- Email: [email protected]
COX and friends
Available online at www.sciencedirect.com
SCIENCE ~ O l R E C T ® ELSEVIER
BIOMEDICINE PHARMACOTHERAI~
Biomedicine & Pharmacotherapy59 (2005) $317-$320 http://france.elsevier.com/direct/BIOPHA/ COX-2
Abstracts Correlation between Cox-2 related gene expressions and response to preoperative chemotherapy with celecoxib in primary breast cancer H. Bando a, ,, L.W.C. Chow b, M. Higashimoto c, W.T. Loo, C. Nakanishi a, y. Takebayashi, M. Toi a a Breast Cancer Research Group, Tokyo Metropolitan Cancer and Infectious Diseases Center, Honkomagome, 3-1822, Bunkyo-ku, 113-0087 Tokyo, Japan b Department of Surgery, The University of Hong Kong, Hong Kong c Department of Surgery II, Fukushima Medical University, Hikarigaoka 1, 960-1295 Fukushima, Japan
* Corresponding author. (H. Bando) Inhibition of cyclooxygenase (COX)-2 enhances antitumor activity of chemotherapy in the preclinical experiment. According to this hypothesis, we have conducted a pilot study with FEC (cyclophosphamide, epirubicin and 5-FU) plus celecoxib (800 mg/day) in preoperative setting for primary breast cancer. In this study, we aimed to investigate the correlation between clinical and pathological responses and expressions of COX-2 related genes. Tumor samples were collected from twenty-three primary breast cancer patients before and after the treatment and the extracted mRNA was
used for real-time PCR analysis. Twenty-five genes related to COX-2, COX-l, hypoxia and angiogenesis were chosen for analysis. Several gene expressions at pre-treatment and the changes of gene expression by the treatment were significantly correlated with anti-tumor activity of anthracyclinecontaining chemotherapy combined with COX-2 inhibition. COX and friends Y. Takebayashi
Department of Surgery, Institute of Biomedical Sciences Fukushima Medical, University School of Medicine, Fukushima, Japan Considerable reports in vitro, animal model, and epidemiologic evidence suggest that COX-2 may play some role in several types of human solid tumors. In addition, COX-2 inhibitors could be one of a promising drug for cancer chemotherapy. However, the mechanism(s) of COX-2 inhibitor and regulation of COX-2 by anti-cancer drug(s) are still not clear in vitro and in vivo. To resolve these questions, we performed several experiments. I wish the following issues with our data will be discussed. • Relationship(s) between COX-2 and COX- 1, arachidonic acid, phospholipase and PPAR alpha and gamma in anti-cancer drug treated cells (see Fig. 1).
Phospholipase activating protein(PLAA)
l PLA2 2onfer to release AA from :ellular membrane. PLA2:PhospholipaseA2
COX1,2 LOX: Lipoxygenase PG: Prostaglandin TXA2: Thromboxanes HPETE: Hydroperoxyeicosatetraenoic acid HETE: Hydroxyeicosatetraenoic acid LT: Leukotrien
Fig. 1 Relationship(s)between COX-2,COX-l, arachidonic acid, phospholipaseand PPAR alpha and gammain anti-cancerdrug treated cells. © 2005 ElsevierSAS. All rights reserved.
$318
Abstracts/ Biomedicine &Pharmacotherapy59 (2005)$317-$320
• Modified quantitative PCR method for chemotherapy in clinic. • Expression and mutation(s) of COX-2 and COX-l, arachidonic acid, phospholipase and PPAR alpha and gamma in clinical samples, and their significance in clinic.
