A colorimetric immunoassay for the detection of E-cadherin and carcinoembryonic antigen (CEA) expression on human colon carcinoma cell lines in vitro

Eckert, K; Fuhrmann-Selter, Tania; Maurer, H. R.; Büttner, Petra

doi:10.1016/0304-3835(96)04205-x

Cited by 12 publications

(12 citation statements)

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“…So far, cytokine and drug effects on mammary epithelial cell differentiation were mainly analyzed on the basis of morphological changes [29], CK profiles [4,30], lipid accumulation [19], the secretion of milk proteins [20,31] and various differentiation markers [13,32]. In this study, a semiquantitative marker analysis was performed using a colorimetric immunoassay based on the avidin-biotin peroxidase reaction [9] in conjunction with the widely used immunofluorescence technique. With respect to the basal expression of CK19, H type 2 and isoactin on HMEC cells, results were similar to those obtained earlier using immunocytochemistry [2].…”

Section: Discussionmentioning

confidence: 99%

“…The avidin-biotin peroxidase method was performed as described previously [9]. Briefly, cells were grown in 96-well microtiter plates in the presence or absence of the agents for 7 days.…”

Section: Immunocytochemistrymentioning

confidence: 99%

“…Furthermore, the cytokeratin profiles of the lumenal epithelium of benign breast disease and tumor cells are similar in most carcinomas [4]. With respect to the growing interest in differentiation therapy, the use of drugs and cytokines, capable to induce tumor cell differentiation, has been envisaged as a treatment strategy [5][6][7]. However, evaluation of such drugs requires in vitro models that are representative of clinical tumors.…”

Section: Introductionmentioning

confidence: 99%

“…However, evaluation of such drugs requires in vitro models that are representative of clinical tumors. There are few in vitro systems available for the study of human epithelial cell differentiation [8][9][10]. Besides HMEC, immortalized H184 A1N4 cells, derived from cultured nontransformed HMEC, and the tumorigenic CAL51 mammary epithelial cells were only partially characterized with respect to epithelial markers [11,12].…”

Section: Introductionmentioning

confidence: 99%

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Effects of Differentiation Inducers on Cell Phenotypes of Cultured Nontransformed and Immortalized Mammary Epithelial Cells: A Comparative Immunocytochemical Analysis

Grünberg¹,

Eckert²,

Karsten³

et al. 2000

Tumor Biology

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We examined the effects of the differentiation-inducing agents 1,25-dihydroxyvitamin D₃ (calcitriol), phorbol myristate acetate (PMA), the cytokine interferon-γ (IFN-γ), and the tumor necrosis factor-α (TNF-α) alone and in combination with calcitriol on cell growth and differentiation parameters of cultured nontransformed human mammary epithelial cell (HMEC) lines, the chemically transformed HMEC line H184 A1N4, and the human mammary carcinoma cell line CAL51. Cell differentiation was phenotyped by semiquantitative immunocytochemistry using a panel of 15 monoclonal antibodies against marker molecules representing epithelial cell differentiation, cell-cell adhesion processes and malignancy. Cell proliferation of HMEC and H184 A1N4, but not of CAL51 cells was reduced by the agents. Cell phenotypes were analyzed by examining the expression of cytokeratins (pan CK and CK19), the epithelial mucin (MUC1), isoactin, and the blood group-related H type 2 carbohydrate antigen. HMEC and H184 A1N4 cells showed characteristics of basal cells, whereas CAL51 cells resembled a lumenal phenotype. The cell-cell adhesion molecules E-cadherin, intracellular adhesion molecule (ICAM-1), and the tumor markers carcinoembryonic antigen (CEA) and CD44v6 were expressed on all 3 mammary cell lines, with moderate differences. With respect to effects on cell phenotypes, HMEC were sensitive to PMA and IFN-γ resulting in an increased expression of MUC1, CEA and ICAM-1 molecules. H184 A1N4 cells responded to TNF-α in combination with calcitriol by increased expression of pan CK, MUC1, and decreased H type 2 antigen expression, suggesting a transition towards a lumenal phenotype. Furthermore, CEA, ICAM-1 and CD44v6 were increased by TNF-α plus calcitriol. In contrast, CAL51 cells were overall less sensitive to differentiation induction attempts; only TNF-α stimulated MUC1, isoactin and epithelial cell adhesion molecule (Ep-CAM) expression.

