Comparison of immunohistochemistry with immunoassay (ELISA) for the detection of components of the plasminogen activation system in human tumour tissue

Ferrier, C.M.; Witte, H H de; Straatman, Huub; Tienoven, Doorlène H. van; Geloof, W L van; Rietveld, F.J.R.; Sweep, Fred C.G.J.; Ruiter, Dirk J.; Muijen, G.N.P. van

doi:10.1038/sj.bjc.6690245

Cited by 41 publications

(30 citation statements)

References 24 publications

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“…This tissue heterogeneity may be one reason for the discrepancy between immunohistochemistry and ELISA results in the present as well as in former studies [6]. In our cases, a correspondence was only seen in tumors lacking stromal uPA expression which also showed negative ELISA results.…”

Section: Discussioncontrasting

confidence: 70%

“…Both enzymes are not only expressed by the cancer cells themselves pointing to their function in cancer cell migration and invasion [1,4]. They are also expressed by fibroblast-like stromal cells, macrophages, mastcells and endothelial cells [2,4,6,12]. Secretion of matrix-degrading proteases, as well as their downstream activators such as uPA by fibroblasts of the tumor stroma underlines their critical role for tumor growth [3,10,13,15].…”

Section: Discussionmentioning

confidence: 99%

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Influence of preoperative core biopsies on uPA/PAI-1 expression in breast cancer tissue

Haas¹,

Park

Hahne³

et al. 2008

Virchows Arch

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The urokinase-type plasminogen activator (uPA) and the plasminogen activator inhibitor-1 (PAI-1) are involved in tumor invasion and metastasis as well as wound healing and inflammation. We investigated the impact of tissue injury by preoperative sampling on the level of uPA/PAI-1 in paraffin tissue of 55 breast cancer cases by immunohistochemistry. The tumor area surrounding the biopsy channel was compared with distant intact tumor tissue. uPA and PAI-1 were constantly expressed by the tumor cells. Fibroblastic expression was higher in the tumor area surrounding the biopsy channel than in intact tissue with 47 vs 29 positive cases for uPA (p<0.001) and 35 vs 25 positive cases for PAI-1 (p=0.055). A decrease in fibroblastic enzyme expression from the biopsy area to the intact tumor tissue was seen in 21 cases for uPA and 16 cases for PAI-1. This difference was most evident in cases with an interval of 6 days and longer between biopsy and surgery. In conclusion, fibroblastic inflammatory reaction around the biopsy channel affects stromal uPA and PAI-1 expression, which possibly leads to falsely increased enzyme levels in ELISA. Tissue specimen for ELISA analysis should not be taken from the tumor area around the biopsy channel in the resection specimen.

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Section: Discussioncontrasting

confidence: 70%

Section: Discussionmentioning

confidence: 99%

Influence of preoperative core biopsies on uPA/PAI-1 expression in breast cancer tissue

Haas¹,

Park

Hahne³

et al. 2008

Virchows Arch

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“…However, it has been surprisingly difficult to reach a consensus on the localization of PAI-1 because PAI-1 immunoreactivity was reported to be present solely or mainly in cancer cells by some authors, 29,30 mainly in the stromal fibroblast-like cells by others, 31 or both in cancer cells and stromal fibroblast-like cells by still others. [32][33][34][35][36][37] The difficulty in reaching conclusive results should be seen in relation to the low amounts of PAI-1 in breast tumors, in extracts in the order of magnitude of 10 ng/mg total protein. This fact renders the requirement for strict specificity controls particularly pertinent.…”

Section: Discussionmentioning

confidence: 99%

“…46 In brief, 3-m paraffin sections were deparaffinized in xylene, hydrated through graded ethanol solutions, and digested with protease K (5 g/ml) for 5 minutes at 44°C. Sections were then dehydrated and the 35 S-labeled probes (2 ϫ 10 6 cpm/slide) incubated overnight at 55°C in a humidified chamber. Sections were washed with agitation at 55°C with a buffer of 15 mmol/L sodium citrate, pH 7.0, 0.15 mol/L NaCl (SSC) containing 0.1% SDS and 10 mmol/L dithiothreitol for 10 minutes in 2ϫ SSC, for 10 minutes in 0.5ϫ SSC, and for 10 minutes in 0.2ϫ SSC.…”

