Expression and regulation of nonsteroidal anti-inflammatory drug–activated gene (NAG-1) in human and mouse tissue

doi:10.1053/gast.2002.32972

Gastroenterology

Volume 122, Issue 5, May 2002, Pages 1388-1398

https://doi.org/10.1053/gast.2002.32972 Get rights and content

Abstract

Background & Aims: Nonsteroidal anti-inflammatory drugs (NSAIDs) induce NSAID-activated gene 1 (NAG-1), which has proapoptotic and antitumorigenic activities. However, NAG-1 expression and its relationship with apoptosis in human and mouse intestinal tract have not been determined. Methods: NAG-1 expression in human and mouse tissue was determined by immunohistochemistry, and apoptosis was estimated by in situ apoptosis detection. Apoptosis in NAG-1 overexpressing HCT-116 cells was examined with flow cytometry after cell sorting by green fluorescence protein. NAG-1 regulation in mouse cells was examined by Northern blot analysis, comparing sulindac-treated and nontreated mice. Results: Apoptosis was higher in NAG-1 overexpressing cells compared with controls. Human NAG-1 protein was localized to the colonic surface epithelium where cells undergo apoptosis, and higher expression was observed in the normal surface epithelium than in most of the tumors. This localization and lower expression in tumors was similar to that in the Min mouse, in which NSAIDs were also shown to regulate the expression of NAG-1 in mouse cells. Sulindac treatment of mice increased the NAG-1 expression in the colon and liver. Conclusions: Based on these results, we propose that NAG-1 acts as a mediator of apoptosis in intestinal cells and may contribute to cancer chemoprevention by NSAIDs.

GASTROENTEROLOGY 2002;122:1388-1398

Section snippets

Reagents and cell lines

pEGFP-N2 and anti-green fluorescence protein (GFP) monoclonal antibody were purchased from Clontech Laboratories, Inc. (Palo Alto, CA). Two polyclonal rabbit anti-human NAG-1 antibodies were used. One described previously was used for human cells and tissue.⁷ The second was a generous gift from Dr. Vishwas Paraklar (Pfizer Inc., Groton, CT) and was used for mouse studies. Monoclonal anti–proliferating cell nuclear antigen (PCNA) antibody was obtained from Santa Cruz Biotechnology, Inc. (Santa

Localization of NAG-1 protein in human colorectal tumor and normal tissue

NAG-1 protein expression and localization in human colorectal tumor and normal tissue was examined by immunohistochemistry using 25 matched pairs of tissues. Thirteen pairs were from males and 12 were from females. The degree of differentiation, tumor localization, histology, and Duke's stage are described in Table 1.The tumors included 2 villotubular adenomas and 23 adenocarcinomas (2 of which were mucinous) ranging from well to poorly differentiated. In normal colon mucosa, NAG-1 protein

Discussion

NAG-1 is a newly identified member of the TGF-β superfamily that only shares 25% sequence identity to other family members.⁷ The biological activity of NAG-1 is not fully characterized, but it seems to have various functions. For example, NAG-1 induces cartilage and bone formation and suppresses inflammation by inhibiting macrophage activation.12, 26 Other reports provide some clues of antitumorigenic and proapoptotic activities.7, 10, 11 In this study, we confirmed the apoptotic ability of

Acknowledgements

The authors thank Dr. Julie Foley for helpful suggestions on the immunohistochemical analysis, Evangeline Raynolds of the University of North Carolina for assistance in obtaining the human tissue samples, Dr. Carl Bortner for helping with the flow cytometry analysis, Dr. Joseph Haseman and Dr. Jung Wan Koo for the analysis of the data, and Dr. Hiroo Kawajiri, Dr. Jennifer Nixon, Dr. Hideto Kameda, and Allison Call for their assistance and comments on the manuscript.

