CEACAM1 Expression Research Articles

CEACAM6: A HORMONALLY REGULATED PROTEIN IN DIFFERENTIATING HUMAN FETAL LUNG TYPE II CELLS.

Ceacam6 (carcinoembryonic antigen cell adhesion molecule) is a GPI-anchored membrane glycoprotein recently identified in human fetal lung undergoing in vitro hormone-induced differentiation of type II cells. The protein is a tumor biomarker in adult lung and selected other tissues, where it has antimicrobial activity; however, expression and function in lung have not been characterized. Undifferentiated epithelial cells were isolated by collagenase/trypsin digestion from human fetal lung (16-21 wk gestation) and cultured 1-5 d in the absence (control) or presence of DCI (dexamethasone, 10 nM; 8-Br-cAMP, 0.1 mM; isobutylmethylxanthine, 0.1 mM). Some cells in control media were treated with adenovirus Ad12A2 (0.5-6 pfu/cell) to overexpress thyroid transcription factor-1 (TTF-1) or were transfected by electroporation (AMAXA nucleofection technology) with siRNAs (200 nM) for TTF-1 or Ceacam6 to knockdown expression in the presence of DCI. CEACAM6 mRNA and protein were up-regulated ≈10-fold by exposure of fetal lung epithelial cells to either DCI or to a dose of Ad12A2 that produced a level of recombinant TTF-1 comparable with endogenous TTF-1 in DCI-treated cells. By confocal immunofluorescence, CEACAM6 localized to both intracellular vesicles, costaining with SP-B (ie, lamellar bodies), and to the plasma membrane of type II cells. Knockdown of TTF-1 (46 ± 4% of control) in DCI-treated cells reduced CEACAM6 mRNA and protein by 80% (n = 4). Knockdown of CEACAM6 expression by siRNA (88 ± 3%, n = 3) did not alter expression or staining patterns of TTF-1, SP-B or DC-LAMP (a lamellar body membrane protein). Secretion rates for CEACAM6 were similar for both DCI- and Ad12A2-treated cells, with no response to treatment with secretagogues that stimulate surfactant secretion. Treatment of cells with mannosamine, an inhibitor of GPI anchor synthesis, blocked ≈50% of CEACAM6 secretion, consistent with GPI-anchor involvement in the secretory mechanism. We conclude that CEACAM6 expression in primary fetal lung epithelial cells is regulated by glucocorticoid and cAMP and is dependent on TTF-1. CEACAM6 is associated with intracellular surfactant but is secreted primarily by a constitutive pathway independent of lamellar bodies. CEACAM6 may contribute to the innate defense mechanisms of the mature type II cell.

A HUMANIZED PEGYLATED SINGLE CHAIN FRAGMENT VARIABLE (SCFV) TARGETING CARCINOEMBRYONIC ANTIGEN CELL ADHESION MOLECULE 6 REDUCES TUMOR BURDEN IN A SCID MOUSE MODEL BOTH ALONE AND IN COMBINATION WITH GEMCITABINE BY INCREASING APOPTOSIS AND DECREASING PROLIFERATION.

Pancreatic ductal adenocarcinoma is the fourth leading cause of cancer and death in North America. In the year 2002, adenocarcinoma of the exocrine pancreas accounted for ≈30,300 new ases and 29,700 deaths. The current treatment strategy for localized pancreatic cancer is multimodality therapy combining pancreaticoduodenectomy with postoperative adjuvant chemotherapy and external beam radiation therapy, which results in a median overall survival of 12 to 24 months. For locally advanced and metastatic pancreatic cancer, the median survival is 6 to 12 months with currently available palliative chemotherapy. CEACAM6 (carcinoembryonic antigen cell adhesion molecule 6 [CD66c]) is an oncogene expressed on the cell surface as a GPI-linked glycoprotein comprised of 3 Ig-like domains capable of homophilic and heterophilic (CEACAM 5 and 8) interactions. It is highly expressed in > 90% of patients with pancreatic ductal adenocarcinoma. Deregulated expression of CEACAM6 inhibits differentiation and apoptosis of cells deprived of their anchorage to the extracellular matrix. We have previously demonstrated that CEACAM6-expressing pancreatic cancer cell lines treated with a murine monoclonal antibody (13-1) display reduced cellular viability by MTS assay, an indication of cytotoxicity. A murine scFv based on mAb (13-1) as well as a humanized form of the murine scFv also showed in vitro activity in CEACAM6-expressing cancer cell lines. In vivo studies using an SCID mouse model xenograft with BXPC-3 cells indicated a significant decrease in tumor burden as a result of treatment with a PEGylated form of the previously mentioned humanized scFv. The humanized scFv drug reduced the tumor burden as a stand-alone agent and in combination with gemcitabine compared with the control animal groups. Immunohistochemical analysis of tumors excised from animals treated with the humanized scFv and untreated animals indicated a marked increase in apoptosis (H&E) as well as a marked decrease in proliferation (Ki67) in the treated group relative to the untreated group. This observed in vivo effect on proliferation and apoptosis is currently being confirmed in vitro using CEACAM6-expressing cancer cell lines.

