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Published in: BMC Gastroenterology 1/2014

Open Access 01-12-2014 | Research article

Over-expression of COX-2 mRNA in colorectal cancer

Authors: Hennie MJ Roelofs, Rene HM te Morsche, Bjorn WH van Heumen, Fokko M Nagengast, Wilbert HM Peters

Published in: BMC Gastroenterology | Issue 1/2014

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Abstract

Background

Cyclooxygenase-2 (COX-2, PTGS2) is an enzyme involved in the synthesis of prostaglandins and thromboxanes, which are regulators of biologic processes such as inflammation, cell proliferation and angiogenesis. COX-2 over-expression was reported in many (pre) malignant tissues, but data strongly vary and seem to depend on the methodology used.

Methods

Normal colorectal mucosa and paired cancerous tissue from 60 patients with colorectal cancer was investigated for the levels of COX-2 mRNA by real-time quantitative Polymerase Chain Reaction (qPCR). COX-2 levels were expressed relative to either: tissue weight or levels of the housekeeping genes beta-2 microglobulin (B2M) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH).

Results

COX-2 mRNA levels, normalized with respect to tissue weight or mRNA levels of the housekeeping genes B2M or GAPDH, were over-expressed in 80%, 70% and 40% of the colorectal tumor tissues, as compared to the paired adjacent normal colorectal mucosa samples, respectively. Highest mRNA COX-2 ratios tumor/normal were measured when expressed per mg tissue (mean ratio 21.6). When normalized with respect to the housekeeping genes B2M or GAPDH, mean tumor/normal ratios were 16.1 and 7.5, respectively.

