Influence of cytokines, monoclonal antibodies and chemotherapeutic drugs on epithelial cell adhesion molecule (EpCAM) and LewisY antigen expression

Flieger, Dimitri; Hoff, Anja; Sauerbruch, Tilman; Schmidt‐Wolf, Ingo G.H.

doi:10.1046/j.1365-2249.2001.01435.x

Cited by 33 publications

(26 citation statements)

References 46 publications

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“…To investigate the effect of expression level of the target antigen on cell recovery, HT29 colorectal cancer cells were cultured in the presence or absence of IL-4, a cytokine known to suppress Ep-CAM expression (8). Treated and nontreated cells were spiked into separate aliquots of blood samples from healthy donors.…”

Section: Resultsmentioning

confidence: 99%

“…HT29 cells were grown in McCoy's 5a medium containing 1.5 mM L-glutamine and 10% FBS. For modeling the in vivo situation of decreased epithelial cell adhesion molecule (Ep-CAM) expression of OTCs, HT29 cells were cultured for 3 days in the presence of 30 ng͞ml IL-4 (BD Pharmingen) as described by Fleiger et al (8). IL-4 treatment resulted in a 35% decrease of Ep-CAM expression as determined by direct immunofluorescence staining with anti-Ep-CAM fluorochrome-conjugated antibodies (Miltenyi Biotec, Bergisch Gladbach, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…The deposited cells were fixed in ice-cold methanol for 5 min, rinsed in PBS, and blocked with 20% human AB serum (Nabi Diagnostics, Boca Raton, FL) in PBS at 37°C for 20 min. Slides then were incubated at 37°C for 1 h with a monoclonal anti-pan cytokeratin antibody (Sigma), which recognizes human cytokeratins 1,4,5,6,8,10,13,18, and 19 in immunoblotting. Subsequently, slides were washed in PBS and incubated with a mixture of Alexa Fluor 488 and Rphycoerythrin-conjugated goat anti-mouse antibody (Molecular Probes) at 37°C for 30 min.…”

Section: Methodsmentioning

confidence: 99%

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A rare-cell detector for cancer

Krivacic

Ladányi

Curry

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

322

223

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Although a reliable method for detection of cancer cells in blood would be an important tool for diagnosis and monitoring of solid tumors in early stages, current technologies cannot reliably detect the extremely low concentrations of these rare cells. The preferred method of detection, automated digital microscopy (ADM), is too slow to scan the large substrate areas. Here we report an approach that uses fiber-optic array scanning technology (FAST), which applies laser-printing techniques to the rare-cell detection problem. With FAST cytometry, laser-printing optics are used to excite 300,000 cells per sec, and emission is collected in an extremely wide field of view, enabling a 500-fold speed-up over ADM with comparable sensitivity and superior specificity. The combination of FAST enrichment and ADM imaging has the performance required for reliable detection of early-stage cancer in blood.O ccult tumor cells (OTCs) shed from tumors can travel through the blood stream to anatomically distant sites and form metastatic disease, the major cause of cancer-related death in patients with solid tumors. These disseminated cells are present in circulation in extremely low concentrations, estimated to be in the range of one tumor cell in the background of 10 6 -10 7 normal blood cells and are occult to routine imaging and laboratory studies (1). Automated digital microscopy (ADM) using image analysis for recognition of specifically labeled tumor cells has been demonstrated to be the most reliable method currently available for OTC detection (2-5).However, at the typical scan rate of 800 cells per sec, ADM is too slow to screen for a statistically valid number of OTCs (6). This slow scan rate is a result of two factors. One is the substantial latency associated with stepping the sample under the microscopy objective. This stepping results from the lens' small field of view. The other factor is the long exposure time that is due to the low level of excitation from broadband illumination sources and the lack of sensitivity of the charge-coupled device detector used for imaging.Here we report a scanning instrument using fiber-optic array scanning technology (FAST) that can locate OTCs at a rate that is 500 times faster than ADM, with comparable sensitivity and improved specificity. The exposure time is reduced by using a laser source for higher illumination levels and a more sensitive photomultiplier detector. However, our key innovation is providing an optical system with an exceptionally large field of view (50 mm) without a loss of collection efficiency. By collecting the fluorescence in an array of optical fibers that forms a wide collection aperture, the FAST cytometer has a 100-fold increase in field of view over ADM. Although this increase in field of view comes with a reduction in instrument resolution, the resolution is still sufficient for the identification of fluorescently labeled cells. This field of view is large enough to eliminate the need to step the sample under the collection optics, and hence there is no s...

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

A rare-cell detector for cancer

Krivacic

Ladányi

Curry

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

322

223

View full text Add to dashboard Cite

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“…Downregulation of cadherins in turn can upregulate Ep-CAM expression. Moreover, cytokines such as IFNa have been shown to upregulate both COX-2 and Ep-CAM expression in epithelial tumour cells (Bostrom et al, 2001;Flieger et al, 2001). Notably, no correlation was found between Her-2/neu and COX-2 overexpression.…”

