Cancer Research Meeting Calendar Jordan
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gupta, R. A.
Right arrow Articles by DuBois, R. N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gupta, R. A.
Right arrow Articles by DuBois, R. N.
[Cancer Research 63, 906-911, March 1, 2003]
© 2003 American Association for Cancer Research

Advances in Brief

Cyclooxygenase-1 is Overexpressed and Promotes Angiogenic Growth Factor Production in Ovarian Cancer1

Rajnish A. Gupta2, Lovella V. Tejada2, Beverly J. Tong3, Sanjoy K. Das, Jason D. Morrow, Sudhansu K. Dey4 and Raymond N. DuBois4

Departments of Medicine [R. A. G., J. D. M., R. N. D.], Cell and Developmental Biology [R. A. G., S. K. Dey., R. N. D.], Pediatrics [L. V. T., S. K. Das., S. K. Dey.], and Pharmacology [J. D. M.], Vanderbilt University School of Medicine, Nashville, Tennessee 37232


   ABSTRACT
Top
 ABSTRACT
Introduction
 Materials and Methods
 Results and Discussion
 REFERENCES
 
Inhibition of cyclooygenase-2 (COX-2) catalytic activity has proven successful in restricting the growth of epithelial-derived cancers in vivo. Whether COX-2 inhibitor therapy would be beneficial in the prevention and/or treatment of ovarian cancer, the most lethal gynecological malignancy worldwide, is not known. Most patients with ovarian cancer undergo cytoreductive therapy. Because many of the cytotoxic drugs used to treat ovarian cancer induce COX-2 expression, samples from patients that had not undergone cytoreductive therapy were specifically chosen for COX isoform expression analysis. A majority of specimens exhibited elevated levels of COX-1, not COX-2, mRNA, and protein compared with normal ovarian tissue. Focal regions within the tumor expressing high COX-1 also had elevated levels of pro-angiogenic proteins. Selective inhibition of COX-1, not COX-2, inhibited arachidonic acid-stimulated vascular endothelial growth factor production, which could be reversed by cotreatment with prostaglandin E2. Thus, COX-1 may contribute to carcinoma development in the ovary through stimulation of neovascularization. Clinical studies testing the efficacy of COX inhibition as adjuvant therapy for ovarian cancer may see more beneficial effects with adjuvant therapy with either a COX-1 selective or nonselective cyclooxygenase inhibitor as compared with a COX-2 selective drug.


   Introduction
Top
 ABSTRACT
 Introduction
Materials and Methods
 Results and Discussion
 REFERENCES
 
The COX5 enzymes COX-1 and COX-2 catalyze the rate-limiting step in the biosynthesis of PGs derived from AA. Research over the last decade, primarily in studies focused on colorectal cancer, has established that NSAIDs are effective in both cancer prevention and as adjuvant therapy in the treatment of established tumors (1) . These drugs are thought to inhibit colorectal cancer cell growth primarily through inhibition of COX-2, although other noncyclooxygenase biochemical targets may be involved. Recent experiments suggest that COX-2 is up-regulated in a range of extracolonic cancers, and selective COX-2 inhibitors have potent antineoplastic effects in vivo in preclinical models of a variety of solid malignancies. These data have led to the initiation of a number of clinical trials that are testing the efficacy of COX-2-specific inhibitor therapy in the primary or secondary prevention of cancer or as part of a combination therapy regimen for established tumors. Whether COX-2 selective inhibitor therapy will prove beneficial in the prevention and/or treatment of ovarian cancer is not known. Ovarian cancer is associated with a high mortality rate because of the absence of effective screening strategies to identify patients at high risk or who have already developed early neoplastic lesions still amenable to treatment (2 , 3) .

In addition, treatment options for patients diagnosed with advanced disease remain inadequate. Thus, ovarian cancer represents a potential candidate to target for chemoprevention. The results of epidemiological studies examining whether NSAIDs can prevent or delay the development of ovarian cancer have been mixed. Several population- and hospital-based case control studies have documented that exposure to several NSAIDs is associated with a time- and dose-dependent decrease in the risk for the development of ovarian cancer (4, 5, 6) , whereas other studies detected either no statistically significant association or found the reduction in risk to be associated only with the use of particular NSAIDs (7 , 8) . There are also conflicting data available describing the expression of COX isotypes in ovarian cancer; Dore et al. (9) found COX-1 to be the predominant COX isoform expressed, whereas two other groups reported finding high levels of COX-2 (10 , 11) . However, these latter two expression studies did not specifically rule out samples that had undergone cytoreductive therapy, a treatment that is common in patients presenting with advanced ovarian malignancy. Because many compounds used in this type of therapy induce COX-2 (12 , 13) , it is unclear if COX-2 was found to be elevated because of the malignant process itself or simply secondary to the use of cytotoxic agents for treatment of primary disease. To investigate the role of the cyclooxygenase pathway in the pathogenesis of ovarian cancer and determine whether a scientific rationale exists for the use of COX inhibitors in the prevention and/or treatment of the disease, we evaluated COX-1 and COX-2 expression in ovarian cancer samples taken specifically from patients who had not undergone cytoreductive therapy.


   Materials and Methods
Top
 ABSTRACT
 Introduction
 Materials and Methods
Results and Discussion
 REFERENCES
 
Cell Culture.
The SK-OV-3, OV90, and OVCAR-3 cells were purchased from American Type Culture Collection and grown in the recommended media under standard conditions.

