Invasionsfaktoren uPA/PAI‐1 im Tumorgewebe bei Patientinnen mit primärem Mammakarzinom: Von Forschungsergebnissen zur klinischen Anwendung am Beispiel der NNBC 3-Europe-Studie
“…5 The utilisation of these two markers has been singularly successful in breast cancer, where the two markers are used linearly with definite cut-off points to identify early relapse and poor prognosis. 6,7,[11][12][13][14][15] In view of the fact that the utilisation of uPA and PAI-1 titres as biomarkers for prostate cancer is still in its infancy, parameters which influence the procurement of large data collections, e.g. sample shelf life, storage conditions, extraction efficiency and marker stability will be of great interest.…”
Section: Resultsmentioning
confidence: 99%
“…5 The utilisation of these two markers has been singularly successful in breast cancer, where the two markers are used linearly with definite cut-off points to identify early relapse and poor prognosis. 6,7,11–15 Figure 2.Comparison of uPA/PAI-1 ratios for freeze-dried prostate tissue samples and the corresponding frozen samples from 25 patients presenting with PCa and BPH prostate abnormalities. Horizontal lines represent the mean uPA/PAI-1 ratio in each group of samples.…”
Section: Resultsmentioning
confidence: 99%
“…Determination of the uPA and PAI-1 content was by the FEMTELLE assay (Sekisui Diagnostics, LLC, Lexington, MA, USA), as described elsewhere. 6,7 Total protein was expressed as mg/mL, while the content of uPA and PAI-1 was expressed in ng/mg total protein. To test the effect of freeze-drying on the capacity of the uPA-PAI-1 pair to predict disease state, uPA/PAI-1 ratios were calculated for each patient sample and for each processing technique for further analysis.…”
Background: Urokinase plasminogen activator (uPA) and its inhibitor (PAI-1) have been shown to be of merit as biomarkers for a variety of cancers. Prostate tissue resections from patients with prostate cancer (PCa) and benign prostatic hyperplasia were analysed to determine the influence of freeze-drying on the recovery of uPA and PAI-1 and their predictive performance. Methods: Prostate tissue was frozen in liquid nitrogen and homogenised into a fine powder in a precooled stainless steel punch homogeniser. One aliquot of the powder was extracted directly, and a second aliquot was freeze-dried overnight and then extracted. The extracts were analysed by FEMTELLE assay to determine the concentrations of uPA and PAI-1. uPA/PAI-1 ratios were calculated for each sample, and the mean ratios for the frozen and the lyophilised tissue were compared. Results: The concentrations of uPA measured for the frozen and lyophilised samples are strongly correlated (R ¼ 0.90 AE 0.05). The same applies to the PAI-1 measured (R ¼ 0.89 AE 0.03). The uPA/PAI-1 ratios for the lyophilised and frozen samples were strongly correlated. The uPA/PAI-1 ratios for frozen and lyophilised samples were found to be essentially the same with values of 0.0344 AE 0.0066 and 0.0340 AE 0.0068, respectively (P ¼ 0.9633). Conclusion: The recovery of uPA and PAI-1 from a deep frozen prostate tissue homogenate followed by freeze-drying proceeds with a loss of 10 and 11%, respectively, with no influence on the uPA/PAI-1 ratio. Lyophilisation is a safe procedure for the preservation of frozen prostate tissue samples as it permits recovery of the markers without compromising their use for diagnostic purposes.
“…5 The utilisation of these two markers has been singularly successful in breast cancer, where the two markers are used linearly with definite cut-off points to identify early relapse and poor prognosis. 6,7,[11][12][13][14][15] In view of the fact that the utilisation of uPA and PAI-1 titres as biomarkers for prostate cancer is still in its infancy, parameters which influence the procurement of large data collections, e.g. sample shelf life, storage conditions, extraction efficiency and marker stability will be of great interest.…”
Section: Resultsmentioning
confidence: 99%
“…5 The utilisation of these two markers has been singularly successful in breast cancer, where the two markers are used linearly with definite cut-off points to identify early relapse and poor prognosis. 6,7,11–15 Figure 2.Comparison of uPA/PAI-1 ratios for freeze-dried prostate tissue samples and the corresponding frozen samples from 25 patients presenting with PCa and BPH prostate abnormalities. Horizontal lines represent the mean uPA/PAI-1 ratio in each group of samples.…”
Section: Resultsmentioning
confidence: 99%
“…Determination of the uPA and PAI-1 content was by the FEMTELLE assay (Sekisui Diagnostics, LLC, Lexington, MA, USA), as described elsewhere. 6,7 Total protein was expressed as mg/mL, while the content of uPA and PAI-1 was expressed in ng/mg total protein. To test the effect of freeze-drying on the capacity of the uPA-PAI-1 pair to predict disease state, uPA/PAI-1 ratios were calculated for each patient sample and for each processing technique for further analysis.…”
Background: Urokinase plasminogen activator (uPA) and its inhibitor (PAI-1) have been shown to be of merit as biomarkers for a variety of cancers. Prostate tissue resections from patients with prostate cancer (PCa) and benign prostatic hyperplasia were analysed to determine the influence of freeze-drying on the recovery of uPA and PAI-1 and their predictive performance. Methods: Prostate tissue was frozen in liquid nitrogen and homogenised into a fine powder in a precooled stainless steel punch homogeniser. One aliquot of the powder was extracted directly, and a second aliquot was freeze-dried overnight and then extracted. The extracts were analysed by FEMTELLE assay to determine the concentrations of uPA and PAI-1. uPA/PAI-1 ratios were calculated for each sample, and the mean ratios for the frozen and the lyophilised tissue were compared. Results: The concentrations of uPA measured for the frozen and lyophilised samples are strongly correlated (R ¼ 0.90 AE 0.05). The same applies to the PAI-1 measured (R ¼ 0.89 AE 0.03). The uPA/PAI-1 ratios for the lyophilised and frozen samples were strongly correlated. The uPA/PAI-1 ratios for frozen and lyophilised samples were found to be essentially the same with values of 0.0344 AE 0.0066 and 0.0340 AE 0.0068, respectively (P ¼ 0.9633). Conclusion: The recovery of uPA and PAI-1 from a deep frozen prostate tissue homogenate followed by freeze-drying proceeds with a loss of 10 and 11%, respectively, with no influence on the uPA/PAI-1 ratio. Lyophilisation is a safe procedure for the preservation of frozen prostate tissue samples as it permits recovery of the markers without compromising their use for diagnostic purposes.
“…The Amsterdam gene signature is currently validated in a prospective and randomized fashion versus the common clinical-pathological criteria in the European study MINDACT [9]. An alternative approach is evaluated in the ongoing study NNBC 3-Europe which aims at validating the urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 for clinical routine risk assessment and tailoring adjuvant chemotherapy in node-negative high-risk patients [10]. uPA and PAI are Thomssen es was recently identified as a predictive marker of outcome by Mortimer et al [16]: In their study with nearly 900 patients and a follow-up of more than 7 years, the recurrence-free survival was stronger related to the incidence of hot flashes than to age, hormone receptor status, and tumor stage.…”
Section: Defining Risk Of Recurrence In the Individual Patientmentioning
Using a contingent of 62 patients of which 46 were BPH and 16 were PCa, we report definitive concentrations of uPA and PAI-1 in tumour tissue extracts and show that the uPA/PAI-1 ratio emerges as a candidate marker to distinguish between BPH and PCa.
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