alpha-Neup5Ac-(2--3)-beta-D-Galp-(1--4)-[alpha-L-Fucp-(1--3)]-D-GlcpNAc and Lung-Neoplasms

Serum biomarkers provide valuable information about the diagnosis and prognosis of a wide variety of malignant tumors. Despite the identification of several useful serum biomarkers in lung cancer, consensus on their utility has not yet been reached. Furthermore, guidelines and standard protocols to implement their use for patients with lung cancer are lacking, despite the accumulation of much data on the efficacy of several serum biomarkers over recent decades. In this review, we discuss the molecular features, functions, and clinical relevance of the conventional serum biomarkers for lung cancer, including carcinoembryonic antigen (CEA), cytokeratin 19 fragment 21-1 (CYFRA 21-1), tissue polypeptide antigen (TPA), carbohydrate antigen 19-9 (CA19-9), sialyl Lewis

Topics: Adenocarcinoma; Biomarkers, Tumor; CA-125 Antigen; CA-19-9 Antigen; Carcinoembryonic Antigen; Carcinoid Tumor; Carcinoma, Neuroendocrine; Carcinoma, Squamous Cell; Humans; Keratin-19; Lung Neoplasms; Neuroendocrine Tumors; Oligosaccharides; Peptide Fragments; Phosphopyruvate Hydratase; Recombinant Proteins; Sialyl Lewis X Antigen; Tissue Polypeptide Antigen

Tumor markers are defined as substances which are produced by cancer cells or non-cancer cells reactive to cancer cells, and reflect the cancer status, such as its presence, characteristics, and volume. Clinically, many tumor markers are useful not only to assess the presence/absence of cancer, the primary site, histology, stage, and recurrence, but also to monitor the anti-cancer therapy. Tumor markers for lung cancer play only supporting roles because of their limited sensitivity and specificity, but they are clinically essential to daily medical oncology. This review addresses 6 important tumor markers for lung cancer, namely, CEA, SLX, CYFRA, SCC, ProGRP, and NSE.

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Humans; Keratins; Lung Neoplasms; Oligosaccharides; Peptide Fragments; Peptides; Phosphopyruvate Hydratase; Recombinant Proteins; Serpins; Sialyl Lewis X Antigen

We investigated the immunohistochemical expression of three carbohydrate antigens, sialyl-Lewisa, sialyl-Lewisx and sialyl-Lewisx-i, in human lung cancer tissues using monoclonal antibodies, 2D3, SNH3 and FH6, respectively, and compared the expression rate of these three antigens with clinical and pathologic findings. The expression rate of all the three antigens in adenocarcinoma was higher than that in squamous cell carcinoma and that of sialyl-Lewisx was highest in adenocarcinoma. Sialyl-Lewisx antigen was expressed in all cases of positive nodal metastasis or postoperative distant metastasis in adenocarcinoma. In squamous cell carcinoma, however, there was no relationship between the expression rate of sialyl-Lewisx antigen and nodal or distant metastasis. These results suggest that expression of sialyl-Lewisx antigen in adenocarcinoma of the lung may be correlated with nodal or distant metastasis.

Topics: Adenocarcinoma; Antigens, Tumor-Associated, Carbohydrate; CA-19-9 Antigen; Carcinoma, Squamous Cell; Gangliosides; Humans; Lung Neoplasms; Lymphatic Metastasis; Oligosaccharides; Sialyl Lewis X Antigen

Cancer-related carbohydrate epitopes, which are regarded as potential diagnostic and prognostic biomarkers, are carried on the main acute phase proteins. It is not clear, however, if the glycosylation profile is similar in different glycoproteins, or it is protein specific to some extent. The aim of the study was to compare fucosylation, α2,3 sialylation and expression of sialyl-Lewisx epitopes (sLe(x)) in the serum as a whole, AGP and haptoglobin of small cell (SCLC) and non-small cell lung cancer (NSCLC) patients with respect to healthy subjects as well as the cancer stage and its histological type.. Thirty-three NSCLC, 13 SCLC patients and 20 healthy volunteers were included in the study. Carbohydrate epitopes were detected by means of their reactivity with specific lectins and monoclonal anti-sLe(x) antibodies in direct or dual-ligand ELISA tests.. Significantly increased fucosylation was found in total serum in both cancer groups and in NSCLC haptoglobin. No difference was observed in SCLC haptoglobin or α₁-acid glycoprotein in both cancer groups. Also α2,3 sialylation was elevated in total serum, but not in α₁-acid glycoprotein. This type of sialylation was undetectable in haptoglobin by means of MAA reactivity, in both healthy and cancer subjects. Complete sLe(x) antigens were overexpressed in total NSCLC serum and SCLC AGP, and their level was considerably lowered in cancer haptoglobin.. Typical acute phase proteins, haptoglobin and AGP, exhibit different glycosylation profiles in lung cancer. Alterations observed in haptoglobin reflected the disease process better than those in AGP. Comparison of haptoglobin and AGP glycosylation to that observed in total serum suggests that some efficient carriers of disease-altered glycoproteins still remain unidentified.