Tissue Samples.
Tissue samples of 11 epithelial ovarian cancers that had not undergone cytoreductive therapy and nine normal human ovaries were obtained immediately after surgery from surgical pathology specimens (University of Kansas Medical Center and Vanderbilt University). The tissues obtained from the pathologist were flash frozen in liquid Histo-freeze (Fisher) and stored at -80°C.

RNA Isolation and Northern Blot Analysis.
Total RNA was extracted from the tissue specimens using TRIzol Kit (Life Technologies, Inc.). Total RNA (20 µg) was denatured, separated by formaldehyde-agarose gel electrophoresis, transferred, and cross-linked to nylon membranes by UV irradiation. Northern blots were prehybridized, hybridized, and washed as described previously (14 , 15) . For Northern hybridization, antisense 32P-labled cRNA probes for mouse COX-1, human COX-2, and ß-actin were generated. After hybridization, the blots were washed under stringent conditions, and the hybrids were detected by autoradiography. Stripping of the hybridized probe before subsequent rehybridization was achieved. Each blot was hybridized sequentially to COX-2, COX-1, and ß-actin probes.

In Situ Hybridization.
In situ hybridization followed the protocol described previously (14) . Frozen sections (10 µM) from each tissue specimen were mounted onto poly-L-lysine-coated slides and stored at -80°C until used. Serial sections were obtained to detect localization of gene expression in similar areas. Sections were brought to room temperature, fixed in cold 4% paraformaldehyde solution in PBS, acetylated, and hybridized at 45°C for 4 h in 50% formamide buffer containing 35S-labeled antisense or sense cRNA probes specific to mouse cRNA to COX-1 and human-specific cRNAs to COX-2, HIF-1{alpha}, VEGF, and Flk-1. After hybridization and washing, the slides were incubated with RNase A (20 µg/ml) at 37°C for 20 min, and RNase A-resistant hybrids were detected by autoradiography using Kodak NTB-2 liquid emulsion. Parallel sections hybridized with the sense probes served as negative controls. Slides were poststained with H&E.

Immunohistochemistry.
Immunocolocalization of COX-1 and COX-2 was performed in 2% paraformaldehyde-fixed frozen sections using a Zymed-Histostain SP kit (Zymed). Rabbit antipeptide antibodies to mouse COX-1 and COX-2 were used as described previously (15) . These antibodies were found to cross-react with human tissues. After immunostaining, sections were lightly counterstained with hematoxylin or fast green. Red deposits indicate the site of immunoreactive proteins.

Preparation of Tissue Lysates.
Tissue samples were homogenized in lysis buffer [1% Triton X-100, 1% deoxycholate, 10 mM Tris (pH 7.2), 150 mM NaCl, 1 mM sodium vanadate, 50 mM sodium fluoride, 1 mM phenylmetheysulfonyl fluoride, 50 mg/ml aprotinin, and 50 mg/ml leupeptin]. The samples were transferred to eppendorf tubes and centrifuged at 14,000 x g for 15 min in the cold. The supernatants were transferred to fresh tubes, and protein concentrations were determined. These extracts were used for Western blot analysis.

Western Blot Analysis.
Lysates (50 µg of protein/lane) were analyzed by SDS-PAGE on 10% Tris-glycine gels. Protein was electrotransferred to nitrocellulose membranes and blocked with a solution of PBS containing 5% milk and 0.1% Tween 20. Bands were detected using chemiluminescent detection reagents (Pierce). Blots were probed with a goat polyclonal antibody against COX-1 or COX-2 (Santa Cruz Biotechnology) followed by a peroxidase-conjugated antigoat (Sigma) or donkey-antigoat (Jackson ImmunoResearch Laboratories), respectively. After incubation, antibodies were washed in PBS and 0.1% Tween 20. Bands were detected using chemiluminescent detection reagents (Pierce).

PG Measurements.
PG profiles for the OVCAR-3 cells were measured and quantified using a gas chromatography/negative ion chemical ionization mass spectrometric assay and a PGE2 immunoassay kit (R&D Systems).

Cell Growth Measurements.
Cells were plated at a density of ~20%, and treatment was initiated the following day with 0.1% DMSO or the indicated dose of COX inhibitor. Fresh media and drug was added every 48 h, and the number of viable cells was determined using a Coulter counter after 7 days of treatment.

VEGF Measurements.
VEGF concentrations in the media of OVCAR-3 cells treated with the indicated dose of AA, COX inhibitor, and PGE2 were measured using the Quantikine Human VEGF Immunoassay kit (R&D Systems) according to manufacturer instructions.


   Results and Discussion
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results and Discussion
REFERENCES
 
COX-1 and COX-2 expression levels in 11 epithelial ovarian cancers and nine normal human ovaries were determined using Northern blot and immunoblot analysis. Significant levels of COX-1 or COX-2 mRNA and protein were not detected in any of the normal human ovarian tissue (Fig. 1, A and B)Citation . However, dramatic elevations of COX-1, not COX-2, protein and mRNA were detected in a majority of the ovarian cancer samples tested. In colorectal and other cancers, COX-1 and COX-2 protein levels have been determined in multiple cell types, including epithelial, fibroblast, macrophage, and endothelial cells. In situ hybridization and immunohistochemistry analysis were done to determine the cellular localization of COX-1 and COX-2 protein in ovarian cancer. A significant elevation in COX-1, not COX-2, mRNA was observed by in situ hybridization, and COX-1 mRNA was localized to the epithelial compartment of the tumors (Fig. 2, A and B)Citation . Immunohistochemical analysis of sections of both normal human ovary and ovarian epithelial cancers confirmed the elevated expression of COX-1, not COX-2, protein in ovarian tumors, and the immunoreactivity detected with a COX-1-specific antibody was localized primarily to ovarian carcinoma cells within the tumor (Fig. 2C)Citation .