Conclusion

Expression of COX-2 mRNA levels per mg tissue is most simple in comparison to normalization with respect to the housekeeping genes B2M or GAPDH. Levels of COX-2 mRNA are found over-expressed in almost 80% of the colorectal tumors, compared to paired adjacent normal colorectal mucosa, suggesting a role of COX-2 as a potential biomarker for cancer risk, whereas inhibitors of COX-2 could be of value in chemoprevention of colon cancer.
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Literature
2.
go back to reference Chandrasekharan NV, Dai H, Roos KL, Evanson NK, Tomsik J, Elton TS, Simmons DL: COX-3, a cyclooxygenase-1 variant inhibited by acetaminophen and other analgesic/antipyretic drugs: cloning, structure and expression. Proc Natl Acad Sci U S A. 2002, 99: 13926-13931. 10.1073/pnas.162468699.CrossRefPubMedPubMedCentral Chandrasekharan NV, Dai H, Roos KL, Evanson NK, Tomsik J, Elton TS, Simmons DL: COX-3, a cyclooxygenase-1 variant inhibited by acetaminophen and other analgesic/antipyretic drugs: cloning, structure and expression. Proc Natl Acad Sci U S A. 2002, 99: 13926-13931. 10.1073/pnas.162468699.CrossRefPubMedPubMedCentral
3.
go back to reference Sano H, Kawahito Y, Wilder RL, Hashiramoto A, Mukai S, Asai K, Kimura S, Kato H, Kondo M, Hla T: Expression of cyclooxygenase-1 and −2 in human colorectal cancer. Cancer Res. 1995, 55: 3785-3789.PubMed Sano H, Kawahito Y, Wilder RL, Hashiramoto A, Mukai S, Asai K, Kimura S, Kato H, Kondo M, Hla T: Expression of cyclooxygenase-1 and −2 in human colorectal cancer. Cancer Res. 1995, 55: 3785-3789.PubMed
4.
go back to reference Eberhart CE, Coffey RJ, Radhika A, Giardiello FM, Ferrenbach S, DuBois RN: Up-regulation of cyclooxygenase 2 gene expression in human colorectal adenomas and adenocarcinomas. Gastroenterology. 1994, 107: 1183-1188.CrossRefPubMed Eberhart CE, Coffey RJ, Radhika A, Giardiello FM, Ferrenbach S, DuBois RN: Up-regulation of cyclooxygenase 2 gene expression in human colorectal adenomas and adenocarcinomas. Gastroenterology. 1994, 107: 1183-1188.CrossRefPubMed
5.
go back to reference Fujimura T, Ohta T, Oyama K, Miyashita T, Miwa K: Role of cyclooxygenase-2 in the carcinogenesis of gastrointestinal tract cancers: a review and report of personal experience. World J Gastroenterology. 2006, 12: 1336-1345.CrossRef Fujimura T, Ohta T, Oyama K, Miyashita T, Miwa K: Role of cyclooxygenase-2 in the carcinogenesis of gastrointestinal tract cancers: a review and report of personal experience. World J Gastroenterology. 2006, 12: 1336-1345.CrossRef
6.
go back to reference Brown JR, DuBois RN: COX-2: a molecular target for colorectal cancer prevention. J Clin Oncol. 2005, 23: 2840-2855.CrossRefPubMed Brown JR, DuBois RN: COX-2: a molecular target for colorectal cancer prevention. J Clin Oncol. 2005, 23: 2840-2855.CrossRefPubMed
7.
go back to reference Mehta S, Boddy A, Johnson T, Rhodes M: Systematic review: cyclo-oxygenase-2 in human oesophageal adenocarcinogenesis. Aliment Pharmacol Ther. 2006, 24: 1321-1331. 10.1111/j.1365-2036.2006.03119.x.CrossRefPubMed Mehta S, Boddy A, Johnson T, Rhodes M: Systematic review: cyclo-oxygenase-2 in human oesophageal adenocarcinogenesis. Aliment Pharmacol Ther. 2006, 24: 1321-1331. 10.1111/j.1365-2036.2006.03119.x.CrossRefPubMed
8.
go back to reference Liu X, Li P, Zhang ST, You H, Jia JD, Yu ZL: COX-2 mRNA expression in esophageal squamous cell carcinoma (ESCC) and effect by NSAID. Dis Esophagus. 2008, 21: 9-14.PubMed Liu X, Li P, Zhang ST, You H, Jia JD, Yu ZL: COX-2 mRNA expression in esophageal squamous cell carcinoma (ESCC) and effect by NSAID. Dis Esophagus. 2008, 21: 9-14.PubMed
9.
