The Effects of Selective Cyclooxygenase-2 Inhibitor and Prostaglandin E2 Receptor Agonists on the Endothelin Axis of Prostate Cancer Cells / 대한비뇨기과학회지

PURPOSE: The enhanced expression of the cyclooxygenase-2 (COX-2), prostaglandin E2 receptor (EPs) and endothelin-1 (ET-1) axis is known to play a significant role in the development and progression of several malignancies. To date, little work has been done to investigate the relationships between the COX-2, EPs and ET-1 axis in prostate cancer (PC) cells. The aim of this study is to investigate the expression of preproET-1 (PPET-1), ET-1 receptor A (ET(A)R), and endothelin converting enzyme-1 (ECE-1) in the PC cell lines and to evaluate the effects of COX-2 and EPs on the expression of PPET-1, ET(A)R, and ECE-1. MATERIALS AND METHODS: Two PC cell lines, PC-3 and DU-145 cells were used for this study. By performing reverse transcription polymerase chain reaction (RT-PCR), the mRNA expressions of PPET-1, ET(A)R and ECE-1 were detected, and then the mRNA expressions of PPET-1, ET(A)R and ECE-1 were detected after being treating the cells with selective COX-2 inhibitor (NS-398), or EP2 (butaprost) and EP4 (misoprostol), which are both agonist of 10(-10), 10(-8) and 10(-6)M. RESULTS: PPET-1, ET(A)R and ECE-1 mRNA were expressed in both cell lines. After NS-398 treatment, only the PPET-1 mRNA expression was decreased at 4, 8 and 12 hours in the PC-3 cells. EP2 and EP4 agonist induced an increase for the PPET-1, ET(A)R and ECE-1 mRNA expressions, compared with the NS-398 treated group (control), in the PC-3 cells. CONCLUSIONS: ET-1/ET(A)R and ECE-1, whose expressions are increased by EP2 and EP4, may play key roles in the development and progression of PC via COX-2. A combination treatment with selective inhibitors for COX-2, EPs and ET(A)R would be novel approach to prostate cancer therapy.

Effects of Granulocyte Macrophage Colony-Stimulating Factor (GM-CSF) on the Interleukin-6 Expression in the Prostate Cancer Cell Line PC-3 / 대한비뇨기과학회지

PURPOSE: Interleukin-6 (IL-6) can stimulate a variety of tumors including prostatic carcinoma. Research has recently shown that IL-6 may act to stimulate the progression of prostatic cancer. IL-6 is elevated in the sera of patients with metastatic prostatic cancer and it has been shown to be a candidate marker of disease activity. To date, little work has been performed to characterize the nature of granulocyte macrophage colony-stimulating factor (GM-CSF) and the expression of IL-6. The aim of this study is to evaluate the effects of GM-CSF on the expression of IL-6 in PC-3 cells. MATERIALS AND METHODS: The bone-derived PC-3 cell line was used in this study. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the GM-CSF and also the IL-6 mRNA expression. The IL-6 protein was measured by enzyme-linked immunosorbent assay (ELISA) after treatments with the hGM-CSF. RESULTS: hGM-CSF was expressed in the PC-3 cell line. Our data indicated that the IL-6 mRNA expression was not increased at 4, 8 and 12 hours by the hGM-CSF in comparison to the control group, but it was slightly increased at 24 and 48 hours. The expression of IL-6 protein was increased at 4, 8, 12, 24 and 48 hours after hGM-CSF treatment, in comparison with the control group. CONCLUSIONS: The IL-6 mRNA expression was slightly increased by hGM-CSF at 24 and 48 hours in comparison to the control group. Yet the IL-6 protein expression increased before the IL-6 mRNA expression. Therefore, hGM-CSF may modulate the post-transcription pathway of the IL-6 expression in prostate carcinoma cells. Our data suggest that GM-CSF may have a possible IL-6 mediated pathophysiologic role in prostate cancer.