View larger version (55K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 1. COX-1, not COX-2, mRNA and protein levels are elevated in human ovarian cancer. A, Northern hybridization. Total RNA (20 µg) from normal human ovary and ovarian epithelial tumor tissue was fractionated on a formaldehyde-agarose gel and probed for COX-1 and COX-2 and ß-actin mRNA expression. B, Western blotting. Total protein (50 µg) from normal human ovary and ovarian tumor tissue was fractionated on a 10% SDS-PAGE and probed with goat polyclonal antibodies specific for COX-1 and COX-2. The positive control lane labeled C consists of recombinant COX-1 and COX-2 protein, whereas the positive control lane C* consists of an aliquot of cell lysate from the HCA-7 rectal adenocarcinoma line known to express high levels of COX-2 protein.

 


View larger version (91K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 2. COX-1, not COX-2, mRNA is expressed and localized to the epithelial compartment in human ovarian cancer. In A and B, COX-1 and COX-2 mRNA localization was determined using in situ hybridization in two different pairs of normal human ovary and ovarian cancer. Parallel sections hybridized with sense cRNA COX-1 and COX-2 probes served as negative controls (data not shown). In C, COX-1 and COX-2 protein expression and cellular localization were determined by immunohistochemistry using rabbit antipeptide antibodies to COX-1 and COX-2.

 
Angiogenesis, defined as the generation of new capillaries from preexisting vessels, is a critical factor in the sustained growth of solid tumors. Ovarian cancer is known to be highly vascular and is a primary cancer in which current antiangiogenic therapies are being tested (16 , 17) . VEGF and the VEGF receptor flk-1 are highly expressed in a majority of ovarian epithelial tumors, and VEGF expression is a negative prognostic factor for the disease (18 , 19) . The transcription factor HIF-1{alpha} is a dominant regulator of VEGF gene transcription and induces significant increases in VEGF mRNA copy number in response to various stimuli by binding to a hypoxia-responsive element within the VEGF promoter (20) . There is also a strong link between the COX pathway and angiogenesis. Data from multiple groups suggest that a major mechanism by which COX-derived PGs promote polyp growth in the colon is through the stimulation of new blood vessel growth. To determine whether regions within ovarian tumors demonstrating high COX-1 expression correlate with foci of prominent angiogenic activity, in situ hybridization was done probing for COX-1, VEGF, Flk-1, and HIF-1{alpha} in serial sections. Regions of ovarian epithelial cells exhibiting high COX-1 also expressed significant levels of HIF-1{alpha} and VEGF (Figs. 2ACitation and 3, A and BCitation ). High levels of flk-1 were seen in the endothelial cells located in the stroma adjacent to ovarian epithelial cells expressing COX-1, HIF-1{alpha}, and VEGF (Fig. 3C)Citation . The above data suggest that elevations in COX-1 expression are enhanced in regions of ovarian epithelial tumors undergoing extensive angiogenesis.



View larger version (78K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 3. The expression of VEGF, HIF-1{alpha}, and Flk1 mRNA correlates with COX-1 expression in human ovarian cancer. In situ hybridization of serial sections was performed to determine the expression of the proangiogenic factors VEGF (A), HIF-1{alpha} (B), and Flk1 (C) in regions of human ovarian tumors expressing high levels of COX-1 mRNA (see Fig. 2ACitation ). Parallel sections hybridized with sense cRNA probes for all three genes served as negative controls (data not shown).

 
To directly test the hypothesis that COX-1-derived PGs promote the development of ovarian cancer, a cell culture-based model system was developed and evaluated. The established ovarian cancer cell lines SK-OV-3, OV90, and OVCAR-3 were evaluated for COX-1 and COX-2 protein levels. Although none of the three cell lines expressed detectable levels of COX-2, the OVCAR-3 cells exhibited high levels of COX-1 protein and were chosen for further study (Fig. 4A)Citation . No reports have been published documenting the types of PGs synthesized by ovarian carcinoma cells. Levels of the five major PG metabolites were measured in these cells in response to stimulation with AA, and significant levels of PGF2{alpha}, PGI2, PGE2, and PGD2 were detected (Fig. 4B)Citation . Although most NSAIDs inhibit the enzymatic activity of both COX-1 and COX-2, isoform selective COX inhibitors have been developed. SC-560 is 700 times more selective for inhibiting COX-1 compared with COX-2, whereas celecoxib is ~3000-fold more selective for COX-2 versus COX-1 (21) . The ability of increasing doses of either SC-560 or celecoxb to inhibit AA-induced PGE2 formation in OVCAR-3 cells was evaluated. SC-560, but not celecoxib, inhibited PGE2 formation in a dose-dependent manner (Fig. 4C)Citation . These data confirmed our expression studies, indicating that COX-1 is the major source of PG production in these cells.