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Section: Discussionmentioning

confidence: 99%

“…The avidin-biotin peroxidase method was performed as described previously [9]. Briefly, cells were grown in 96-well microtiter plates in the presence or absence of the agents for 7 days.…”

Section: Immunocytochemistrymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Effects of Differentiation Inducers on Cell Phenotypes of Cultured Nontransformed and Immortalized Mammary Epithelial Cells: A Comparative Immunocytochemical Analysis

Grünberg¹,

Eckert²,

Karsten³

et al. 2000

Tumor Biology

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show abstract

“…Thereafter, enzyme-linked immunosorbent assays (ELISA) [10,11] and chemiluminescence immunoassay (CLIA) [12,13] were widely used in clinical test and many commercially available assay kits were developed. Besides, colorimetric immunoassay [14], time-resolved fluoroimmunoassay [15,16], and liposome immunoassay [17,18] have also been reported for CEA analysis. In recent years, the electrochemical immunosensor has been one of the attractive analytical tools used in CEA detection due to its simplicity and high sensitivity [19,20].…”

Section: Introductionmentioning

confidence: 99%

Micro-plate magnetic chemiluminescence immunoassay and its applications in carcinoembryonic antigen analysis

Zhang

Zhao

et al. 2009

Sci. China Chem.

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A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluorescein isothiocyanate (FITC)-labeled anti-CEA antibodies, CEA antigens, and horseradish peroxidase (HRP)-conjugated anti-CEA antibodies in micro-plate. The immunomagnetic particles coated with anti-FITC antibodies were used as the solid phase for the immunoassay. The separation procedure was carried out by a magnetic plate adaptor and the luminol-hydrogen peroxide (H 2 O 2 )-HRP system was employed for the chemiluminescence detection. The proposed method combined the advantages of the micro-plate reactor and magnetic particle separation technology with the linear range of 5-250 ng mL 1. The detection limit of CEA was 0.61 ng mL 1. The coefficient of the variation was less than 7% and 13% for intra-assay and inter-assay precision, respectively. Compared with the commercial micro-plate chemiluminescent kit, the proposed method showed a good correlation. micro-plate magnetic chemiluminescence immunoassay, carcinoembryonic antigen, tumor marker

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Chemotherapy‐induced immunogenic modulation of tumor cells enhances killing by cytotoxic T lymphocytes and is distinct from immunogenic cell death

Hodge

Garnett

Farsaci

et al. 2013

Intl Journal of Cancer

249

199

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Certain chemotherapeutic regimens trigger cancer cell death while inducing dendritic cell maturation and subsequent immune responses. However, chemotherapy-induced immunogenic cell death (ICD) has thus far been restricted to select agents. In contrast, several chemotherapeutic drugs modulate antitumor immune responses, despite not inducing classic ICD. In addition, in many cases tumor cells do not die after treatment. Here, using docetaxel, one of the most widely used cancer chemotherapeutic agents, as a model, we examined phenotypic and functional consequences of tumor cells that do not die from immunogenic cell death. Docetaxel treatment of tumor cells did not induce ATP or HMGB1 secretion, or cell death. However, calreticulin exposure was observed in all cell lines examined after chemotherapy treatment. Killing by CEA, MUC-1, or PSA-specific CD8+ CTLs was significantly enhanced after docetaxel treatment. This killing was associated with increases in components of antigen-processing machinery, and mediated largely by calreticulin membrane translocation, as determined by functional knockdown of calreticulin, PERK, or calreticulin-blocking peptide. A docetaxel-resistant cell line was selected (MDR-1+, CD133+) by continuous exposure to docetaxel. These cells, while resistant to direct cytostatic effects of docetaxel, were not resistant to the chemomodulatory effects that resulted in enhancement of CTL killing. Here, we provide an operational definition of “immunogenic modulation,” where exposure of tumor cells to nonlethal/sublethal doses of chemotherapy alters tumor phenotype to render the tumor more sensitive to CTL killing. These observations are distinct and complementary to immunogenic cell death and highlight a mechanism whereby chemotherapy can be used in combination with immunotherapy.

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A colorimetric immunoassay for the detection of E-cadherin and carcinoembryonic antigen (CEA) expression on human colon carcinoma cell lines in vitro

Cited by 12 publications

References 4 publications

Effects of Differentiation Inducers on Cell Phenotypes of Cultured Nontransformed and Immortalized Mammary Epithelial Cells: A Comparative Immunocytochemical Analysis

Effects of Differentiation Inducers on Cell Phenotypes of Cultured Nontransformed and Immortalized Mammary Epithelial Cells: A Comparative Immunocytochemical Analysis

Micro-plate magnetic chemiluminescence immunoassay and its applications in carcinoembryonic antigen analysis

Chemotherapy‐induced immunogenic modulation of tumor cells enhances killing by cytotoxic T lymphocytes and is distinct from immunogenic cell death

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