Interleukin-1beta Modulates Proliferation, Interleukin-6 and Interleukin Receptor Expression in PC-3 and DU-145 Prostatic Cancer Cells / 대한비뇨기과학회지

Purpose: IL-1 is a multifunctional proinflammatory cytokine. As the proliferative effects of IL-6 and IL-6 receptor expressions on prostatic cancer cells in response to IL-1 have not been determined, the effects of IL-1 on prostatic cancer cell lines were investigated. Materials and Methods: PC-3 and DU-145 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum. Cell cultures were supplemented with various concentrations of IL-1 (0, 1, 10, 20 and 40ng/ml), and the MMT growth assay performed. PC-3 and DU-145 cells were treated for 2, 4, 8, 12 and 24 h both with and without IL-1. IL-6 and IL-6 receptor (IL-6R) mRNA expressions were investigated using RT-PCR, and the IL-6 levels in cultured supernatant measured by ELISA. Results: The viability of both PC-3 and DU-145 cells decreased after IL-1 treatment (10, 20 and 40ng/mul). With 40ng/ml the IL-1, IL-6 and IL-6RmRNA expressions were lower in PC-3 cells, but unchanged in DU-145 cells, whereas the IL-6 protein production was higher in both PC-3 and DU-145 cells. Conclusions: IL-1 inhibited the proliferation of both PC-3 and DU145 cells. In the PC-3 cells, IL-1 decreased the expressions of IL-6 and IL-6R mRNA, but paradoxically increased the IL-6 production. In the DU-145 cells, IL-1 treatment did not affect the IL-6 or IL-6R mRNA expressions, but the IL-6 production was increased. This discrepancy between IL-1-induced IL-6 mRNA and protein production may be mediated by modification to the protein synthesis or an increased cellular excretion.

The Prostaglandin E Receptor Agonists Increase Granulocyte Macrophage Colony-Stimulating Factor in Prostate Cancer Cells / 대한비뇨기과학회지

PURPOSE: The predilection for prostate carcinoma cells to metastasize to bone suggests the hypothesis that bone and/or bone marrow-derived factors may promote prostate carcinoma cell growth and/or their survival. To date, little work has been performed to characterize the nature of granulocyte macrophage colony-stimulating factor (GM-CSF) and the expression of prostaglandin E2 receptors (EPs) in prostate cancer (PC) cells. The aim of this study is to evaluate the effects of GM-CSF on cell proliferation and the effects of EP agonists on the production of GM-CSF in the PC-3 cells. MATERIALS AND METHODS: The bone-derived PC-3 cell line was used in this study. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the mRNA expression of EP1, 2, 3 and 4 and hGM- CSF. 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay was done to estimate the viability of PC-3 cells after hGM-CSF treatment. hGM-CSF was measured by enzyme-linked immunosorbent assay (ELISA) after treatments with the EPs agonist at 10(-10), 10(-8), 10(-6)M, respectively. RESULTS: EP2, 3 and 4 and hGM-CSF were expressed in the PC-3 cell line. Viability of the PC-3 cells was significantly increased by hGM-CSF administration in a dose- and time-dependent manner. Also, our data indicated that EP2, 3 and especially 4 agonists induced a significant dose- dependent increase in hGM-CSF production in comparison to the control group in the conditioned ELISA medium. CONCLUSIONS: These results suggest that GM-CSF may be part of a network of an autocrine-regulatory loop system that modulates the biologic activity of prostate carcinoma cells. Our data suggest that GM-CSF and EPs may represent a possible novel therapeutic target that manipulates the proliferative rate of prostate tumors.

The Prostaglandin E Receptor Agonists Increase Granulocyte Macrophage Colony-Stimulating Factor in Prostate Cancer Cells / 대한비뇨기과학회지