View larger version (29K):
[in this window]
[in a new window]
[Download PPT slide]
  		Fig. 4. The OVCAR-3 human ovarian epithelial cancer cell line expresses high levels of catalytically active COX-1, and COX-1-derived PGE2 stimulates VEGF secretion in the OVCAR-3 human ovarian cancer cell line. In A, whole-cell lysates from the SK-OV-3, OV90, and OVCAR-3 human ovarian epithelial cancer cell lines were fractionated on a 10% PAGE and probed with goat polyclonal antibodies specific for COX-1 and COX-2. The control lane for the COX-1 blot is an aliquot of protein lysate from the MC-26 breast cancer cell line known to express high levels of COX-1, whereas the control lane for the COX-2 blot is an aliquot of protein lysate from the HCA-7 rectal adenocarcinoma line known to express high levels of COX-2 protein. In B, OVCAR-3 cells grown in serum-depleted media were treated with AA (20 µM) for 4 h, followed by measurement of different PG subtypes using a gas chromatography/negative ion chemical ionization mass spectrometric assay. In C, OVCAR-3 cells grown in serum-depleted media were treated with AA (20 µM) and increasing concentrations of a selective inhibitor against COX-1 (SC-560) or a selective inhibitor of COX-2 (celecoxib), followed by measurement of PGE2 levels using a PGE2 immunoassay. In D, OVCAR-3 cells were treated with increasing doses of a COX-1 selective (SC-560), non-COX isotype selective (indomethacin), or COX-2 selective (celecoxib) inhibitor for 7 days after the number of viable cells was counted and expressed as a percentage of control-treated cells (0.1% DMSO). Each experiment was done in triplicate, and each data point represents the mean of two independent experiments. Error bars = SE. In E, OVCAR-3 cells grown in serum-depleted media were treated with AA (20 µM) and the indicated concentrations of a COX-1 (SC-560) or COX-2 (celecoxib) selective inhibitor with or without the indicated dose of PGE2 for 24 h, after which, VEGF levels in the media were measured using a human VEGF immunoassay.

 
Finally, the ability of SC-560 to inhibit either DNA replication or VEGF secretion was tested to determine the functional relevance of COX-1 expression in ovarian cancer epithelial cells. Treatment of OVCAR-3 cells with increasing doses of either SC-560, celecoxib, or the non-COX isotype selective NSAID indomethacin resulted in significant decreases in cell number only at doses > 50 µM (Fig. 4D)Citation . Biological effects seen at such high drug concentrations are unlikely to be caused by inhibition of COX catalytic activity and have limited clinical relevance, because these levels are 50–100 times greater than the concentration of COX inhibitors that can be achieved in vivo (22) . In contrast, treatment of OVCAR-3 cells with a low dose of SC-560, but not celecoxib, significantly inhibited AA-induced VEGF secretion, and this effect could be partially reversed by cotreatment with PGE2 (Fig. 4E)Citation .

Neoplasms of the ovary rarely produce symptoms that lead to prompt medical attention until the disease is highly advanced and often incurable. Currently, no effective detection strategies exist to identify patients at high risk or who have localized lesions that can be effectively treated. These facts make ovarian cancer account for a disproportionate number of lethal cancers and thus a primary candidate to target for prevention. Given the effectiveness of NSAID therapy in the chemoprevention of colorectal cancer, as well as several other types of solid tumors, it is important to determine the role, if any, that NSAID treatment may have in the management of ovarian cancer. Our results here suggest that: (a) the COX-1 enzyme is overexpressed in a significant number of ovarian cancers; (b) COX-1 may promote ovarian cancer development via stimulation of angiogenesis; and (c) COX-1 or non-COX selective inhibitors should be further evaluated for their ability to inhibit ovarian cancer cell growth.

Our results indicate that COX-1, not COX-2, mRNA and protein levels are elevated in ovarian cancers from patients not exposed previously to cytotoxic chemotherapy. Similar results were obtained by Dore et al. (9) , who used immunohistochemistry to demonstrate strong expression of COX-1, not COX-2, protein in human ovarian cancer specimens. Both of these results are in contradiction to data generated by two other groups that report elevated COX-2 levels in ovarian cancer (10 , 11) . However, neither of these two groups determined the status of COX-1 expression in their samples. The reason for the discrepancy with regard to COX-2 expression is not known but may be related to differences in clinical treatment regimens of patients before tissue collection, detection methods, tissue processing, and/or antibody cross-reactivity.

A majority of epithelial-derived tumors with elevated PG levels overexpresses COX-2 and not COX-1. This is consistent with the hypothesis that COX-1 is constitutively expressed and responsible for basal PG production, whereas COX-2 is highly inducible and responsible for the elevations in PG production that occur in response to pro-inflammatory cytokines and growth factors, both of which are likely to be highly concentrated within the microenvironment of a tumor. The basis for the divergent expression patterns of COX-1 and COX-2 in ovarian cancer is not known. The simplest explanation may be that the development of ovarian cancer is associated with the dysregulation of a unique combination of signaling pathways not found in other tumor types that converge to cause activation of COX-1, not COX-2, gene transcription. The COX-1 promoter is relatively uncharacterized, and transfection experiments using COX-1 and COX-2 promoter–reporter constructs in the OVCAR-3 cells may help determine the molecular basis for why COX-1 and not COX-2 is up-regulated ovarian cancer.