Exercise training increases hepatic CEACAM1 expression

The low expression level of hepatic carcinoembryonic antigen-related cell adhesion molecule 1 (hCEACAM1), responsible for circulating-insulin clearance, is associated with insulin resistance. The purpose of this study was to determine if hCEACAM1 expression would be increased and associated with altered glucose tolerance after training, and its training-induced changes would depend on genetic factors. Two different inbred rat strains [DA & Copenhagen (COP)] were divided into control and training groups. Rats in the training group underwent a swim training regimen consisting of two 3-hour long bouts separated by a 45-min long rest period, 6 days/wk for 4 weeks. At the completion of the training, all rats underwent a glucose tolerance test where blood glucose levels were measured at 6 time points over a 75-min interval following i.p. glucose injection (2g/kg BW). hCEACAM1 expression levels analyzed by Western blotting increased by 85% (P<0.05) in DA, but not in COP, rats after training. Improvement in training-induced glucose tolerance was also greater in DA (58%, P<0.001) than in COP (25%, P<0.01). The results demonstrate that hCEACAM1 is upregulated with endurance training and may play a role in training-induced improvement in glucose tolerance and the magnitude of training-induced change in hCEACAM1 expression level is dependent on genetic factors. Supported by Ohio Challenge Fund

CEACAM1 expression in peripheral blood and placental tissue in preeclamptic patients

CEACAM1 expression in peripheral blood and placental tissue in preeclamptic patients

Glycoconjugate profiling of primary melanoma and its sentinel node and distant metastases: Implications for diagnosis and pathophysiology of metastases

Aiming at more precise detection of melanoma cells in sentinel lymph nodes and better understanding of the mechanisms underlying metastatic spread, expression of L1, CEACAM1, and binding of the lectins HPA, ML-I and PNA, was assessed in benign nevi ( n = 12), primary melanomas (PTs: n = 67), their corresponding sentinel lymph nodes (SLNs: n = 40), and distant metastases (DMs: n = 35). Sensitivity and specificity of CEACAM1 (95–97%; 66%) and L1 (90–93%; 100%) exceeded that of the standard markers MelanA, S100, and HMB45 in single marker use. Lectin binding was found in PTs and DMs (HPA: 69% and 77%; ML-I: 82% and 77%, respectively), but rarely in SLNMs (HPA: 20%, ML-I: 20%, PNA: 5%, respectively). The highly specific and sensitive L1-11A against L1 and 4D1/C2 against CEACAM1 antibodies are a worthy completion to standard antibody panels for diagnosis of melanoma cells. Both CAMs seem to be functionally involved in lymphatic and haematogenous spread, and are thus promising target molecules for immunotoxins.

Expression of CEACAM1 in pulmonary adenocarcinoma cells and their metastases

17071 Background: The cell adhesion molecule CEACAM1 is involved in intercellular adhesion and is expressed in a variety of normal human tissues such as mammary gland, colonic mucosa and prostate. In cases of malignant transformation of these tissues a down regulation or loss of CEACAM1 has been shown. In contrary, CEACAM1 is not expressed in normal lung tissue or melanocytes and it has been demonstrated that a expression in these tissues is associated with the development of metastatic disease. The aim of the present investigation was to analyze a possible association between the expression of CEACAM1 in pulmonary adenocarcinoma cells and their lymph node and hematogenous metastatic cells. Methods: CEACAM1 expression was immunhistochemically evaluated in primary tumor cells, cells in lymph nodes and in distant metastases of 96 patients with metastatic adenocarcinoma of the lung who had undergone surgery between 1999 and 2002. Results: An expression of CEACAM1 has been shown in 78 of 96 primary tumors. We found a significant positive correlation between the CEACAM1 expression on the cells of the primary tumor and the lymph node metastases (p&lt;0.005) and the hematogenous metastases (p&lt;0.05). Conclusion: As shown before, CEACAM1 is not expressed in normal lung tissue but in most primary adenocarcinomas of the lung. We are the first to demonstrate, that its expression is preserved in the lymph node and hematogenous metastases in metastatic disease implicating that its expression is of functional significance of both metastatic sites. No significant financial relationships to disclose.