go back to reference Buskens CJ, Van Rees BP, Sivula A, Reitsma JB, Haglund C, Bosma PJ, Offerhaus GJ, Van Lanschot JJ, Ristimäki A: Prognostic significance of elevated cyclooxygenase-2 expression in patients with adenocarcinoma of the esophagus. Gastroenterology. 2002, 122: 1800-1807. 10.1053/gast.2002.33580.CrossRefPubMed Buskens CJ, Van Rees BP, Sivula A, Reitsma JB, Haglund C, Bosma PJ, Offerhaus GJ, Van Lanschot JJ, Ristimäki A: Prognostic significance of elevated cyclooxygenase-2 expression in patients with adenocarcinoma of the esophagus. Gastroenterology. 2002, 122: 1800-1807. 10.1053/gast.2002.33580.CrossRefPubMed
10.
go back to reference Buttar NS, Wang KK, Leontovich O, Westcott J, Pacifico RJ, Anderson MA, Krishnadeth KK, Lutzke LS, Burgart LJ: Chemoprevention of esophageal adenocarcinoma by COX-2 inhibitors in an animal model of Barrett’s esophagus. Gastroenterology. 2002, 122: 1101-1112. 10.1053/gast.2002.32371.CrossRefPubMed Buttar NS, Wang KK, Leontovich O, Westcott J, Pacifico RJ, Anderson MA, Krishnadeth KK, Lutzke LS, Burgart LJ: Chemoprevention of esophageal adenocarcinoma by COX-2 inhibitors in an animal model of Barrett’s esophagus. Gastroenterology. 2002, 122: 1101-1112. 10.1053/gast.2002.32371.CrossRefPubMed
11.
go back to reference Greenhough A, Smartt HJ, Moore AE, Roberts HR, Williams AC, Paraskeva C, Kaidi A: The COX-2/PGE2 pathway: key roles in the hallmarks of cancer and adaptation to the tumour microenvironment. Carcinogenesis. 2009, 30: 377-386. 10.1093/carcin/bgp014.CrossRefPubMed Greenhough A, Smartt HJ, Moore AE, Roberts HR, Williams AC, Paraskeva C, Kaidi A: The COX-2/PGE2 pathway: key roles in the hallmarks of cancer and adaptation to the tumour microenvironment. Carcinogenesis. 2009, 30: 377-386. 10.1093/carcin/bgp014.CrossRefPubMed
12.
go back to reference Antonacopoulou AG, Tsamandas AC, Petsas T, Liava A, Scopa CD, Papavassiliou AG, Kalofonos HP: EGFR, HER-2 and COX-2 levels in colorectal cancer. Histopathology. 2008, 53: 698-706. 10.1111/j.1365-2559.2008.03165.x.CrossRefPubMed Antonacopoulou AG, Tsamandas AC, Petsas T, Liava A, Scopa CD, Papavassiliou AG, Kalofonos HP: EGFR, HER-2 and COX-2 levels in colorectal cancer. Histopathology. 2008, 53: 698-706. 10.1111/j.1365-2559.2008.03165.x.CrossRefPubMed
13.
go back to reference Brosens LA, Keller JJ, Pohjola L, Haglund C, Morsink FH, Iacobuzio-Donahue C, Goggins M, Giardiello FM, Ristimäki A, Offerhaus GJ: Increased expression of cytoplasmic HuR in familial adenomatous polyposis. Cancer Biol Ther. 2008, 7: 424-427. 10.4161/cbt.7.3.5417.CrossRefPubMed Brosens LA, Keller JJ, Pohjola L, Haglund C, Morsink FH, Iacobuzio-Donahue C, Goggins M, Giardiello FM, Ristimäki A, Offerhaus GJ: Increased expression of cytoplasmic HuR in familial adenomatous polyposis. Cancer Biol Ther. 2008, 7: 424-427. 10.4161/cbt.7.3.5417.CrossRefPubMed
14.
go back to reference Nosho K, Yoshida M, Yamamoto H, Taniguchi H, Adachi Y, Mikami M, Hinoda Y, Imai K: Association of Ets-related transcriptional factor E1AF expression with overexpression of matrix metalloproteinases, COX-2 and iNOS in the early stage of colorectal carcinogenesis. Carcinogenesis. 2005, 26: 892-899.CrossRefPubMed Nosho K, Yoshida M, Yamamoto H, Taniguchi H, Adachi Y, Mikami M, Hinoda Y, Imai K: Association of Ets-related transcriptional factor E1AF expression with overexpression of matrix metalloproteinases, COX-2 and iNOS in the early stage of colorectal carcinogenesis. Carcinogenesis. 2005, 26: 892-899.CrossRefPubMed
15.
go back to reference Loukanov T, Kirilov M, Fürstenberger G, Müller-Decker K: Localization of cyclo-oxygenase-2 in human recurrent colorectal cancer. Clin Invest Med. 2010, 33: E22-E29.PubMed Loukanov T, Kirilov M, Fürstenberger G, Müller-Decker K: Localization of cyclo-oxygenase-2 in human recurrent colorectal cancer. Clin Invest Med. 2010, 33: E22-E29.PubMed