The role of COX-1 in neoplasia is not clear. COX-1 and COX-2 catalyze identical biochemical reactions. It is thus possible that the elevated COX-2 seen in many types of cancer is selected to simply increase total PG levels within the tumor microenvironment. Alternatively, COX-2 may exhibit unique substrate utilization, cellular or subcellular localization, and/or coupling with downstream PG synthases, resulting in a pro-oncogenic effect that cannot be substituted by COX-1. There are published reports using genetically modified mice to support both theories. Chulada et al. (23) demonstrated an equivalent reduction in intestinal polyposis in Min/+ mice with a genetic disruption of COX-1 or COX-2, and both genes were found to contribute equally to PGE2 levels within polyps. In contrast, Williams et al. (24) found that xenografts of Lewis lung carcinoma cells grew more slowly only in mice genetically null for COX-2 but not COX-1. Future experiments using the OVCAR-3 cells may help clarify whether COX-1 has a unique pro-tumorigenic role in ovarian cancer or if it is simply an issue of generating a threshold level of PGs that can be derived from either COX isotype, e.g., mRNA antisense or RNAi could be used to inhibit COX-1 expression (and hence VEGF secretion) in these cells and rescue experiments performed to determine whether transfection with COX-2 can substitute for COX-1 to stimulate angiogenic growth factor production.

Our results imply that PGE2 positively regulates VEGF production in ovarian epithelial cells perhaps via the H1F1-{alpha} pathway. Future experiments designed to determine the expression levels of all four PGE2 receptor (EP) subtypes in human ovarian cancer, as well as the ability of EP receptor subtype-specific agonists to stimulate angiogenic growth factor production, will be important.

In summary, our current results suggest that use of COX-1 or non-COX isotype selective NSAIDs may be more plausible than COX-2 inhibitor therapy in either the primary or secondary prevention of ovarian cancer or as a component of a cancer treatment for advanced tumors. However, it will be important to test this hypothesis in vivo using COX inhibitors that preferentially inhibit COX-1 or COX-2, and such experiments using preclinical models of ovarian cancer cell growth are currently underway.


   FOOTNOTES
 
The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1 Supported in part from USPHS Grants HD37830 (S. K. D.), HD12304, and HD33994 and Mellon Foundation (S. K. D.) Grants DK 47297, DK 62112, and PO1CA-77839 (R. N. D.). S. K. D. is a recipient of an National Institute of Child Health and Human Development/NIH MERIT Award. R. N. D. is the Mina C. Wallace Professor of Cancer Prevention. Back

2 R. A. G. and L. V. T. contributed equally to this work. Back

3 Present address: Heartland Women’s Health Care, 200 North East 54th Street, Kansas City, MO 64118. Back

4 To whom correspondence should be addressed, at sk.dey{at}vanderbilt.edu (S. K. D.) or raymond.dubois{at}vanderbilt.edu (R. N. D.). Back

5 The abbreviations used are: COX, cyclooxygenase; NSAID, nonsteroidal anti-inflammatory drug; PG, prostaglandin; AA, arachidonic acid; HIF, hypoxia-inducible factor; VEGF, vascular endothelial growth factor. Back

Received 11/20/02. Accepted 1/20/03.


   REFERENCES
Top
 ABSTRACT
 Introduction
 Materials and Methods
 Results and Discussion
 REFERENCES
 