Increased colon tumor susceptibility in azoxymethane treated CEABAC transgenic mice

Human carcinoembryonic antigen (CEA), a widely used clinical tumor marker, and its close relative, CEACAM6, are often overexpressed in many cancers. This correlation suggests a possible instrumental role in tumorigenesis, which is supported by extensive results obtained with several in vitro systems. The implication that these results could also apply in vivo warrants investigation. Since mice do not possess homologs of the glycophosphatidyl inositol (GPI)-anchored CEACAM family genes CEA, CEACAM6 and CEACAM7, we have constructed transgenic mice harboring a 187 kb portion of the human CEACAM family gene locus contained in a bacterial artificial chromosome (CEABAC) that includes genes coding for CEA, CEACAM6 and CEACAM7. In this study, we treated the CEABAC mice and their wild-type littermates with azoxymethane (AOM) in order to induce colon tumor formation. At 20 weeks post-treatment, the CEABAC transgenics showed more than a 2-fold increase in mean tumor load relative to their wild-type littermates. Cell surface expression of CEA and CEACAM6 increased by 2- and 20-fold, respectively, in colonocytes from the tumors relative to colonocytes from non-AOM treated transgenics and a de-regulated spatial pattern of CEA/CEACAM6 expression was found in 'normal' crypts adjacent to the tumors, thus mimicking closely the situation in human colon tumorigenesis. A modestly increased incidence of beta-catenin mutations also observed in the AOM-induced CEABAC tumors. These results show that expression of the human GPI-anchored CEACAM family genes predisposes mice to acquire and/or retain essential mutations necessary for sporadic colon tumor development.

Expression Pattern of Osteopontin in Endometrial Carcinoma: Correlation With Expression of the Adhesion Molecule CEACAM1

Osteopontin (OPN) and CEACAM1 have diverse biological functions in the uterus and placenta throughout the estrous cycle and pregnancy and have been shown to interact with integrin beta3. OPN is a glycoprotein of the extracellular matrix, which has been shown to mediate cellular migration and invasion and to contribute to tumorigenesis in several types of cancers. Recently we showed the expression pattern of OPN in gestational trophoblastic tumors. CEACAM1 is an adhesion molecule of the carcinoembryonic antigen family that we have recently found to be expressed in endometrial cancer and that has been shown to be down-regulated in colorectal, prostate, and breast cancer. In this study, immunohistochemistry and immunofluorescence with specific antibodies were performed on a series of 20 normal endometrial samples, 17 endometrial hyperplasias, and 43 endometrial carcinomas (28 endometrioid, 10 serous, and 5 clear cell carcinomas) to investigate the expression pattern and cell-type specific localization of OPN and to correlate it with the expression of CEACAM1. In addition, Western blot was performed on normal human endometrium and endometrial neoplasia. Strong OPN expression with a consistent cytoplasmic localization in epithelial glandular cells was observed in the normal human endometrium in 80% of the samples of the proliferative and secretory phase (score 8-12). Similar results could be found in endometrial hyperplasias. Strong expression of OPN could be observed in 29 (67.4%) of the 43 analyzed endometrial carcinomas. Of the 43 analyzed tumors, 18 (41.8%) were in the high score (8-12) category with a strong OPN expression level; 11 of 43 (25.5%) showed a moderate score (4-7) category. In endometrioid carcinoma with increasing malignancy grade, increasing areas with low OPN expression level or complete loss of OPN expression could be observed. In contrast, serous tumors showed a strong OPN expression level. Similar results could be found in Western blot analysis. CEACAM1 showed similar results and could be found to be coexpressed with OPN in normal human endometrium and in endometrial neoplasia as we showed using immunofluorescence. In this study, the different expression patterns of OPN in endometrial tumors could additionally support the biological diversity of endometrioid and serous carcinomas together with other markers. We suggest that OPN might play a different role in the pathogenesis of endometrial cancer (possibly as a functional complex with CEACAM1) and could be relevant for invasive growth of such lesions.

