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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

Effects of overexpression of PGIS and carbaprostacyclin on cell survival. (a) MCF-7 cells were transiently cotransfected with pCDNA3.1COX-2, together with control vector pCDNA3.1, pCDNA3.1mPGIS, and pCDNA3.1mPGISC441A. Cell viability upon exposure to 150 μM sulindac for 24 hours was examined by the MTT assay as compared to vehicle-treated controls (0.1% DMSO). ((b), (c)) Carbaprostacyclin-mediated protection from sulindac sulfone-induced apoptosis. CCRF-CEM cells were treated with 0, 100, and 300 μM sulindac sulfone in the presence of (b) 0.01–1 μM carbaprostacyclin or (c) 0.001–10 mM dbcAMP. 24 hours posttreatment caspase-3 activity was measured by cleavage of fluorogenic substrate Ac-DEVD-AMC.
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fig3: Effects of overexpression of PGIS and carbaprostacyclin on cell survival. (a) MCF-7 cells were transiently cotransfected with pCDNA3.1COX-2, together with control vector pCDNA3.1, pCDNA3.1mPGIS, and pCDNA3.1mPGISC441A. Cell viability upon exposure to 150 μM sulindac for 24 hours was examined by the MTT assay as compared to vehicle-treated controls (0.1% DMSO). ((b), (c)) Carbaprostacyclin-mediated protection from sulindac sulfone-induced apoptosis. CCRF-CEM cells were treated with 0, 100, and 300 μM sulindac sulfone in the presence of (b) 0.01–1 μM carbaprostacyclin or (c) 0.001–10 mM dbcAMP. 24 hours posttreatment caspase-3 activity was measured by cleavage of fluorogenic substrate Ac-DEVD-AMC.

Mentions: Exposure of CCRF-CEM cells to 100 and 300 μM sulindac sulfone induced apoptotic cell death in a dose-dependent manner as analyzed by measurement of caspase-3 activity (Figure 3(b)). Coincubation of CCRF-CEM cells with sulindac sulfone and increasing concentrations of carbaprostacyclin (0.01–1 μM) resulted in a decrease of apoptotic cells by about 50% at either sulindac sulfone concentration and by carbaprostacyclin 1 μM. Thus, treatment with the synthetic PGI2 analogue carbaprostacyclin protected against NSAID-induced apoptosis. Cells were coincubated with sulindac sulfone and the membrane permeable cAMP analogue dibutyryl-cAMP (dbcAMP) to rule out involvement of the classical prostacyclin-receptor IP in protection against apoptotic cell death via elevation of intracellular cAMP. Cell viability was not affected upon treatment with dbcAMP suggesting a mode of action independent of IP modulation (Figure 3(c)).


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)

Effects of overexpression of PGIS and carbaprostacyclin on cell survival. (a) MCF-7 cells were transiently cotransfected with pCDNA3.1COX-2, together with control vector pCDNA3.1, pCDNA3.1mPGIS, and pCDNA3.1mPGISC441A. Cell viability upon exposure to 150 μM sulindac for 24 hours was examined by the MTT assay as compared to vehicle-treated controls (0.1% DMSO). ((b), (c)) Carbaprostacyclin-mediated protection from sulindac sulfone-induced apoptosis. CCRF-CEM cells were treated with 0, 100, and 300 μM sulindac sulfone in the presence of (b) 0.01–1 μM carbaprostacyclin or (c) 0.001–10 mM dbcAMP. 24 hours posttreatment caspase-3 activity was measured by cleavage of fluorogenic substrate Ac-DEVD-AMC.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig3: Effects of overexpression of PGIS and carbaprostacyclin on cell survival. (a) MCF-7 cells were transiently cotransfected with pCDNA3.1COX-2, together with control vector pCDNA3.1, pCDNA3.1mPGIS, and pCDNA3.1mPGISC441A. Cell viability upon exposure to 150 μM sulindac for 24 hours was examined by the MTT assay as compared to vehicle-treated controls (0.1% DMSO). ((b), (c)) Carbaprostacyclin-mediated protection from sulindac sulfone-induced apoptosis. CCRF-CEM cells were treated with 0, 100, and 300 μM sulindac sulfone in the presence of (b) 0.01–1 μM carbaprostacyclin or (c) 0.001–10 mM dbcAMP. 24 hours posttreatment caspase-3 activity was measured by cleavage of fluorogenic substrate Ac-DEVD-AMC.
Mentions: Exposure of CCRF-CEM cells to 100 and 300 μM sulindac sulfone induced apoptotic cell death in a dose-dependent manner as analyzed by measurement of caspase-3 activity (Figure 3(b)). Coincubation of CCRF-CEM cells with sulindac sulfone and increasing concentrations of carbaprostacyclin (0.01–1 μM) resulted in a decrease of apoptotic cells by about 50% at either sulindac sulfone concentration and by carbaprostacyclin 1 μM. Thus, treatment with the synthetic PGI2 analogue carbaprostacyclin protected against NSAID-induced apoptosis. Cells were coincubated with sulindac sulfone and the membrane permeable cAMP analogue dibutyryl-cAMP (dbcAMP) to rule out involvement of the classical prostacyclin-receptor IP in protection against apoptotic cell death via elevation of intracellular cAMP. Cell viability was not affected upon treatment with dbcAMP suggesting a mode of action independent of IP modulation (Figure 3(c)).

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