  1. Gupta R. A., DuBois R. N. Colorectal cancer prevention and treatment by inhibition of cyclooxygenase-2. Nat. Rev. Cancer, 1: 11-21, 2001.[Medline]
  2. Ozols R. F. Future directions in the treatment of ovarian cancer. Semin. Oncol., 29: 32-42, 2002.
  3. NIH consensus conference. Ovarian cancer. Screening, treatment, and follow-up. NIH Consensus Development Panel on Ovarian Cancer. JAMA, 273: 491-497, 1995.[Abstract/Free Full Text]
  4. Cramer D. W., Harlow B. L., Titus-Ernstoff L., Bohlke K., Welch W. R., Greenberg E. R. Over-the-counter analgesics and risk of ovarian cancer. Lancet, 351: 104-107, 1998.[Medline]
  5. Moysich K. B., Mettlin C., Piver M. S., Natarajan N., Menezes R. J., Swede H. Regular use of analgesic drugs and ovarian cancer risk. Cancer Epidemiol. Biomark. Prev., 10: 903-906, 2001.[Abstract/Free Full Text]
  6. Akhmedkhanov A., Toniolo P., Zeleniuch-Jacquotte A., Kato I., Koenig K. L., Shore R. E. Aspirin and epithelial ovarian cancer. Prev. Med., 33: 682-687, 2001.[Medline]
  7. Rosenberg L., Palmer J. R., Rao R. S., Coogan P. F., Strom B. L., Zauber A. G., Stolley P. D., Shapiro S. A case-control study of analgesic use and ovarian cancer. Cancer Epidemiol. Biomark. Prev., 9: 933-937, 2000.[Abstract/Free Full Text]
  8. Tavani A., Gallus S., La Vecchia C., Conti E., Montella M., Franceschi S. Aspirin and ovarian cancer: an Italian case-control study. Ann. Oncol., 11: 1171-1173, 2000.[Abstract/Free Full Text]
  9. Dore M., Cote L. C., Mitchell A., Sirois J. Expression of prostaglandin G/H synthase type 1, but not type 2, in human ovarian adenocarcinomas. J. Histochem. Cytochem., 46: 77-84, 1998.[Abstract/Free Full Text]
  10. Klimp A. H., Hollema H., Kempinga C., van der Zee A. G., de Vries E. G., Daemen T. Expression of cyclooxygenase-2 and inducible nitric oxide synthase in human ovarian tumors and tumor-associated macrophages. Cancer Res., 61: 7305-7309, 2001.[Abstract/Free Full Text]
  11. Matsumoto Y., Ishiko O., Deguchi M., Nakagawa E., Ogita S. Cyclooxygenase-2 expression in normal ovaries and epithelial ovarian neoplasms. Int. J. Mol. Med., 8: 31-36, 2001.[Medline]
  12. Cassidy P. B., Moos P. J., Kelly R. C., Fitzpatrick F. A. Cyclooxygenase-2 induction by paclitaxel, docetaxel, and taxane analogues in human monocytes and murine macrophages: structure-activity relationships and their implications. Clin. Cancer Res., 8: 846-855, 2002.[Abstract/Free Full Text]
  13. Subbaramaiah K., Hart J. C., Norton L., Dannenberg A. J. Microtubule-interfering agents stimulate the transcription of cyclooxygenase-2. Evidence for involvement of ERK1/2 AND p38 mitogen-activated protein kinase pathways. J. Biol. Chem., 275: 14838-14845, 2000.[Abstract/Free Full Text]
  14. Das S. K., Wang X. N., Paria B. C., Damm D., Abraham J. A., Klagsbrun M., Andrews G. K., Dey S. K. Heparin-binding EGF-like growth factor gene is induced in the mouse uterus temporally by the blastocyst solely at the site of its apposition: a possible ligand for interaction with blastocyst EGF-receptor in implantation. Development, 120: 1071-1083, 1994.[Abstract]
  15. Chakraborty I., Das S. K., Wang J., Dey S. K. Developmental expression of the cyclo-oxygenase-1 and cyclo-oxygenase-2 genes in the peri-implantation mouse uterus and their differential regulation by the blastocyst and ovarian steroids. J. Mol. Endocrinol., 16: 107-122, 1996.[Abstract/Free Full Text]
  16. Hazelton D. A., Hamilton T. C. Vascular endothelial growth factor in ovarian cancer. Curr. Oncol. Rep., 1: 59-63, 1999.[Medline]
  17. Abulafia O., Triest W. E., Sherer D. M. Angiogenesis in malignancies of the female genital tract. Gynecol. Oncol., 72: 220-231, 1999.[Medline]
  18. Yamamoto S., Konishi I., Mandai M., Kuroda H., Komatsu T., Nanbu K., Sakahara H., Mori T. Expression of vascular endothelial growth factor (VEGF) in epithelial ovarian neoplasms: correlation with clinicopathology and patient survival, and analysis of serum VEGF levels. Br. J. Cancer, 76: 1221-1227, 1997.[Medline]
  19. Paley P. J., Staskus K. A., Gebhard K., Mohanraj D., Twiggs L. B., Carson L. F., Ramakrishnan S. Vascular endothelial growth factor expression in early stage ovarian carcinoma. Cancer, 80: 98-106, 1997.[Medline]
  20. Forsythe J. A., Jiang B. H., Iyer N. V., Agani F., Leung S. W., Koos R. D., Semenza G. L. Activation of vascular endothelial growth factor gene transcription by hypoxia-inducible factor 1. Mol. Cell. Biol., 16: 4604-4613, 1996.[Abstract]
  21. Smith C. J., Zhang Y., Koboldt C. M., Muhammad J., Zweifel B. S., Shaffer A., Talley J. J., Masferrer J. L., Seibert K., Isakson P. C. Pharmacological analysis of cyclooxygenase-1 in inflammation. Proc. Natl. Acad. Sci. USA, 95: 13313-13318, 1998.[Abstract/Free Full Text]
  22. Williams C. S., Watson A. J., Sheng H., Helou R., Shao J., DuBois R. N. Celecoxib prevents tumor growth in vivo without toxicity to normal gut: lack of correlation between in vitro and in vivo models. Cancer Res., 60: 6045-6051, 2000.[Abstract/Free Full Text]
  23. Chulada P. C., Thompson M. B., Mahler J. F., Doyle C. M., Gaul B. W., Lee C., Tiano H. F., Morham S. G., Smithies O., Langenbach R. Genetic disruption of Ptgs-1, as well as Ptgs-2, reduces intestinal tumorigenesis in Min mice. Cancer Res., 60: 4705-4708, 2000.[Abstract/Free Full Text]
  24. Williams C. S., Tsujii M., Reese J., Dey S. K., DuBois R. N. Host cyclooxygenase-2 modulates carcinoma growth. J. Clin. Investig., 105: 1589-1594, 2000.[Medline]



This article has been cited by other articles:


Home page
 ReproductionHome page
H. N Jabbour, K. J Sales, R. D Catalano, and J. E Norman
Inflammatory pathways in female reproductive health and disease
Reproduction, December 1, 2009; 138(6): 903 - 919.
[Abstract] [Full Text] [PDF]