Stimulation of CEACAM1 expression by 12-O-tetradecanoylphorbol-13-acetate (TPA) and Calcium Ionophore A23187 in endometrial carcinoma cells and placental hybridoma cells

Stimulation of CEACAM1 expression by 12-O-tetradecanoylphorbol-13-acetate (TPA) and Calcium Ionophore A23187 in endometrial carcinoma cells and placental hybridoma cells

Stimulation of CEACAM1 expression by 12- O -tetradecanoylphorbol-13-acetate (TPA) and calcium ionophore A23187 in endometrial carcinoma cells

Downregulation of carcinoembryonic antigen-related cell adhesion molecule (CEACAM1), a cell adhesion molecule with tumor suppressing properties has been observed in a high percentage of carcinomas of the endometrium and other malignancies. The mechanisms for the dysregulation and the role of hormones and cytokines on the expression of CEACAM1 in endometrial carcinomas is unknown. We therefore studied the effect of estradiol, medroxyprogesterone acetate (MPA), RU486, gamma-interferon (IFN-gamma), tumor necrosis factor alpha (TNF-alpha), 12-O-tetradecanoylphorbol-13-acetate (TPA) and calcium ionophore A23187 on the expression in the non-expressing endometrial tumor cell lines Hec1B and Skut1B, respectively. No induction of CEACAM1 expression was observed in Hec1B endometrial adenocarcinoma cells in response to hormones and cytokines whereas treatment with TPA and calcium ionophore A23187 resulted in the strong expression of endogenous CEACAM1 on the mRNA and protein levels. In contrast, no induction of CEACAM1 expression was observed in endometrial mixed mesenchymal Skut1B cells. Studies of other members of the CEACAM family revealed that the re-expression in Hec1B carcinoma cells is restricted to CEACAM1 suggesting a cell type-specific and cell type-independent mechanism of CEACAM1 activation via the protein kinase C (PKC) pathway. Induction of CEACAM1 expression was dependent on protein kinase C protein synthesis and luciferase reporter assays with CEACAM1 promoter constructs demonstrated that the re-expression of CEACAM1 is regulated at the transcriptional level. This is the first report demonstrating that activators of PKC are able to specifically induce the expression of CEACAM1 in human carcinoma cells and our findings may provide a basis for the therapeutic inhibition of tumor growth in malignancies in which CEACAM1 is downregulated.

Characterization of a Side Population of Cancer Cells from Human Gastrointestinal System

A subset of stem cells, termed "side population" (SP) cells, has been identified and characterized in several mammalian tissues and cell lines. However, SP cells have never been identified or isolated from gastrointestinal cancers. We used flow cytometry and the DNA-binding dye Hoechst 33342 to isolate SP cells from various human gastrointestinal system cancer cell lines. Fifteen of sixteen cancer cell lines from the gastrointestinal system contained 0.3%-2.2% SP cells. Next, we used an oligonucleotide microarray to analyze differentially expressed genes between SP and non-SP cells of hepatoma HuH7. The expression of GATA6, which is associated with embryonic development and hepatocytic differentiation, was significantly upregulated in HuH7 SP cells. The expression of ABCG2, ABCB1, and CEACAM6, which are associated with chemoresistance, was also significantly increased in SP cells. In addition, some epithelial markers and mesenchymal markers were overexpressed in SP cells. Reverse transcription-polymerase chain reaction and immunocytochemical staining validated these results and suggested a multilineage potential for HuH7 SP cells. In hepatoma HuH7 and colorectal SW480 cell lines, SP cells showed evidence for self-renewal, generating both SP and non-SP cells. Finally, chemoresistance to anticancer agents, including doxorubicin, 5-fluorouracil, and gemcitabine, were compared between HuH7 SP and non-SP cells using an ATP bioluminescence assay. The HuH7 SP cells expressed a higher resistance to doxorubicin, 5-fluorouracil, and gemcitabine compared with non-SP cells. These findings demonstrate that cancers of the gastrointestinal system do contain SP cells that show some characteristics of so-called stem cells.