Cyclooxygenase-2: possible predictor related to micrometastasis in breast cancer patients L. Zhu, W.T.Y. Loo, L.W.C. Chow * Hung Chao Hong Integrated Center for Breast Diseases, Department of Surgery, The University of Hong Kong Medical Center, Pokfulam Road, Pokfulam, Hong Kong • Corresponding author. (L. W.C. Chow) B a c k g r o u n d . - Although micro-metastasis has been successfully hunted by variety of detection methods, little is known about the relationship between micro-metastasis and primary tumors. Cyclooxygenase-2 (COX-2) overexpression has been found to be associated with lymph node metastasis. The purpose of this study was to assess the association of COX-2 expression with micro-metastasis in breast cancer as well as to inquire about its significance as predictor for micro-metastasis. M e t h o d s . - As destination sites for micro-metastasis, we examined peripheral blood, bone marrow (BM) and sentinel lymph node (SLN) specimens from 53 breast cancer patients. Both blood and bone marrow samples were handled using magnetic-activated cell separation (MACS) followed by immunocytochemistry (ICC). The expressions of COX-2 at the primary site were both determined by immunohistochemistry (IHC) and real-time RT-PCR. Results. - The percentages of the patients with micrometastasis were 24.5% for peripheral blood, 56.6% for BM. SLN micro-metastasis were detected in 26.4% and 41.5% patients using H and E and ICC. The amplification of COX2 within primary tumors was significantly associated with SLN micro-metastasis (P = 0.03). And the overexpression of COX-2 was significantly associated with the micrometastasis both in BM (P = 0.005) and SLN (P < 0.001). C o n c l u s i o n s . - Our results indicate possible value of COX-2 to predict the risk of micro-metastasis in breast cancers. Moreover, these results raise the possibility that selective COX-2 inhibitors may be effective in the prevention and treatment of micro-metastasis in breast cancer patients.
Down syndrome critical region proteinl (DSCR1), a novel VEGF target gene that regulates expression of COX-2 on activated endothelial cells B.A. Hesser, X.H. Liang, G. Camenisch a, S. Yang, D. Lewin b, R. Scheller, N. Ferrara, H.-P. Gerber * Department of Molecular Oncology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA a Present address: Institute of Physiology, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Germany
Present address: CuraGen Corporation, New Haven, CT 06511, USA.
b
We conducted a genome wide analysis of genes that are regulated by VEGF in endothelial cells and identified DSCR1 to be most significantly induced. Consistent with an antagonistic function on calcineurin (CnA) signaling, expression of DSCR1 in endothelial cells blocked dephosphorylation, nuclear translocation and activity of NFAT, a transcription factor involved in mediating CnA signaling. DSCR1 was not only induced by VEGF, but also by other compounds activating CnA signaling, suggesting a more general role for DSCR 1 in activated endothelial cells. Transient expression of DSCR 1 attenuated inflammatory marker genes such as tissue factor (TF), E-selectin and COX-2, identifying a previously unknown regulatory role for DSCR1 in activated endothelial cells. In contrast, knock-down of endogenous DSCR1 increased NFAT activity and stimulated expression of inflammatory genes on activated endothelial cells. Thus, the negative regulatory feed back loop between DSCR1 and CnA signaling in endothelial cells identified may represent a potential molecular mechanism underlying the frequently transient expression of inflammatory genes following activation of endothelial cells.
Inhibition effect of human angiogenesis of the cyclooxygenase (COX)-2 inhibitor with MMT cells S.-Y. Kim, T.-Y. Kim
Department of Surgery, College of Medicine, Soonchunhyang University, Cheonan, Chungnam, South Korea Angiogenesis plays a key role in the growth and metastasis tumor. Angio is reportedly enhanced by PGs. COX-2, an inducible enzyme, which catalyzes the formation of PGs form arachidonic acid. We performed our study to determine that the effect of COX-2 inhibitor on experiment with mouse mammary tumor (MMT) cell line in vivo. C57BL/6 mice were randomize to receive 35 days of either placebo (n = 11) or 1500 ppm celecoxib-supplement diet (n = 13). Animals were sacrificed 21 days later and skin specimens were harvested/ processed for quantification of microvessel density associated with each inoculated site. Aortas were used to create identical aortic ring angiogenesis assay. According to immunohistochemical stain, celecoxib administration resulted in parallel decreased on MVD at both control and MMT inoculated sites (22% and 21%, retrospectively, P = 0.025, P = 0.010). In aortic ring assay, the initiation of dietary treatment group was not significantly inhibited to compare with placebo group (75% and 63.3%, retrospectively, P = NS). But dietary celecoxib administration significantly inhibited angiogenetic index of neovessel growth rate. These results suggest that a COX-2 inhibitor has an antiangiogenetic effect in the mammary cancer cell in vivo. When we perform more investigation of a COX-2 inhibitor, it will be crucial drugs as new chemotherapy agent in breast cancer.