Home page
 Neuro Oncol DukeHome page
S. Phuphanich, K. A. Carson, S. A. Grossman, G. Lesser, J. Olson, T. Mikkelsen, S. Desideri, J. D. Fisher, and for the New Approaches to Brain Tumor Therapy (NAB
Phase I safety study of escalating doses of atrasentan in adults with recurrent malignant glioma
Neuro-oncol, August 1, 2008; 10(4): 617 - 623.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
R. S. Freedman, E. Wang, S. Voiculescu, R. Patenia, R. L. Bassett Jr., M. Deavers, F. M. Marincola, P. Yang, and R. A. Newman
Comparative Analysis of Peritoneum and Tumor Eicosanoids and Pathways in Advanced Ovarian Cancer
Clin. Cancer Res., October 1, 2007; 13(19): 5736 - 5744.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
T. Daikoku, S. Tranguch, A. Chakrabarty, D. Wang, D. Khabele, S. Orsulic, J. D. Morrow, R. N. DuBois, and S. K. Dey
Extracellular Signal-Regulated Kinase Is a Target of Cyclooxygenase-1-Peroxisome Proliferator-Activated Receptor-{delta} Signaling in Epithelial Ovarian Cancer
Cancer Res., June 1, 2007; 67(11): 5285 - 5292.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Pathol.Home page
W.-L. Yang, K. Q. Cai, J. L. Smedberg, E. R. Smith, A. Klein-Szanto, T. C. Hamilton, and X.-X. Xu
A Reduction of Cyclooxygenase 2 Gene Dosage Counters the Ovarian Morphological Aging and Tumor Phenotype in Wv Mice
Am. J. Pathol., April 1, 2007; 170(4): 1325 - 1336.
[Abstract] [Full Text] [PDF]

Home page
 Hum ReprodHome page
S. Battersby, K.J. Sales, A.R. Williams, R.A. Anderson, S. Gardner, and H.N. Jabbour
Seminal plasma and prostaglandin E2 up-regulate fibroblast growth factor 2 expression in endometrial adenocarcinoma cells via E-series prostanoid-2 receptor-mediated transactivation of the epidermal growth factor receptor and extracellular signal-regulated kinase pathway
Hum. Reprod., January 1, 2007; 22(1): 36 - 44.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
S. Upadhyay, C. Liu, A. Chatterjee, M. O. Hoque, M. S. Kim, J. Engles, W. Westra, B. Trink, E. Ratovitski, and D. Sidransky
LKB1/STK11 Suppresses Cyclooxygenase-2 Induction and Cellular Invasion through PEA3 in Lung Cancer
Cancer Res., August 15, 2006; 66(16): 7870 - 7879.
[Abstract] [Full Text] [PDF]

Home page
 Exp. Biol. Med.Home page
F. Spinella, L. Rosano, S. Decandia, V. Di Castro, A. Albini, G. Elia, P. G. Natali, and A. Bagnato
Antitumor Effect of Green Tea Polyphenol Epigallocatechin-3-Gallate in Ovarian Carcinoma Cells: Evidence for the Endothelin-1 as a Potential Target.
Experimental Biology and Medicine, June 1, 2006; 231(6): 1123 - 1127.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
T. Daikoku, S. Tranguch, I. N. Trofimova, D. M. Dinulescu, T. Jacks, A. Yu. Nikitin, D. C. Connolly, and S. K. Dey
Cyclooxygenase-1 is overexpressed in multiple genetically engineered mouse models of epithelial ovarian cancer.
Cancer Res., March 1, 2006; 66(5): 2527 - 2531.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
V. R. Holla, J. R. Mann, Q. Shi, and R. N. DuBois
Prostaglandin E2 Regulates the Nuclear Receptor NR4A2 in Colorectal Cancer
J. Biol. Chem., February 3, 2006; 281(5): 2676 - 2682.
[Abstract] [Full Text] [PDF]

Home page
 Endocr Relat CancerHome page
A. Bagnato, F. Spinella, and L. Rosano
Emerging role of the endothelin axis in ovarian tumor progression
Endocr. Relat. Cancer, December 1, 2005; 12(4): 761 - 772.
[Abstract] [Full Text] [PDF]

Home page
 J. Thorac. Cardiovasc. Surg.Home page
M. Ates, O. Altundag, S. Aktolga, and K. Altundag
Association between cyclooxygenase and epidermal growth factor receptor pathways in non-small cell lung cancer
J. Thorac. Cardiovasc. Surg., November 1, 2005; 130(5): 1484 - 1485.
[Full Text] [PDF]

Home page
 Cancer Res.Home page
B. H.A. von Rahden, H. J. Stein, F. Puhringer, I. Koch, R. Langer, G. Piontek, J. R. Siewert, H. Hofler, and M. Sarbia
Coexpression of Cyclooxygenases (COX-1, COX-2) and Vascular Endothelial Growth Factors (VEGF-A, VEGF-C) in Esophageal Adenocarcinoma
Cancer Res., June 15, 2005; 65(12): 5038 - 5044.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
T. Daikoku, D. Wang, S. Tranguch, J. D. Morrow, S. Orsulic, R. N. DuBois, and S. K. Dey
Cyclooxygenase-1 Is a Potential Target for Prevention and Treatment of Ovarian Epithelial Cancer
Cancer Res., May 1, 2005; 65(9): 3735 - 3744.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
K. A. Crist, Z. Zhang, M. You, W. T. Gunning, P. B. Conran, V. E. Steele, and R. A. Lubet
Characterization of rat ovarian adenocarcinomas developed in response to direct instillation of 7,12-dimethylbenz[a]anthracene (DMBA) coated suture
Carcinogenesis, May 1, 2005; 26(5): 951 - 957.
[Abstract] [Full Text] [PDF]

