J Korean Surg Soc.  2011 Aug;81(2):115-121. 10.4174/jkss.2011.81.2.115.

Up-regulation of cyclooxygenase-2-derived prostaglandin E2 in colon cancer cells resistant to 5-fluorouracil

• Choi CH
• Lee TB
• Lee YA
• Choi S
• Kim KJ

• KMID: 2096665
• DOI: http://doi.org/10.4174/jkss.2011.81.2.115

Abstract

PURPOSE
It has been suggested that constitutive up-regulation of cyclooxygenase (COX)-2 is associated with resistance to apoptosis, increased angiogenesis, and increased tumor invasiveness in various cancers including colon cancer. There are many factors involved in the resistance to 5-fluorouracil (5-FU) in colon cancer. However, little is known about the role of COX-2 in acquired resistance to 5-FU in colon cancer.
METHODS
Hence we investigated whether COX-2 contribute to acquired resistance to 5-FU in colon cancer cells, using cytotoxicity assay for cell survival, reverse transcription-polymerase chain reaction (RT-PCR) for vascular endothelial growth factor (VEGF), quantitative RT-PCR for COX-1 and COX-2, and enzyme-linked immunosorbent assay for PGE2.
RESULTS
The 5-FU resistant colon cancer cells, SNU-C5/5FUR, showed increased expression of COX-2, prostaglandin E2 (PGE2), and VEGF, compared to its parental cell (SNU-C5). By treatment with meloxicam, the expression of PGE2 and VEGF was reduced significantly in the resistant cells, but not in the parent cells.
CONCLUSION
These results demonstrate that COX-2 derived PGE2 is up-regulated and COX-2 inhibitor may have an anti-angiogenic effect in the colon cancer cells resistant to 5-FU.

Keyword

Cyclooxygenase-2; Prostaglandin E2; 5-Fluorouracil; Acquired resistance; Colon neoplasms

MeSH Terms

Apoptosis
Cell Survival
Colon
Colonic Neoplasms
Cyclooxygenase 2
Dinoprostone
Enzyme-Linked Immunosorbent Assay
Fluorouracil
Humans
Parents
Prostaglandin-Endoperoxide Synthases
Thiazines
Thiazoles
Up-Regulation
Vascular Endothelial Growth Factor A
Cyclooxygenase 2
Dinoprostone
Fluorouracil
Prostaglandin-Endoperoxide Synthases
Thiazines
Thiazoles
Vascular Endothelial Growth Factor A

Figure

• 

  Fig. 1 mRNA expression patterns of cyclooxygenase (COX)-2 in SNU-C5 and SNU-C5/5FUR determined by quantitative reverse transcription- polymerase chain reaction. (A) Significant difference in the ratio of COX-2/β-actin between SNU-C5 and SNU-C5/5FUR (P = 0.000727)a), (B) Significant difference in the ratio COX-2/COX-1 between SNU-C5 and SNU-C5/5FUR (P = 0.00117).a)

• 

  Fig. 2 Comparison of prostaglandin E2 (PGE2) levels between SNU-C5 and SNU-C5/5FUR. PGE2 levels present in the supernatant of the culture medium were determined by enzyme-linked immunosorbent assay using PGE2 monoclonal antibody. a)Compare PGE2 levels between SNU-C5 and SNU-C5/5FUR at basal state. b)Compare PGE2 levels after 12 hours treatment of meloxicam (10 µM) with control in SNU-C5/5FUR. c)Compare PGE2 levels after meloxicam (100 µM) with control in SNU-C5/5FUR, all of which have P-values less than 0.01.

• 

  Fig. 3 Cytotoxic effect of 5-fluorouracil (5-FU) alone and/or in combination with meloxicam in SNU-C5 (A) and SNU-C5/5FUR (B). 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay was used to analyze cell viability.

• 

  Fig. 4 Expression patterns of vascular endothelial growth factor (VEGF) mRNA in SNU-C5 and SNU-C5/5FUR at basal state and after treatment of meloxicam (10, 100 and 500 µM) for 24 hours by transcription-polymerase chain reaction method.

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