Section: In Situ Hybridizationmentioning

confidence: 99%

The Myofibroblast Is the Predominant Plasminogen Activator Inhibitor-1-Expressing Cell Type in Human Breast Carcinomas

Offersen

Nielsen²,

Høyer‐Hansen³

et al. 2003

The American Journal of Pathology

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The tumor level of plasminogen activator inhibitor-1 (PAI-1) is an informative biochemical marker of a poor prognosis in several cancer types. However, the tumor biological functions of PAI-1 and the identity of PAI-1-expressing cells are controversial. With the aim of immunohistochemically localizing PAI-1 in formalin-fixed, paraffin-embedded invasive ductal breast carcinoma samples, we raised new polyclonal antibodies against PAI-1 from different expression systems. The antibodies were affinity purified by absorption on immobilized preparations of PAI-1 different from those used for immunization. The specificity of the antibodies was ensured by immunoblotting analysis. In immunohistochemistry, the staining pattern obtained with the antibodies showed a good correlation with the PAI-1 mRNA expression pattern. In all 25 cases analyzed, PAI-1 immunoreactivity was predominantly localized in fibroblast-like cells. Double-immunofluorescence analyses showed co-expression of PAI-1 and ␣-smooth muscle actin in these cells, suggesting that they are myofibroblasts. PAI-1 was also seen in some myoepithelial cells surrounding occasional foci of ductal carcinoma in situ (9 of 25), some endothelial cells (8 of 25), some cancer cells (3 of 25), and some mast cells (6 of 25). In conclusion, we have provided a robust immunohistochemical procedure for detection of PAI-1 and shown that the majority of the PAI-1-expressing cells in invasive ductal breast carcinomas are myofibroblasts.

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“…Using this system, antigens can be detected easily without a washing procedure such as those required in immunochromatographic assays. (8) However, the detection limit of this system is insufficient for detecting the biomarkers of cancers (9) and diabetes. (10) To realize the highly sensitive detection, improvement of the following points is required.…”

Section: Evaluation Of Antigen Concentration Dependence Using Cartridmentioning

confidence: 99%

Development of Cartridge-Based Wash-Free Single-Step Plasmonic Enzyme-Linked Immunosorbent Assay Using Poly(vinylpyrrolidinone)-Coated Silver Nanoparticles as a Chromogenic Substrate

2017

Sensors and Materials

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In this study, poly(vinylpyrrolidone) (PVP)-coated silver nanoparticles were used as a chromogenic substrate for the establishment of a colorimetric enzyme-linked immunosorbent assay (ELISA) based on localized surface plasmon resonance (LSPR) called "plasmonic ELISA". The PVP-coated silver nanoparticles exhibit a specific color due to the LSPR. In addition, the LSPR absorbance strength of the PVP-coated silver nanoparticles is changed by the aggregation of the silver nanoparticles which depends on the hydrogen peroxide concentration. From these changes in the LSPR absorbance strength in relation to hydrogen peroxide concentration, plasmonic ELISA was established using a glucose oxidase (GOx)-conjugated secondary antibody and glucose for producing hydrogen peroxide by an enzymatic reaction. Furthermore, on the basis of plasmonic ELISA, a cartridge-based single-step plasmonic ELISA device was fabricated for the realization of a wash-free ELISA system. This cartridge-based ELISA system is constructed with a GOxantibody (GOx-conjugated secondary antibody) release pad, a primary antibody immobilized membrane, a glucose release pad, and a PVP-coated silver nanoparticle-containing hydrogel. By using this construction, the antigen concentration can be determined by introducing only a sample solution. As a result, with this cartridge-based ELISA system, the antigen-antibody reaction was successfully detected at different concentrations of antigen (0.1-10 μg/ml).

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Comparison of immunohistochemistry with immunoassay (ELISA) for the detection of components of the plasminogen activation system in human tumour tissue

Cited by 41 publications

References 24 publications

Influence of preoperative core biopsies on uPA/PAI-1 expression in breast cancer tissue

Influence of preoperative core biopsies on uPA/PAI-1 expression in breast cancer tissue

The Myofibroblast Is the Predominant Plasminogen Activator Inhibitor-1-Expressing Cell Type in Human Breast Carcinomas

Development of Cartridge-Based Wash-Free Single-Step Plasmonic Enzyme-Linked Immunosorbent Assay Using Poly(vinylpyrrolidinone)-Coated Silver Nanoparticles as a Chromogenic Substrate

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