Osteopontin Expression in Gestational Trophoblastic Diseases: Correlation With Expression of the Adhesion Molecule, CEACAM1

The human placenta is a complex tissue with multiple endocrine and nutritional functions and a unique capacity for rapid proliferation but tightly controlled invasion, differentiating it from malignant tumors. Osteopontin (OPN) is a glycoprotein of the extracellular matrix, which has been shown to mediate cellular migration and invasion and to contribute to tumorigenesis in several types of cancers. OPN also could be implicated in regulating implantation and placentation by promoting cellular migration and invasion in a placenta-specific fashion. We could demonstrate the expression pattern of OPN in the normal human placenta in which it is localized in the extravillous (intermediate) trophoblast and the villous cytotrophoblast. CEACAM1 is an adhesion molecule, which we have recently found to be expressed at the maternal-fetal interface of the normal placenta with a localization to the extravillous (invasive) trophoblast and in gestational trophoblastic disease (GTD) and also to be potentially implicated in trophoblast invasion and tumorigenesis. Both OPN and CEACAM1 have been shown to interact with integrin beta3. The purpose of this study was to investigate the expression pattern of OPN in GTD and to correlate it with the expression of CEACAM1. To analyze the expression of OPN, we performed immunohistochemistry on a total of 27 cases of GTD, including 21 hydatidiform moles and 6 choriocarcinomas, which had previously been characterized with respect to their CEACAM1 expression. Hydatidiform moles showed a positivity for OPN in villous cytotrophoblast and in the trophoblast proliferations on the villous surface. The strongest OPN expression could be observed in the choriocarcinomas with a heterogenous OPN expression pattern. CEACAM1 had shown similar results and was found to be expressed in choriocarcinoma. The expression pattern of osteopontin in gestational trophoblastic diseases indicates that it might play a role in the pathogenesis of GTD (possibly as a functional complex with CEACAM1 and integrin beta3) and might be useful as an additional diagnostic marker for such lesions.

Osteopontin Is Colocalized with the Adhesion Molecule CEACAM1 in the Extravillous Trophoblast of the Human Placenta and Enhances Invasion of CEACAM1-Expressing Placental Cells

The human placenta is a complex tissue and possesses, through its capacity to proliferate and to invade maternal tissue, qualities that are usually found in malignant tumors. Osteopontin (OPN) and CEACAM1 may regulate these processes. The present study was designed to investigate the expression pattern of OPN in the human placental components and to correlate it with CEACAM1 expression and function in placental cell invasiveness. Immunohistochemistry with an OPN-specific antibody and immunofluorescence were performed on normal placental samples to investigate the expression pattern of OPN and CEACAM1 in the human placenta. Extravillous trophoblast (EVT) hybridoma cells transfected with CEACAM1 and stimulated with OPN were studied using the Matrigel invasion assay. All placentae presented very strong expression of OPN in the EVT at the invasion front, where it colocalized with CEACAM1. In addition, OPN was also present in the villous trophoblast, with strongest expression in the cytotrophoblast of the first trimester. Transfection with CEACAM1 followed by stimulation with OPN resulted in increased invasiveness of EVT hybridoma cells. The present study shows the first systematic analysis of OPN expression pattern in the human placenta showing strong expression in the EVT at the invasion front. Colocalization of OPN with CEACAM1 in the EVT indicates that they might act together to regulate invasiveness at the maternal-fetal interface. Using an in vitro model, we also demonstrated increased cellular invasiveness after OPN treatment. We speculate that OPN and CEACAM1 may act as a functional complex involved in the regulation of placental invasiveness.

The involvement of NK cells in ankylosing spondylitis

A role for NK cells in the regulation of autoimmunity has been demonstrated. Since there is a strong association between Ankylosing Spondylitis (AS) and HLA-B27, which is specifically recognized by the NK-inhibitory receptor KIR3DL1, this study evaluated the potential involvement of NK cells in AS. We studied 19 AS patients and 22 healthy volunteer donors and assessed the percentage, activity and receptor expression of peripheral blood NK cells. We also evaluated candidate-inflammatory mediators in sera. We found that AS patients have significantly higher percentages of NK cells. However, we found no differences between the ability of NK cells derived from AS and healthy controls to recognize target cells expressing HLA-B27. Remarkably, we observed that the NK-inhibitory receptor CEACAM1 (carcino-embryonic antigen-cell adhesion molecule) is highly expressed among AS-derived NK cells. Furthermore, engagement of CEACAM1 inhibited NK activity in these patients. Finally, we demonstrated that CEACAM1 expression is induced by IL-8 and SDF-1 (stromal cell derived factor), both of which are present in high levels in the sera of AS patients. These results may indicate that NK cells and CEACAM1 play a role in AS pathogenesis and implicate chemokines in the mechanism of CEACAM1 expression.

