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Expression of Prostacyclin-Synthase in Human Breast Cancer: Negative Prognostic Factor and Protection against Cell Death In Vitro.

Klein T, Benders J, Roth F, Baudler M, Siegle I, Kömhoff M - Mediators Inflamm. (2015)

Bottom Line: Endogenously formed prostacyclin (PGI2) and synthetic PGI2 analogues have recently been shown to regulate cell survival in various cell lines.PGIS expression was observed in tumor cells in 48.7% of samples and was associated with a statistically significant reduction in 10-year survival (P = 0.038; n = 193).Transient transfection of PGIS into MCF-7 cells exposed to sulindac increased cell viability by 50% and exposure to carbaprostacyclin protected against sulindac sulfone induced apoptosis in CCRF-CEM cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Philipps University, 35033 Marburg, Germany.

ABSTRACT
Endogenously formed prostacyclin (PGI2) and synthetic PGI2 analogues have recently been shown to regulate cell survival in various cell lines. To elucidate the significance of PGI2 in human breast cancer, we performed immunohistochemistry to analyze expression of prostacyclin-synthase (PGIS) in 248 human breast cancer specimens obtained from surgical pathology files. We examined patients' 10-year survival retrospectively by sending a questionnaire to their general practitioners and performed univariate analysis to determine whether PGIS expression correlated with patient survival. Lastly, the effects of PGI2 and its analogues on cell death were examined in a human breast cancer cell line (MCF-7) and a human T-cell leukemia cell line (CCRF-CEM). PGIS expression was observed in tumor cells in 48.7% of samples and was associated with a statistically significant reduction in 10-year survival (P = 0.038; n = 193). Transient transfection of PGIS into MCF-7 cells exposed to sulindac increased cell viability by 50% and exposure to carbaprostacyclin protected against sulindac sulfone induced apoptosis in CCRF-CEM cells. Expression of PGIS is correlated with a reduced patient survival and protects against cell death in vitro, suggesting that PGIS is a potential therapeutic target in breast cancer.

No MeSH data available.


Related in: MedlinePlus

Immunological detection of prostacyclin-synthase. (a) Western blot analysis of MCF-7 cells transfected with control vector (lane I: pCDNA 3.1), wild-type (lane II: pCDNA3.1mPGIS), and mutant prostacyclin-synthase (lane III; pCDNA3.1 mPGISC441A). As positive control PGIS from bovine aorta is shown on the left. ((b), (c)) Expression of immunoreactive PGIS in tumor cells of a ductal carcinoma showing moderate (b) or intense labeling (c). Slides were photographed at 63x magnification.
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fig1: Immunological detection of prostacyclin-synthase. (a) Western blot analysis of MCF-7 cells transfected with control vector (lane I: pCDNA 3.1), wild-type (lane II: pCDNA3.1mPGIS), and mutant prostacyclin-synthase (lane III; pCDNA3.1 mPGISC441A). As positive control PGIS from bovine aorta is shown on the left. ((b), (c)) Expression of immunoreactive PGIS in tumor cells of a ductal carcinoma showing moderate (b) or intense labeling (c). Slides were photographed at 63x magnification.

Mentions: To confirm expression of wild-type and mutant PGIS protein, Western blot analysis was performed using MCF-7 cell lysates (20 μg per lane). In Figure 1(a), a band could not be detected in cells transfected with the control vector pCDNA 3.1 (lane I). Bands of approximately 52 kD corresponding to the molecular weight of PGIS could be detected in cells transfected with both the PGIS wild-type vector pCDNA3.1mPGIS (lane II) and the mutant PGIS vector pCDNA3.1 mPGISC441A (lane III). Purified PGIS from bovine aorta endothelial cell (left lane) served as positive control.


Expression of Prostacyclin-Synthase in Human Breast Cancer: Negative Prognostic Factor and Protection against Cell Death In Vitro.

Klein T, Benders J, Roth F, Baudler M, Siegle I, Kömhoff M - Mediators Inflamm. (2015)

Immunological detection of prostacyclin-synthase. (a) Western blot analysis of MCF-7 cells transfected with control vector (lane I: pCDNA 3.1), wild-type (lane II: pCDNA3.1mPGIS), and mutant prostacyclin-synthase (lane III; pCDNA3.1 mPGISC441A). As positive control PGIS from bovine aorta is shown on the left. ((b), (c)) Expression of immunoreactive PGIS in tumor cells of a ductal carcinoma showing moderate (b) or intense labeling (c). Slides were photographed at 63x magnification.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4526217&req=5

fig1: Immunological detection of prostacyclin-synthase. (a) Western blot analysis of MCF-7 cells transfected with control vector (lane I: pCDNA 3.1), wild-type (lane II: pCDNA3.1mPGIS), and mutant prostacyclin-synthase (lane III; pCDNA3.1 mPGISC441A). As positive control PGIS from bovine aorta is shown on the left. ((b), (c)) Expression of immunoreactive PGIS in tumor cells of a ductal carcinoma showing moderate (b) or intense labeling (c). Slides were photographed at 63x magnification.
Mentions: To confirm expression of wild-type and mutant PGIS protein, Western blot analysis was performed using MCF-7 cell lysates (20 μg per lane). In Figure 1(a), a band could not be detected in cells transfected with the control vector pCDNA 3.1 (lane I). Bands of approximately 52 kD corresponding to the molecular weight of PGIS could be detected in cells transfected with both the PGIS wild-type vector pCDNA3.1mPGIS (lane II) and the mutant PGIS vector pCDNA3.1 mPGISC441A (lane III). Purified PGIS from bovine aorta endothelial cell (left lane) served as positive control.

Bottom Line: Endogenously formed prostacyclin (PGI2) and synthetic PGI2 analogues have recently been shown to regulate cell survival in various cell lines.PGIS expression was observed in tumor cells in 48.7% of samples and was associated with a statistically significant reduction in 10-year survival (P = 0.038; n = 193).Transient transfection of PGIS into MCF-7 cells exposed to sulindac increased cell viability by 50% and exposure to carbaprostacyclin protected against sulindac sulfone induced apoptosis in CCRF-CEM cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Philipps University, 35033 Marburg, Germany.

ABSTRACT
Endogenously formed prostacyclin (PGI2) and synthetic PGI2 analogues have recently been shown to regulate cell survival in various cell lines. To elucidate the significance of PGI2 in human breast cancer, we performed immunohistochemistry to analyze expression of prostacyclin-synthase (PGIS) in 248 human breast cancer specimens obtained from surgical pathology files. We examined patients' 10-year survival retrospectively by sending a questionnaire to their general practitioners and performed univariate analysis to determine whether PGIS expression correlated with patient survival. Lastly, the effects of PGI2 and its analogues on cell death were examined in a human breast cancer cell line (MCF-7) and a human T-cell leukemia cell line (CCRF-CEM). PGIS expression was observed in tumor cells in 48.7% of samples and was associated with a statistically significant reduction in 10-year survival (P = 0.038; n = 193). Transient transfection of PGIS into MCF-7 cells exposed to sulindac increased cell viability by 50% and exposure to carbaprostacyclin protected against sulindac sulfone induced apoptosis in CCRF-CEM cells. Expression of PGIS is correlated with a reduced patient survival and protects against cell death in vitro, suggesting that PGIS is a potential therapeutic target in breast cancer.

No MeSH data available.


Related in: MedlinePlus