Section: Discussionmentioning

confidence: 99%

Correlation of COX-2 and Ep-CAM overexpression in human invasive breast cancer and its impact on survival

et al. 2003

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Recent studies have demonstrated cyclooxygenase 2 (COX-2) overexpression in various human malignancies, especially in breast cancer, where COX-2 turned out to be a predictor of poor survival. To evaluate the relation of COX-2 and Ep-CAM overexpression and its prognostic significance, we performed a retrospective study on 212 breast cancer patients with a median follow-up time of 10.5 years. Overexpression of COX-2 in tumour tissue samples was assessed by immunohistochemistry. COX-2 overexpression was found in 48.6% of the tumour samples and was predictive for poor disease-free and overall survival. Univariate analysis revealed a strong correlation between COX-2 and Ep-CAM overexpression (P ¼ 0.009). Concurrent COX-2 and Ep-CAM overexpression was present in 21.7% of tumour specimens and had an additive negative impact on disease-free and overall survival. Determination of both tumour markers should help in guiding new therapeutic strategies in patients with invasive breast cancer. Cyclooxygenase-2 (COX-2) is a prostaglandin synthase that catalyses the synthesis of prostaglandin G 2 (PGG 2 ) and PGH 2 from arachidonic acid. Recent studies have led to the recognition of the importance of COX-2 in tumorigenesis of different tumour types. It has been shown that COX-2 is involved in tumour angiogenesis (Tsujii et al, 1998;Gately, 2000), in suppression of apoptosis (Sheng et al, 1998) and in the promotion of invasiveness (Tsujii et al, 1997). COX-2 overexpression was found in pancreatic (Molina et al, 1999;Okami et al, 1999;Kokawa et al, 2001), oesophageal (Zimmermann et al, 1999), prostate (Yoshimura et al, 2000), lung (Khuri et al, 2001), head and neck cancers (Chan et al, 1999) and in malignant gliomas (Shono et al, 2001). Tsujii et al reported that COX-2 overexpression in intestinal epithelial cells leads to downregulation of adhesion molecules (i.e. cadherins), resulting in an enhanced tumorigenic potential (Tsujii and DuBois, 1995).Enhanced COX-2 expression in breast cancer was first indicated by reports of elevated prostaglandin levels in breast carcinomas (Bennett et al, 1977), particularly in patients with metastatic disease (Rolland et al, 1980). A key role of COX-2 for the initiation and progression of breast cancer is suggested by the finding that mere overexpression of COX-2 can be sufficient for inducing mammary gland tumorigenesis in transgenic mice (Liu et al, 2001). Notably, in human breast cancer cell lines, a positive correlation was found between invasiveness, metastatic potential and prostaglandin production (Liu and Rose, 1996). Different groups have described the prognostic significance of COX-2 overexpression in breast cancer (Hwang et al, 1998;Ristimaki et al, 2002;Soslow et al, 2000).We have recently described the prognostic significance of Ep-CAM overexpression in patients with invasive breast cancer (Gastl et al, 2000). Ep-CAM (also called 17-1A, ESA, EGP40, 323/A3) is a 40-kDa transmembrane glycoprotein expressed on most human epithelial cells (Gottlinger et al, 1986). The Ep-CAM glycoprote...

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“…Epithelial cell adhesion molecule gene expression appears to be negatively regulated by TNF-alpha through activation of NF-kappaB (Gires et al, 2001). Upon cell cycle arrest by various chemotherapeutics, Ep-CAM surface expression is enhanced (Flieger et al, 2001;Thurmond et al, 2003). As Ep-CAM is involved in adhesion, differentiation and cell proliferation, an influence of Ep-CAM expression on survival of cancer patients can be expected.…”

mentioning

confidence: 99%

Frequent high-level expression of the immunotherapeutic target Ep-CAM in colon, stomach, prostate and lung cancers

et al. 2006

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Epithelial cell adhesion molecule (Ep-CAM; CD326) is used as a target by many immunotherapeutic approaches, but little data are available about Ep-CAM expression in major human malignancies with respect to level, frequency, tumour stage, grade, histologic tumour type and impact on survival. We analysed by immunohistochemical staining tissue microarrays with 4046 primary human carcinoma samples from colon, stomach, prostate and lung cancers for both frequency and intensity of Ep-CAM expression under highly standardised conditions. A total of 3360 samples were analysable. High-level Ep-CAM expression was observed in 97.7% (n ¼ 1186) of colon, 90.7% of gastric (n ¼ 473), and 87.2% of prostate cancers (n ¼ 414), and in 63.9% of lung cancers (n ¼ 1287). No detectable Ep-CAM staining was found with only 0.4% of colon, 2.5% of gastric, 1.9% of prostate cancers, and 13.5% of lung cancers. The only significant correlation of Ep-CAM expression with tumour grading was observed in colon cancer where high-level Ep-CAM expression on grade 3 tumours was down to 92.1% (Po0.0001). Adenosquamous and squamous carcinomas of the lung had a lower percentage of high-level Ep-CAM expression compared to adenocarcinomas with 35.4 and 53.6%, respectively, and with 45.5 and 17.3% of tumours being Ep-CAM negative. With the exception of moderately differentiated colon carcinoma, where patients not expressing Ep-CAM on their tumours showed an inferior survival (P ¼ 0.0014), correlation of Ep-CAM expression with survival did not reach statistical significance for any of the other cancer indications and subgroups. In conclusion, the data strongly support the notion that Ep-CAM is a prime target for immunotherapies in major human malignancies. This is because the most common human cancers show (i) a low frequency of Ep-CAM-negative tumours, (ii) a high frequency of Ep-CAM expression on cells of a given tumour, and (iii) for most cancers, an insignificant influence of tumour staging, grading and histology on Ep-CAM expression.

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Influence of cytokines, monoclonal antibodies and chemotherapeutic drugs on epithelial cell adhesion molecule (EpCAM) and LewisY antigen expression

Cited by 33 publications

References 46 publications

A rare-cell detector for cancer

A rare-cell detector for cancer

Correlation of COX-2 and Ep-CAM overexpression in human invasive breast cancer and its impact on survival

Frequent high-level expression of the immunotherapeutic target Ep-CAM in colon, stomach, prostate and lung cancers

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