SCIENCE ~ O l R E C T ® ELSEVIER
BIOMEDICINE PHARMACOTHERAI~
Biomedicine & Pharmacotherapy59 (2005) $317-$320 http://france.elsevier.com/direct/BIOPHA/ COX-2
Abstracts Correlation between Cox-2 related gene expressions and response to preoperative chemotherapy with celecoxib in primary breast cancer H. Bando a, ,, L.W.C. Chow b, M. Higashimoto c, W.T. Loo, C. Nakanishi a, y. Takebayashi, M. Toi a a Breast Cancer Research Group, Tokyo Metropolitan Cancer and Infectious Diseases Center, Honkomagome, 3-1822, Bunkyo-ku, 113-0087 Tokyo, Japan b Department of Surgery, The University of Hong Kong, Hong Kong c Department of Surgery II, Fukushima Medical University, Hikarigaoka 1, 960-1295 Fukushima, Japan
* Corresponding author. (H. Bando) Inhibition of cyclooxygenase (COX)-2 enhances antitumor activity of chemotherapy in the preclinical experiment. According to this hypothesis, we have conducted a pilot study with FEC (cyclophosphamide, epirubicin and 5-FU) plus celecoxib (800 mg/day) in preoperative setting for primary breast cancer. In this study, we aimed to investigate the correlation between clinical and pathological responses and expressions of COX-2 related genes. Tumor samples were collected from twenty-three primary breast cancer patients before and after the treatment and the extracted mRNA was
used for real-time PCR analysis. Twenty-five genes related to COX-2, COX-l, hypoxia and angiogenesis were chosen for analysis. Several gene expressions at pre-treatment and the changes of gene expression by the treatment were significantly correlated with anti-tumor activity of anthracyclinecontaining chemotherapy combined with COX-2 inhibition. COX and friends Y. Takebayashi
Department of Surgery, Institute of Biomedical Sciences Fukushima Medical, University School of Medicine, Fukushima, Japan Considerable reports in vitro, animal model, and epidemiologic evidence suggest that COX-2 may play some role in several types of human solid tumors. In addition, COX-2 inhibitors could be one of a promising drug for cancer chemotherapy. However, the mechanism(s) of COX-2 inhibitor and regulation of COX-2 by anti-cancer drug(s) are still not clear in vitro and in vivo. To resolve these questions, we performed several experiments. I wish the following issues with our data will be discussed. • Relationship(s) between COX-2 and COX- 1, arachidonic acid, phospholipase and PPAR alpha and gamma in anti-cancer drug treated cells (see Fig. 1).
Phospholipase activating protein(PLAA)
l PLA2 2onfer to release AA from :ellular membrane. PLA2:PhospholipaseA2
COX1,2 LOX: Lipoxygenase PG: Prostaglandin TXA2: Thromboxanes HPETE: Hydroperoxyeicosatetraenoic acid HETE: Hydroxyeicosatetraenoic acid LT: Leukotrien
Fig. 1 Relationship(s)between COX-2,COX-l, arachidonic acid, phospholipaseand PPAR alpha and gammain anti-cancerdrug treated cells. © 2005 ElsevierSAS. All rights reserved.
$318
Abstracts/ Biomedicine &Pharmacotherapy59 (2005)$317-$320
• Modified quantitative PCR method for chemotherapy in clinic. • Expression and mutation(s) of COX-2 and COX-l, arachidonic acid, phospholipase and PPAR alpha and gamma in clinical samples, and their significance in clinic.