16.
go back to reference Gustafsson A, Hansson E, Kressner U, Nordgren S, Andersson M, Wang W, Lönnroth C, Lundholm K: EP1-4 subtype, COX and PPAR gamma receptor expression in colorectal cancer in prediction of disease-specific mortality. Int J Cancer. 2007, 121: 232-240. 10.1002/ijc.22582.CrossRefPubMed Gustafsson A, Hansson E, Kressner U, Nordgren S, Andersson M, Wang W, Lönnroth C, Lundholm K: EP1-4 subtype, COX and PPAR gamma receptor expression in colorectal cancer in prediction of disease-specific mortality. Int J Cancer. 2007, 121: 232-240. 10.1002/ijc.22582.CrossRefPubMed
17.
go back to reference Delage B, Rullier A, Capdepont M, Rullier E, Cassand P: The effect of body weight on altered expression of nuclear receptors and cyclooxygenase-2 in human colorectal cancers. Nutr J. 2007, 6: 20-10.1186/1475-2891-6-20.CrossRefPubMedPubMedCentral Delage B, Rullier A, Capdepont M, Rullier E, Cassand P: The effect of body weight on altered expression of nuclear receptors and cyclooxygenase-2 in human colorectal cancers. Nutr J. 2007, 6: 20-10.1186/1475-2891-6-20.CrossRefPubMedPubMedCentral
18.
go back to reference Jahns F, Wilhelm A, Jablonowski N, Mothes H, Radeva M, Wölfert A, Greulich KO, Glei M: Butyrate suppresses mRNA increase of osteopontin and cyclooxygenase-2 in human colon tumor tissue. Carcinogenesis. 2011, 32: 913-920. 10.1093/carcin/bgr061.CrossRefPubMedPubMedCentral Jahns F, Wilhelm A, Jablonowski N, Mothes H, Radeva M, Wölfert A, Greulich KO, Glei M: Butyrate suppresses mRNA increase of osteopontin and cyclooxygenase-2 in human colon tumor tissue. Carcinogenesis. 2011, 32: 913-920. 10.1093/carcin/bgr061.CrossRefPubMedPubMedCentral
19.
go back to reference Pfaffl MW, Georgieva TM, Georgiev IP, Ontsouka E, Hageleit M, Blum JW: Real-time RT-PCR quantification of insulin-like growth factor (IGF)-1, IGF-1 receptor, IGF-2, IGF-2 receptor, insulin receptor, growth hormone receptor, IGF-binding proteins 1, 2 and 3 in the bovine species. Domest Anim Endocrinol. 2002, 22: 91-102. 10.1016/S0739-7240(01)00128-X.CrossRefPubMed Pfaffl MW, Georgieva TM, Georgiev IP, Ontsouka E, Hageleit M, Blum JW: Real-time RT-PCR quantification of insulin-like growth factor (IGF)-1, IGF-1 receptor, IGF-2, IGF-2 receptor, insulin receptor, growth hormone receptor, IGF-binding proteins 1, 2 and 3 in the bovine species. Domest Anim Endocrinol. 2002, 22: 91-102. 10.1016/S0739-7240(01)00128-X.CrossRefPubMed
20.
go back to reference Carvalho B, Sillars-Hardebol AH, Postma C, Mongera S, Droste JT, Obulkasim A, van de Wiel M, van Criekinge W, Ylstra B, Fijneman RJ, Meijer GA: Colorectal adenoma to carcinoma progression is accompanied by changes in gene expression associated with ageing, chromosomal instability, and fatty acid metabolism. Cell Oncol. 2012, 35: 53-63. 10.1007/s13402-011-0065-1.CrossRef Carvalho B, Sillars-Hardebol AH, Postma C, Mongera S, Droste JT, Obulkasim A, van de Wiel M, van Criekinge W, Ylstra B, Fijneman RJ, Meijer GA: Colorectal adenoma to carcinoma progression is accompanied by changes in gene expression associated with ageing, chromosomal instability, and fatty acid metabolism. Cell Oncol. 2012, 35: 53-63. 10.1007/s13402-011-0065-1.CrossRef
21.
go back to reference Hamaya Y, Yoshida K, Takai T, Ikuma M, Hishida A, Kanaoka S: Factors that contribute to faecal cyclooxygenase-2 mRNA expression in subjects with colorectal cancer. Br J Cancer. 2010, 102: 916-921. 10.1038/sj.bjc.6605564.CrossRefPubMedPubMedCentral Hamaya Y, Yoshida K, Takai T, Ikuma M, Hishida A, Kanaoka S: Factors that contribute to faecal cyclooxygenase-2 mRNA expression in subjects with colorectal cancer. Br J Cancer. 2010, 102: 916-921. 10.1038/sj.bjc.6605564.CrossRefPubMedPubMedCentral