CEACAM1 (CD66a) mediates delay of spontaneous and Fas ligand-induced apoptosis in granulocytes

Granulocytes form the first and fastest line of defense against pathogenic infections. Their survival is limited by apoptosis, a process that is critical for the resolution of inflammation. Pro-apoptotic and pro-inflammatory cytokines, as well as several receptors, can alter the lifespan of granulocytes. Here we report that the carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM1, CD66a) is involved in the regulation of granulocyte survival. Until now CEACAM1 is described to control cell proliferation, cell migration, tumor growth, angiogenesis and diverse leukocyte functions. However, very little is known about its role in granulocytes. We found that CEACAM1 expression in resting rat granulocytes is significantly higher than in other leukocyte subtypes. Stimulation led to a strongly increased CEACAM1 cell surface expression and to release of soluble CEACAM1. DNA fragmentation assays and annexin V staining revealed that binding of CEACAM1-specific antibodies, Fab fragments and soluble CEACAM1-Fc constructs to cell surface-expressed CEACAM1 causes a delay of spontaneous and Fas ligand (CD95L)-induced apoptosis. Tyrosine phosphorylation of CEACAM1-L, its association with SHP-1, the activation of Erk1/2 and caspase-3 appeared to be crucial for the CEACAM1-mediated anti-apoptotic effect. These findings provide evidence that CEACAM1 influences the resolution of inflammation by prolonging the survival of rat granulocytes.

CEACAM1 expression in intraductal papillary mucinous neoplasms (IPMNs) of the pancreas: A factor of prognostic significance in invasive IPM-carcinomas

CEACAM1 expression in intraductal papillary mucinous neoplasms (IPMNs) of the pancreas: A factor of prognostic significance in invasive IPM-carcinomas

Expression pattern of osteopontin (OPN) in the human placenta – correlation with expression of the adhesion molecule CEACAM1

The human placenta is a complex tissue with multiple endocrine and nutritional functions and a unique capacity for rapid proliferation but tightly controlled invasion, differentiating it from malignant tumors. The cytokine osteopontin (OPN) is a glycoprotein of the extracellular matrix which has been shown to mediate cellular migration and invasion and to contribute to tumorigenesis in several types of cancers. OPN could thus also be implicated in regulating implantation and placentation by promoting cellular migration and invasion in a placenta-specific fashion. The expression pattern and cell-type specific localization of OPN in the human placenta has not been described so far. CEACAM1 is an adhesion molecule which we have recently found to be expressed at the maternal-fetal interface with a localization to the extravillous (invasive) trophoblast and to be potentially implicated in trophoblast invasion. Both OPN and CEACAM1 have been shown to interact with integrin beta3. In the present study, immunohistochemistry with specific antibodies was performed to investigate the expression pattern and cell-type specific localization of OPN in the human placenta, and to correlate it with the expression of CEACAM1. Parallel sections of 50 normal placentas where analysed for OPN and CEACAM1 expression. All placentas presented very strong expression of OPN in the extravillous (invasive) trophoblast (80% positive cells), which correlated with expression of CEACAM1. In addition, OPN was also present in the villous trophoblast, with strongest expression in the cytotrophoblast of the first trimester, while CEACAM1 was not expressed in the villous trophoblast. These data show that OPN is specifically expressed in the human placenta with strongest expression levels in the extravillous (invasive) trophoblast, where it correlates with expression of CEACAM1 and could thus contribute in the regulation of trophoblast invasion in a CEACAM1-involving mechanism as well as play an additional role in cytotrophoblast function.

