Cyclooxygenase 2 expression is increased in the stroma of colon carcinomas from IL–10−/− mice

doi:10.1016/S0016-5085(00)70216-2

Gastroenterology

Volume 118, Issue 2, February 2000, Pages 337-345

https://doi.org/10.1016/S0016-5085(00)70216-2 Get rights and content

Abstract

Background & Aims: The pathological and molecular changes associated with colitis-associated colorectal cancer and sporadic colorectal cancer are considered to be distinct. Therefore, we have used a mouse model of ulcerative colitis to determine if expression of the enzyme cyclooxygenase (COX)-2 is increased in colitis-associated tumors. Methods: Reverse-transcription polymerase chain reaction and Western analysis were used to determine if COX-2 expression is increased in these tumors; in situ hybridization and immunohistochemistry were used to determine the localization of COX-2. Results: Increased levels of COX-2 messenger RNA and protein were detected in interleukin (IL)-10 (−/−) tumors and in an inflamed region of the colon that contained no macroscopically detected tumors. This expression was localized to the inflammatory cells associated with ulcerated regions of the tumor by in situ hybridization and immunohistochemistry. Increased COX-2 expression was also associated with the areas of the tumor expressing α–smooth muscle actin, which is a molecular marker for subepithelial myofibroblasts. The association between COX-2 expression and subepithelial myofibroblasts was also noted in tumors derived from the multiple intestinal neoplasia mice (Min/+) and from carcinogen-induced tumors. Conclusions: These results indicate that COX-2 is expressed very early in the pathogenesis of colitis-associated tumors, and that the expression pattern is similar to that seen in tumors from azoxymethane-treated and Min/+ mice.

GASTROENTEROLOGY 2000;118:337-345

Section snippets

Mice

Breeding pairs of IL-10^−/− mice on a C57BL/10 background were purchased from Jackson Laboratory (Bar Harbor, ME) and maintained in specific pathogen–free conditions in microisolator cages. Offspring were weaned at 3 weeks and moved to conventional conditions. The mice were killed at ages 40–45 weeks. C57Bl/6-Min/+ mice (Jackson Laboratory) were maintained and genotyped as described previously.²³ Wild-type C57Bl/6J mice treated with the colon carcinogen AOM (Sigma Chemical Co., St. Louis, MO)

COX-2 messenger RNA and protein expression is increased in colon tumors derived from IL^−/− mice

Tumors from IL-10^−/− mice are localized primarily in the cecum and proximal colon. Therefore, initially, COX-2 expression was compared between RNA samples derived from the proximal and distal colon segments by means of RT-PCR. As shown in Figure 1A, COX-2 messenger RNA (mRNA) could be detected in both the proximal and distal colon; however, the levels were higher in 4 of the 4 (100%) proximal colon samples.

. COX-2 mRNA expression is increased in IL-10^−/− tumors. (A) Proximal (PC) and distal (DC)

Discussion

There are 3 major classifications of colorectal cancer: hereditary colorectal cancer, sporadic colorectal cancer (SCC), and colitis-associated colorectal cancer (CAC). Each has distinct pathological and molecular alterations involved in the progression to carcinoma formation. SCC usually arises from an adenomatous polyp, and the dysplasia often occurs before development of cancer. CAC arises from areas of flat dysplasia, and the dysplastic epithelium in UC may be present in relatively large

Acknowledgements

The authors thank Debi Schweder for help with the mice, Sandy Olson for the α–smooth muscle actin staining, and Dr. Hossam Kandil for all of his enthusiasm toward this project. The authors also thank Dr. Don Powell for helpful discussions concerning the localization of COX-2 in myofibroblasts.

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^☆: Supported by National Institutes of Health grants DK47297 and CA77839 (to R.N.D.), grant DK02401-01A (to G.W.V.), and VA Merit Award (R.N.D.). Dr. DuBois is the Mina C. Wallace Professor of Gastroenterology and Cancer Prevention.

^☆☆: Address requests for reprints to: Raymond N. DuBois, Division of Gastroenterology, Department of Medicine, Vanderbilt University School of Medicine, MCN C-2102, 1161 21st Avenue South, Nashville, Tennessee 37232-2279. e-mail: [email protected]; fax: (615) 343-6229.

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Alimentary TractCyclooxygenase 2 expression is increased in the stroma of colon carcinomas from IL–10−/− mice☆,☆☆

Abstract

Section snippets

Mice

COX-2 messenger RNA and protein expression is increased in colon tumors derived from IL−/− mice

Discussion

Acknowledgements

Gastroenterol Clin North Am

Cell

Cell

Gastroenterology

J Biol Chem

Gastroenterology

Gastroenterology

Cell

Hum Pathol

Gastroenterology

Exp Pathol

Crohn's disease and colorectal carcinoma: rectal cancer complicating longstanding active perianal disease

Am J Gastroenterol

Lower gastrointestinal malignancy in Crohn's disease

Gut

Adenomatous polyposis coli gene mutations in ulcerative colitis–associated dysplasias and cancers versus sporadic colon neoplasms

Cancer Res

Identification of a phorbol ester–repressible v-src–inducible gene

Proc Natl Acad Sci USA

Prostaglandin endoperoxide synthase: why two isoforms?

Am J Physiol

Expression of cyclooxygenase-1 and -2 in human colorectal cancer

Cancer Res

Alimentary Tract
Cyclooxygenase 2 expression is increased in the stroma of colon carcinomas from IL–10^−/− mice☆,☆☆

COX-2 messenger RNA and protein expression is increased in colon tumors derived from IL^−/− mice