PURPOSE: The predilection for prostate carcinoma cells to metastasize to bone suggests the hypothesis that bone and/or bone marrow-derived factors may promote prostate carcinoma cell growth and/or their survival. To date, little work has been performed to characterize the nature of granulocyte macrophage colony-stimulating factor (GM-CSF) and the expression of prostaglandin E2 receptors (EPs) in prostate cancer (PC) cells. The aim of this study is to evaluate the effects of GM-CSF on cell proliferation and the effects of EP agonists on the production of GM-CSF in the PC-3 cells. MATERIALS AND METHODS: The bone-derived PC-3 cell line was used in this study. Reverse transcription polymerase chain reaction (RT-PCR) was performed to detect the mRNA expression of EP1, 2, 3 and 4 and hGM- CSF. 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay was done to estimate the viability of PC-3 cells after hGM-CSF treatment. hGM-CSF was measured by enzyme-linked immunosorbent assay (ELISA) after treatments with the EPs agonist at 10(-10), 10(-8), 10(-6)M, respectively. RESULTS: EP2, 3 and 4 and hGM-CSF were expressed in the PC-3 cell line. Viability of the PC-3 cells was significantly increased by hGM-CSF administration in a dose- and time-dependent manner. Also, our data indicated that EP2, 3 and especially 4 agonists induced a significant dose- dependent increase in hGM-CSF production in comparison to the control group in the conditioned ELISA medium. CONCLUSIONS: These results suggest that GM-CSF may be part of a network of an autocrine-regulatory loop system that modulates the biologic activity of prostate carcinoma cells. Our data suggest that GM-CSF and EPs may represent a possible novel therapeutic target that manipulates the proliferative rate of prostate tumors.

Prostagladin E Receptor II and IV Increase the Expression of Martrix Metalloproteinase-7 in PC (Prostate Cancer)-3 Cells / 대한비뇨기과학회지

PURPOSE: The effects of PGE2 receptors (EP1, 2, 3, 4) on the proliferation of prostate cancer cells are still unclear. The degradation of the basement membrane by MMP-2, 7, 9 and TIMP-1, 2 is a critical point in tumor invasion and metastasis. We investigated the effects of PGE2 receptors concerning MMP and TIMP after the treatment of COX-2 inhibitors on prostate cancer cell-lines. MATERIALS AND METHODS: Two prostate cancer cell-lines, PC-3 and DU-145 cells were used in this study. RT-PCR were performed to detect the mRNA expression of EP1, 2, 3, 4, MMP-2, 7, 9 and TIMP-1, 2, MMP-7 was measured by ELISA after being treated with the selective EP2 agonist and EP4 agonist 10(-10), 10(-8), 10(-6) microM respectively. RESULTS: EP2, 3 and 4 mRNA were expressed in both cell-lines. After the NS-398 treatment, EP2 and EP4 mRNA expressions decreased in PC-3 cells. While only the MMP-7 mRNA expression decreased in PC-3 cells after NS-398 treatment, after NS-398 with selective EP2 agonist and EP4 agonist, MMP-7 mRNA expression increased. In PC-3 cells, selective EP2 agonist and EP4 agonist induced a significant dose-dependent increase in MMP-7 production in comparison to the NS-398 treatment group (control) in the conditioned ELISA medium. CONCLUSIONS: These results strongly suggest that COX-2, to some extent, contribute to prostate carcinogenesis at the EP2 and EP4 receptor, which could also be explained by increments of MMP-7 in PC-3 cells. Therefore, these findings show that selective EP inhibitor is useful in preventing specific disease progression in prostate cancer.

Induction of CEA-specific Cytotoxic T Lymphocytes by Murine Dendritic Cells Expressing CEA

BACKGROUND: Carcinoembryonic antigen (CEA) is well-known soluble tumor marker frequently detectable in peripheral blood of carcinoma patients and considered as good target for antigen-specific immunotherapy. In this study, we used a replication-deficient adenovirus containing CEA to study CTL induction in vitro after adenovirus-mediated gene transfer into DC. METHODS: DC were obtained from mouse bone marrow and cultured with IL-4 and GM-CSF. For measuring CTL activity, splenocytes were harvested from the mice, which were immunized with DC that had been infected AdV-CEA or pulsed with CEA peptide. Untreated DC was used as a control. Splenocytes were re-stimulated in vitro with DC pulsed with CEA peptide for 7 days and CTL activity with CEA peptide-pulsed EL-4 cells were assessed in a standard 51Cr-release assay. The frequencies of antigen-specific cytokine-secreting T cell were determined with mIFN-gamma ELISPOT. RESULTS: DC infected with recombinant adenovirus expressing CEA induced CEA-specific CTL responses in vivo. Splenocyte induced from mice immunized with AdV-CEA-infected DC increase in the number of IFN-gamma secreting T cells compared with those from mice immunized with CEA peptide-pulsed DC. CONCLUSION: These results suggested that DC infected with recombinant adenovirus has advantages over other forms of vaccination and could provide an alternative approach vaccination therapies.