Immunobead multiplex RT-PCR detection of carcinoembryonic genes expressing cells in the blood of colorectal cancer patients

Circulating cell detection using reverse transcriptase-polymerase chain reaction (RT-PCR) techniques has been studied as a new prognostic factor in colorectal cancer patients. With the view of enhancing detection sensitivity, we developed a new multiplex RT-PCR assay for circulating cell detection based on the expression of carcinoembryonic antigen-related cell adhesion molecule 5 (CEACAM5; formerly CEA) and CEACAM7 (formerly CGM2). Between November 2002 and December 2003, 45 stage III-IV, 39 stage I-II colorectal cancer patients, 32 non-colorectal cancer patients and 41 healthy individuals were included. Positive selection using HEA-125 immunobeads was applied to blood samples before mRNA extraction, cDNA synthesis and a multiplex CEACAM5/CEACAM7 RT-PCR assay. For both CEACAM5 and CEACAM7, the limit of detection was found to be as low as 1 expressing cell in 10(6) nucleated blood cells. The multiplex RT-PCR assay was negative for the 41 healthy individuals and the 32 non-colorectal cancer patients. The test was positive in 53/84 (63%) of the colorectal cancer patients for CEACAM5 and/or CEACAM7, whereas 32/84 (38%) were positive for both markers. Colorectal cancer patients were positive for one of the two markers in 80% of cases (36/45) for stage III-IV patients (CEACAM5 73%, CEACAM7 51%) and in 44% of cases (17/39) for stage I-II patients. This multiplex RT-PCR assay with two markers proved to be more sensitive than use of a single marker in detecting circulating tumour cells. The discrepant expression of CEACAM5 and CEACAM7 may label circulating tumour cells that have different levels of differentiation and subsequent aggressive behaviour.

The human tumor suppressor CEACAM1 modulates apoptosis and is implicated in early colorectal tumorigenesis

Defects in the adenomatous polyposis coli (APC) tumor suppressor pathway are sufficient for neoplastic transformation as the initiating step in colorectal carcinogenesis. In contrast, hyperplastic tumors possess normal APC function, and it is unclear whether they represent significant precursor lesion in cancer development. CEACAM1 is a tumor suppressor whose expression is known to be lost in the great majority of early adenomas and carcinomas. We found that loss of CEACAM1 expression is more common in neoplastic tumors than APC mutations. While APC function was normal in hyperplastic aberrant cypt foci and hyperplastic polyps, loss of CEACAM1 was observed as frequently as in the neoplasias. Moreover, the presence or absence of CEACAM1 expression in the hyperplastic tumors correlates with normal or reduced apoptosis, respectively. In vitro, CEACAM1 acts as a regulator of apoptosis in CEACAM1-transfected Jurkat cells. Finally, in human HT29 colon cancer cells, apoptosis can be specifically restored by induction of CEACAM1 expression. These data suggest an oncodevelopmental link between neoplasia and hyperplasia and demonstrate that CEACAM1 acts as a regulator of apoptosis in the colonic epithelium. Thus, failure of the maturing colon cell to express CEACAM1 is likely to contribute to the development of hyperplastic lesions, which may eventually pave the way to neoplastic transformation and colon cancer development.

CEACAM1 Enhances Invasion and Migration of Melanocytic and Melanoma Cells

Expression of the cell adhesion molecule CEACAM1 in melanomas is an independent factor for the risk of metastasis with a predictive value superior to that of tumor thickness. We have previously shown that CEACAM1 co-localizes at the tumor-stroma interface of invading melanoma masses with integrin beta(3) and that these two adhesion molecules interact via the CEACAM1 cytoplasmic domain. To address the functional consequences of CEACAM1 expression, we investigated invasion and migration of melanocytic and melanoma cells that stably express CEACAM1 using two different in vitro systems. Here, we demonstrate that CEACAM1 expression markedly enhances cell invasion and migration. The enhanced invasion and migration of CEACAM1-transfected cells was dependent on the presence of Tyr-488 within the full-length cytoplasmic CEACAM1 domain. Treatment with anti-CEACAM monoclonal antibodies blocked CEACAM1-enhanced cell invasion and cell migration in a dose-dependent manner. Furthermore, the enhanced invasion and migration of CEACAM1-transfected melanoma cells was blocked by integrin-antagonizing RGD peptides. Expression of integrin beta(3) induces the up-regulation of CEACAM1 in melanocytic MEL6 cells. These results strengthen the view that CEACAM1 and alpha(v)beta(3) integrin are functionally interconnected with respect to the invasive growth of melanomas.