Home page
 JCOHome page
J. R. Brown and R. N. DuBois
COX-2: A Molecular Target for Colorectal Cancer Prevention
J. Clin. Oncol., April 20, 2005; 23(12): 2840 - 2855.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
V. R. Holla, D. Wang, J. R. Brown, J. R. Mann, S. Katkuri, and R. N. DuBois
Prostaglandin E2 Regulates the Complement Inhibitor CD55/Decay-accelerating Factor in Colorectal Cancer
J. Biol. Chem., January 7, 2005; 280(1): 476 - 483.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
L. T. Soumaoro, H. Uetake, T. Higuchi, Y. Takagi, M. Enomoto, and K. Sugihara
Cyclooxygenase-2 Expression: A Significant Prognostic Indicator for Patients With Colorectal Cancer
Clin. Cancer Res., December 15, 2004; 10(24): 8465 - 8471.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
F. Spinella, L. Rosano, V. Di Castro, P. G. Natali, and A. Bagnato
Endothelin-1-induced Prostaglandin E2-EP2, EP4 Signaling Regulates Vascular Endothelial Growth Factor Production and Ovarian Carcinoma Cell Invasion
J. Biol. Chem., November 5, 2004; 279(45): 46700 - 46705.
[Abstract] [Full Text] [PDF]

Home page
 Hum Reprod UpdateHome page
J. Sirois, K. Sayasith, K. A. Brown, A. E. Stock, N. Bouchard, and M. Dore
Cyclooxygenase-2 and its role in ovulation: a 2004 account
Hum. Reprod. Update, September 1, 2004; 10(5): 373 - 385.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
F. Spinella, L. Rosano, V. Di Castro, M. R. Nicotra, P. G. Natali, and A. Bagnato
Inhibition of Cyclooxygenase-1 and -2 Expression by Targeting the Endothelin A Receptor in Human Ovarian Carcinoma Cells
Clin. Cancer Res., July 15, 2004; 10(14): 4670 - 4679.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
V. Quidville, N. Segond, E. Pidoux, R. Cohen, A. Jullienne, and S. Lausson
Tumor Growth Inhibition by Indomethacin in a Mouse Model of Human Medullary Thyroid Cancer: Implication of Cyclooxygenases and 15-Hydroxyprostaglandin Dehydrogenase
Endocrinology, May 1, 2004; 145(5): 2561 - 2571.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
H. Wang, W.-g. Ma, L. Tejada, H. Zhang, J. D. Morrow, S. K. Das, and S. K. Dey
Rescue of Female Infertility from the Loss of Cyclooxygenase-2 by Compensatory Up-regulation of Cyclooxygenase-1 Is a Function of Genetic Makeup
J. Biol. Chem., March 12, 2004; 279(11): 10649 - 10658.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
F. G. Bottone Jr, J. M. Martinez, B. Alston-Mills, and T. E. Eling
Gene modulation by Cox-1 and Cox-2 specific inhibitors in human colorectal carcinoma cancer cells
Carcinogenesis, March 1, 2004; 25(3): 349 - 357.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
J. L. Mulshine, J. C. Atkinson, R. O. Greer, V. A. Papadimitrakopoulou, C. Van Waes, S. Rudy, J. W. Martin, S. M. Steinberg, D. J. Liewehr, I. Avis, et al.
Randomized, Double-Blind, Placebo-Controlled Phase IIB Trial of the Cyclooxygenase Inhibitor Ketorolac as an Oral Rinse in Oropharyngeal Leukoplakia
Clin. Cancer Res., March 1, 2004; 10(5): 1565 - 1573.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Cancer Res.Home page
T.-L. Erkinheimo, H. Lassus, P. Finne, B. P. van Rees, A. Leminen, O. Ylikorkala, C. Haglund, R. Butzow, and A. Ristimaki
Elevated Cyclooxygenase-2 Expression Is Associated with Altered Expression of p53 and SMAD4, Amplification of HER-2/neu, and Poor Outcome in Serous Ovarian Carcinoma
Clin. Cancer Res., January 15, 2004; 10(2): 538 - 545.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
J. Raju and R. P. Bird
Energy Restriction Reduces the Number of Advanced Aberrant Crypt Foci and Attenuates the Expression of Colonic Transforming Growth Factor {beta} and Cyclooxygenase Isoforms in Zucker Obese (fa/fa) Rats
Cancer Res., October 15, 2003; 63(20): 6595 - 6601.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
F. G. Buchanan, D. Wang, F. Bargiacchi, and R. N. DuBois
Prostaglandin E2 Regulates Cell Migration via the Intracellular Activation of the Epidermal Growth Factor Receptor
J. Biol. Chem., September 12, 2003; 278(37): 35451 - 35457.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Gupta, R. A.
Right arrow Articles by DuBois, R. N.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Gupta, R. A.
Right arrow Articles by DuBois, R. N.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online