22.
go back to reference Kanaoka S, Yoshida K, Miura N, Sugimura H, Kajimura M: Potential usefulness of detecting cyclooxygenase 2 messenger RNA in feces for colorectal cancer screening. Gastroenterology. 2004, 127: 422-427. 10.1053/j.gastro.2004.05.022.CrossRefPubMed Kanaoka S, Yoshida K, Miura N, Sugimura H, Kajimura M: Potential usefulness of detecting cyclooxygenase 2 messenger RNA in feces for colorectal cancer screening. Gastroenterology. 2004, 127: 422-427. 10.1053/j.gastro.2004.05.022.CrossRefPubMed
23.
go back to reference de Kok JB, Roelofs RW, Giesendorf BA, Pennings JL, Waas ET, Feuth T, Swinkels DW, Span PN: Normalization of gene expression measurements in tumor tissues: comparison of 13 endogenous control genes. Lab Invest. 2005, 85: 154-159. 10.1038/labinvest.3700208.CrossRefPubMed de Kok JB, Roelofs RW, Giesendorf BA, Pennings JL, Waas ET, Feuth T, Swinkels DW, Span PN: Normalization of gene expression measurements in tumor tissues: comparison of 13 endogenous control genes. Lab Invest. 2005, 85: 154-159. 10.1038/labinvest.3700208.CrossRefPubMed
24.
go back to reference Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT: The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009, 55: 611-622. 10.1373/clinchem.2008.112797.CrossRefPubMed Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, Mueller R, Nolan T, Pfaffl MW, Shipley GL, Vandesompele J, Wittwer CT: The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem. 2009, 55: 611-622. 10.1373/clinchem.2008.112797.CrossRefPubMed
25.
go back to reference Caradec J, Sirab N, Revaud D, Keumeugni C, Loric S: Is GAPDH a relevant housekeeping gene for normalisation in colorectal cancer experiments?. Br J Cancer. 2010, 103: 1475-1476. 10.1038/sj.bjc.6605851.CrossRefPubMedPubMedCentral Caradec J, Sirab N, Revaud D, Keumeugni C, Loric S: Is GAPDH a relevant housekeeping gene for normalisation in colorectal cancer experiments?. Br J Cancer. 2010, 103: 1475-1476. 10.1038/sj.bjc.6605851.CrossRefPubMedPubMedCentral
26.
go back to reference Barber RD, Harmer DW, Coleman RA, Clark BJ: GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 2005, 21: 389-395. 10.1152/physiolgenomics.00025.2005.CrossRefPubMed Barber RD, Harmer DW, Coleman RA, Clark BJ: GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues. Physiol Genomics. 2005, 21: 389-395. 10.1152/physiolgenomics.00025.2005.CrossRefPubMed
27.
go back to reference Stürzenbaum SR, Kille P: Control genes in quantitative molecular biological techniques: the variability of invariance. Comp Biochem Physiol B Biochem Mol Biol. 2001, 130: 281-289. 10.1016/S1096-4959(01)00440-7.CrossRefPubMed Stürzenbaum SR, Kille P: Control genes in quantitative molecular biological techniques: the variability of invariance. Comp Biochem Physiol B Biochem Mol Biol. 2001, 130: 281-289. 10.1016/S1096-4959(01)00440-7.CrossRefPubMed
28.
go back to reference Bustin SA: Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol. 2000, 25: 169-193. 10.1677/jme.0.0250169.CrossRefPubMed Bustin SA: Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol. 2000, 25: 169-193. 10.1677/jme.0.0250169.CrossRefPubMed
Metadata
Title
Over-expression of COX-2 mRNA in colorectal cancer
Authors
Hennie MJ Roelofs
Rene HM te Morsche
Bjorn WH van Heumen
Fokko M Nagengast
Wilbert HM Peters
Publication date
01-12-2014
Publisher
BioMed Central
Published in
BMC Gastroenterology / Issue 1/2014
Electronic ISSN: 1471-230X
DOI
https://doi.org/10.1186/1471-230X-14-1

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