Cyclooxygenase-2: possible predictor related to micrometastasis in breast cancer patients L. Zhu, W.T.Y. Loo, L.W.C. Chow * Hung Chao Hong Integrated Center for Breast Diseases, Department of Surgery, The University of Hong Kong Medical Center, Pokfulam Road, Pokfulam, Hong Kong • Corresponding author. (L. W.C. Chow) B a c k g r o u n d . - Although micro-metastasis has been successfully hunted by variety of detection methods, little is known about the relationship between micro-metastasis and primary tumors. Cyclooxygenase-2 (COX-2) overexpression has been found to be associated with lymph node metastasis. The purpose of this study was to assess the association of COX-2 expression with micro-metastasis in breast cancer as well as to inquire about its significance as predictor for micro-metastasis. M e t h o d s . - As destination sites for micro-metastasis, we examined peripheral blood, bone marrow (BM) and sentinel lymph node (SLN) specimens from 53 breast cancer patients. Both blood and bone marrow samples were handled using magnetic-activated cell separation (MACS) followed by immunocytochemistry (ICC). The expressions of COX-2 at the primary site were both determined by immunohistochemistry (IHC) and real-time RT-PCR. Results. - The percentages of the patients with micrometastasis were 24.5% for peripheral blood, 56.6% for BM. SLN micro-metastasis were detected in 26.4% and 41.5% patients using H and E and ICC. The amplification of COX2 within primary tumors was significantly associated with SLN micro-metastasis (P = 0.03). And the overexpression of COX-2 was significantly associated with the micrometastasis both in BM (P = 0.005) and SLN (P < 0.001). C o n c l u s i o n s . - Our results indicate possible value of COX-2 to predict the risk of micro-metastasis in breast cancers. Moreover, these results raise the possibility that selective COX-2 inhibitors may be effective in the prevention and treatment of micro-metastasis in breast cancer patients.
Down syndrome critical region proteinl (DSCR1), a novel VEGF target gene that regulates expression of COX-2 on activated endothelial cells B.A. Hesser, X.H. Liang, G. Camenisch a, S. Yang, D. Lewin b, R. Scheller, N. Ferrara, H.-P. Gerber * Department of Molecular Oncology, Genentech, Inc., 1 DNA Way, South San Francisco, CA 94080, USA a Present address: Institute of Physiology, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Germany
Present address: CuraGen Corporation, New Haven, CT 06511, USA.
b
We conducted a genome wide analysis of genes that are regulated by VEGF in endothelial cells and identified DSCR1 to be most significantly induced. Consistent with an antagonistic function on calcineurin (CnA) signaling, expression of DSCR1 in endothelial cells blocked dephosphorylation, nuclear translocation and activity of NFAT, a transcription factor involved in mediating CnA signaling. DSCR1 was not only induced by VEGF, but also by other compounds activating CnA signaling, suggesting a more general role for DSCR 1 in activated endothelial cells. Transient expression of DSCR 1 attenuated inflammatory marker genes such as tissue factor (TF), E-selectin and COX-2, identifying a previously unknown regulatory role for DSCR1 in activated endothelial cells. In contrast, knock-down of endogenous DSCR1 increased NFAT activity and stimulated expression of inflammatory genes on activated endothelial cells. Thus, the negative regulatory feed back loop between DSCR1 and CnA signaling in endothelial cells identified may represent a potential molecular mechanism underlying the frequently transient expression of inflammatory genes following activation of endothelial cells.
Inhibition effect of human angiogenesis of the cyclooxygenase (COX)-2 inhibitor with MMT cells S.-Y. Kim, T.-Y. Kim
Department of Surgery, College of Medicine, Soonchunhyang University, Cheonan, Chungnam, South Korea Angiogenesis plays a key role in the growth and metastasis tumor. Angio is reportedly enhanced by PGs. COX-2, an inducible enzyme, which catalyzes the formation of PGs form arachidonic acid. We performed our study to determine that the effect of COX-2 inhibitor on experiment with mouse mammary tumor (MMT) cell line in vivo. C57BL/6 mice were randomize to receive 35 days of either placebo (n = 11) or 1500 ppm celecoxib-supplement diet (n = 13). Animals were sacrificed 21 days later and skin specimens were harvested/ processed for quantification of microvessel density associated with each inoculated site. Aortas were used to create identical aortic ring angiogenesis assay. According to immunohistochemical stain, celecoxib administration resulted in parallel decreased on MVD at both control and MMT inoculated sites (22% and 21%, retrospectively, P = 0.025, P = 0.010). In aortic ring assay, the initiation of dietary treatment group was not significantly inhibited to compare with placebo group (75% and 63.3%, retrospectively, P = NS). But dietary celecoxib administration significantly inhibited angiogenetic index of neovessel growth rate. These results suggest that a COX-2 inhibitor has an antiangiogenetic effect in the mammary cancer cell in vivo. When we perform more investigation of a COX-2 inhibitor, it will be crucial drugs as new chemotherapy agent in breast cancer.