJECTIVE
Trefoil factors (TFFs) regulate mucosal repair and suppress tumor formation in the stomach. Tff1 deficiency results in gastric cancer, whereas Tff2 deficiency increases gastric inflammation. TFF2 expression is frequently lost in gastric neoplasms, but the nature of the silencing mechanism and associated impact on tumorigenesis have not been determined.


METHODS
We investigated the epigenetic silencing of TFF2 in gastric biopsy specimens from individuals with Helicobacter pylori-positive gastritis, intestinal metaplasia, gastric cancer, and disease-free controls. TFF2 function and methylation were manipulated in gastric cancer cell lines. The effects of Tff2 deficiency on tumor growth were investigated in the gp130(F/F) mouse model of gastric cancer.


RESULTS
In human tissue samples, DNA methylation at the TFF2 promoter began at the time of H pylori infection and increased throughout gastric tumor progression. TFF2 methylation levels were inversely correlated with TFF2 messenger RNA levels and could be used to discriminate between disease-free controls, H pylori-infected, and tumor tissues. Genome demethylation restored TFF2 expression in gastric cancer cell lines, so TFF2 silencing requires methylation. In Tff2-deficient gp130(F/F)/Tff2(-/-) mice, proliferation of mucosal cells and release of T helper cell type-1 (Th-1) 1 cytokines increased, whereas expression of gastric tumor suppressor genes and Th-2 cytokines were reduced, compared with gp130(F/F)controls. The fundus of gp130(F/F)/Tff2(-/-) mice displayed glandular atrophy and metaplasia, indicating accelerated preneoplasia. Experimental H pylori infection in wild-type mice reduced antral expression of Tff2 by increased promoter methylation.


CONCLUSIONS
TFF2 negatively regulates preneoplastic progression and subsequent tumor development in the stomach, a role that is subverted by promoter methylation during H pylori infection.
Peterson
Anthony J
AJ
Murdoch Children's Research Institute, Royal Children's Hospital, Flemington Road, Parkville, Victoria.
Menheniott
Trevelyan R
TR
O'Connor
Louise
L
Walduck
Anna K
AK
Fox
James G
JG
Kawakami
Kazuyuki
K
Minamoto
Toshinari
T
Ong
Eng Kok
EK
Wang
Timothy C
TC
Judd
Louise M
LM
Giraud
Andrew S
AS
D000818:Animals; D001284:Atrophy; D045744:Cell Line, Tumor; D050822:Cytokine Receptor gp130 / Q000235:genetics; D019175:DNA Methylation / Q000502:physiology; D057890:Epigenomics; D005756:Gastritis / Q000235:genetics / Q000382:microbiology / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology; D020868:Gene Silencing / Q000502:physiology; D016481:Helicobacter Infections / Q000235:genetics* / Q000473:pathology; D016480:Helicobacter pylori; D006801:Humans; D008679:Metaplasia; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D009077:Mucins / Q000235:genetics*; D009124:Muscle Proteins / Q000235:genetics*; D010455:Peptides / Q000235:genetics*; D015398:Signal Transduction / Q000502:physiology; D013274:Stomach Neoplasms / Q000235:genetics / Q000382:microbiology / Q000473:pathology; D018417:Th1 Cells / Q000502:physiology; D000071164:Trefoil Factor-2
C494200:Il6st protein, mouse; D009077:Mucins; D009124:Muscle Proteins; D010455:Peptides; C000604007:TFF2 protein, human; C481546:TFF2 protein, mouse; D000071164:Trefoil Factor-2; D050822:Cytokine Receptor gp130
Biochemistry. 2005 Jun 7;44(22):7967-75
15924415
Cell Mol Life Sci. 2005 Dec;62(24):2921-31
16374580
Nat Med. 2002 Oct;8(10):1089-97
12219085
Infect Immun. 2007 Jan;75(1):471-80
17101660
Gastroenterology. 2003 Aug;125(2):510-21
12891554
J Cell Biol. 2002 May 27;157(5):761-70
12034770
Gastroenterology. 1998 Apr;114(4):675-89
9516388
Gastroenterology. 1995 Aug;109(2):516-23
7615201
Annu Rev Physiol. 2001;63:119-39
11181951
Peptides. 1995;16(6):1001-5
8532580
Science. 1996 Oct 11;274(5285):259-62
8824193
Nat Rev Mol Cell Biol. 2003 Sep;4(9):721-32
14506475
Lab Invest. 2002 Oct;82(10):1319-26
12379766
Clin Cancer Res. 2008 Jul 1;14(13):4161-7
18593995
FEBS Lett. 1997 May 19;408(2):121-3
9187350
J Clin Invest. 1994 Jul;94(1):376-83
8040278
Cancer Cell. 2008 Nov 4;14(5):408-19
18977329
Cell Physiol Biochem. 2005;16(1-3):31-42
16121031
J Clin Invest. 2002 Jan;109(2):193-204
11805131
Cancer Res. 2003 May 15;63(10):2569-77
12750281
Lab Invest. 1999 Jun;79(6):639-46
10378506
Mol Cell Biol. 2003 Dec;23(23):8429-39
14612389
Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11017-21
1763017
J Gastroenterol Hepatol. 1998 Apr;13(4):363-70
9641298
Biochem Biophys Res Commun. 2002 Apr 26;293(1):366-74
12054609
Am J Physiol Gastrointest Liver Physiol. 2007 Jan;292(1):G1-5
16935852
Nucleic Acids Res. 2007;35(18):e119
17855397
J Exp Med. 1999 Dec 6;190(11):1595-604
10587350
Am J Physiol Gastrointest Liver Physiol. 2005 Feb;288(2):G362-75
15647607
Am J Physiol Gastrointest Liver Physiol. 2006 Apr;290(4):G793-804
16306133
J Pathol. 1999 Jul;188(3):312-7
10419601
Oncogene. 2005 Mar 31;24(14):2354-66
15735748
Gastroenterology. 1993 Jan;104(1):12-20
8419234
J Cell Biol. 1993 Jul;122(1):191-8
8314841
Cancer Res. 2002 Feb 1;62(3):696-702
11830522
Gastroenterology. 1992 Jun;102(6):2177-8
1587446
Am J Pathol. 2007 Nov;171(5):1520-8
17982128
Aliment Pharmacol Ther. 2008 Sep 15;28(6):782-8
19145733
Int J Dev Biol. 1998 Sep;42(6):783-9
9727834
J Pathol. 2007 Oct;213(2):140-51
17724739
Am J Pathol. 2003 Oct;163(4):1551-6
14507661
J Pathol. 2002 Aug;197(5):582-8
12210076
Gastroenterology. 2004 Jan;126(1):196-207
14699500
Scand J Gastroenterol. 2009;44(3):339-45
19031173
Gut. 2006 Dec;55(12):1717-24
16641130
Am J Physiol Gastrointest Liver Physiol. 2003 Aug;285(2):G332-43
12851218
Gastroenterology. 2006 Oct;131(4):1073-85
17030178
World J Gastroenterol. 2006 May 21;12(19):3119-22
16718800
Gastroenterology. 1995 Jan;108(1):108-16
7806031
Gut. 2004 Oct;53(10):1408-15
15361486