References (37)

SD Markowitz et al.
Tumor suppressor activity of the TGF-beta pathway in human cancers
Cytokine Growth Factor Rev
(1996)
LN Lawton et al.
Identification of a novel member of the TGF-beta superfamily highly expressed in human placenta
Gene
(1997)
PX Li et al.
Placental transforming growth factor-beta is a downstream mediator of the growth arrest and apoptotic response of tumor cells to DNA damage and p53 overexpression
J Biol Chem
(2000)
R Hromas et al.
PLAB, a novel placental bone morphogenetic protein
Biochim Biophys Acta
(1997)
M Bottner et al.
Characterization of the rat, mouse, and human genes of growth/differentiation factor-15/macrophage inhibiting cytokine-1 (GDF- 15/MIC-1)
Gene
(1999)
SJ Baek et al.
Molecular cloning and characterization of human NSAID activated gene, NAG-1, promoter: basal transcription is mediated by Sp1 and Sp3
J Biol Chem
(2001)
SF Moss et al.
Inward growth of colonic adenomatous polyps
Gastroenterology
(1996)
OC Trifan et al.
Overexpression of cyclooxygenase-2 induces cell cycle arrest. Evidence for a prostaglandin-independent mechanism
J Biol Chem
(1999)
NN Mahmoud et al.
Aspirin prevents tumors in a murine model of familial adenomatous polyposis
Surgery
(1998)
VM Paralkar et al.
Cloning and characterization of a novel member of the transforming growth factor-beta/bone morphogenetic protein family
J Biol Chem
(1998)

BP Cormack et al.

FACS-optimized mutants of the green fluorescent protein (GFP)

Gene

(1996)

K Kannan et al.

Profile of gene expression regulated by induced p53: connection to the TGF-beta family

FEBS Lett

(2000)

I Schwarte-Waldhoff et al.

DPC4/SMAD4 mediated tumor suppression of colon carcinoma cells is associated with reduced urokinase expression

Oncogene

(1999)

SK Boolbol et al.

Cyclooxygenase-2 overexpression and tumor formation are blocked by sulindac in a murine model of familial adenomatous polyposis

Cancer Res

(1996)

CH Chiu et al.

Sulindac causes rapid regression of preexisting tumors in Min/+ mice independent of prostaglandin biosynthesis

Cancer Res

(1997)

FM Giardiello et al.

Treatment of colonic and rectal adenomas with sulindac in familial adenomatous polyposis

N Engl J Med

(1993)

E Hawk et al.

Chemoprevention in hereditary colorectal cancer syndromes

Cancer

(1999)

SJ Baek et al.

Cyclooxygenase inhibitors regulate the expression of a TGF-β superfamily member that has pro-apoptotic and anti-tumorigenic activities

Mol Phamacol

(2001)

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^☆: Address requests for reprints to: Thomas Eling, M.D., Laboratory of Molecular Carcinogenesis, NIEHS, 111 TW Alexander Drive, Research Triangle Park, North Carolina 27709. e-mail: [email protected]; fax: (919) 541-0146

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Basic ResearchExpression and regulation of nonsteroidal anti-inflammatory drug–activated gene (NAG-1) in human and mouse tissue☆

Abstract

Section snippets

Reagents and cell lines

Localization of NAG-1 protein in human colorectal tumor and normal tissue

Discussion

Acknowledgements

Cytokine Growth Factor Rev

Gene

J Biol Chem

Biochim Biophys Acta

Gene

J Biol Chem

Gastroenterology

J Biol Chem

Surgery

J Biol Chem

Gene

FEBS Lett

DPC4/SMAD4 mediated tumor suppression of colon carcinoma cells is associated with reduced urokinase expression

Oncogene

Cyclooxygenase-2 overexpression and tumor formation are blocked by sulindac in a murine model of familial adenomatous polyposis

Cancer Res

Sulindac causes rapid regression of preexisting tumors in Min/+ mice independent of prostaglandin biosynthesis

Cancer Res

Treatment of colonic and rectal adenomas with sulindac in familial adenomatous polyposis

N Engl J Med

Chemoprevention in hereditary colorectal cancer syndromes

Cancer

Cyclooxygenase inhibitors regulate the expression of a TGF-β superfamily member that has pro-apoptotic and anti-tumorigenic activities

Mol Phamacol

Basic Research
Expression and regulation of nonsteroidal anti-inflammatory drug–activated gene (NAG-1) in human and mouse tissue☆