Topics: Biomarkers; Carcinoma, Non-Small-Cell Lung; Glycosylation; Haptoglobins; Healthy Volunteers; Humans; Lung Neoplasms; Neoplasm Staging; Oligosaccharides; Orosomucoid; Prognosis; Sialyl Lewis X Antigen; Small Cell Lung Carcinoma

Extravasation of circulating tumor cells (CTCs) is critical for metastasis and is initiated by adhesive interactions between glycoligands on CTCs and E-selectin on endothelia. Here, we show that the clinically approved proteasome inhibitor bortezomib (BZM; Velcade) counteracts the cytokine-dependent induction of E-selectin in the lung mediated by the primary tumor, thereby impairing endothelial adhesion and thus spontaneous lung metastasis in vivo. However, the efficacy of BZM crucially depends on the tumor cells' E-selectin ligands, which determine distinct adhesion patterns. The canonical ligands sialyl-Lewis A (sLeA) and sLeX mediate particularly high-affinity E-selectin binding so that the incomplete E-selectin-reducing effect of BZM is not sufficient to disrupt adhesion or metastasis. In contrast, tumor cells lacking sLeA/X nevertheless bind E-selectin, but with low affinity, so that adhesion and lung metastasis are significantly diminished. Such low-affinity E-selectin ligands apparently consist of sialylated MGAT5 products on CD44. BZM no longer has anti-metastatic activity after CD44 knockdown in sLeA/X-negative tumor cells or E-selectin knockout in mice. sLeA/X can be determined by immunohistochemistry in cancer samples, which might aid patient stratification. These data suggest that BZM might act as a drug for inhibiting extravasation and thus distant metastasis formation in malignancies expressing low-affinity E-selectin ligands.

Topics: Animals; Bortezomib; CA-19-9 Antigen; Cell Adhesion; E-Selectin; Humans; Ligands; Lung Neoplasms; Mice; Neoplasm Metastasis; Oligosaccharides; Sialyl Lewis X Antigen

The formation of distant metastasis resulting from vascular dissemination is one of the leading causes of mortality in non‑small cell lung cancer (NSCLC). This metastatic dissemination initiates with the adhesion of circulating cancer cells to the endothelium. The minimal requirement for the binding of leukocytes to endothelial E‑selectins and subsequent transmigration is the epitope of the fucosylated glycan, sialyl Lewis x (sLex), attached to specific cell surface glycoproteins. sLex and its isomer sialyl Lewis a (sLea) have been described in NSCLC, but their functional role in cancer cell adhesion to endothelium is still poorly understood. In this study, it was hypothesised that, similarly to leukocytes, sLe glycans play a role in NSCLC cell adhesion to E‑selectins. To assess this, paired tumour and normal lung tissue samples from 18 NSCLC patients were analyzed. Immunoblotting and immunohistochemistry assays demonstrated that tumour tissues exhibited significantly stronger reactivity with anti‑sLex/sLea antibody and E‑selectin chimera than normal tissues (2.2‑ and 1.8‑fold higher, respectively), as well as a higher immunoreactive score. High sLex/sLea expression was associated with bone metastasis. The overall α1,3‑fucosyltransferase (FUT) activity was increased in tumour tissues, along with the mRNA levels of FUT3, FUT6 and FUT7, whereas FUT4 mRNA expression was decreased. The expression of E‑selectin ligands exhibited a weak but significant correlation with the FUT3/FUT4 and FUT7/FUT4 ratios. Additionally, carcinoembryonic antigen (CEA) was identified in only 8 of the 18 tumour tissues; CEA‑positive tissues exhibited significantly increased sLex/sLea expression. Tumour tissue areas expressing CEA also expressed sLex/sLea and showed reactivity to E‑selectin. Blot rolling assays further demonstrated that CEA immunoprecipitates exhibited sustained adhesive interactions with E‑selectin‑expressing cells, suggesting CEA acts as a functional protein scaffold for E‑selectin ligands in NSCLC. In conclusion, this work provides the first demonstration that sLex/sLea are increased in primary NSCLC due to increased α1,3‑FUT activity. sLex/sLea is carried by CEA and confers the ability for NSCLC cells to bind E‑selectins, and is potentially associated with bone metastasis. This study contributes to identifying potential future diagnostic/prognostic biomarkers and therapeutic targets for lung cancer.

Topics: Aged; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Cell Adhesion; E-Selectin; Female; Fucosyltransferases; Humans; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Polysaccharides; Sialyl Lewis X Antigen

We have recently elucidated a novel function for CD82 in E-cadherin-mediated homocellular adhesion; due to this function, it can inhibit cancer cell dissociation from the primary cancer nest and limit metastasis. However, the effect of CD82 on selectin ligand-mediated heterocellular adhesion has not yet been elucidated. In this study, we focused on the effects of the metastasis suppressor CD82/KAI1 on heterocellular adhesion of cancer cells to the endothelium of blood vessels in order to further elucidate the function of tetraspanins. The over-expression of CD82 in cancer cells led to the inhibition of experimentally induced lung metastases in mice and significantly inhibited the adhesion of these cells to human umbilical vein epithelial cells (HUVECs) in vitro. Pre-treatment of the cells with function-perturbing antibodies against sLea/x significantly inhibited the adhesion of CD82-negative cells to HUVECs. In addition, cells over-expressing CD82 exhibited reduced expression of sLea/x compared to CD82-negative wild-type cells. Significant down-regulation of ST3 β-galactoside α-2, 3-sialyltransferase 4 (ST3GAL4) was detected by cDNA microarray, real-time PCR, and western blotting analyses. Knockdown of ST3GAL4 on CD82-negative wild-type cells inhibited expression of sLex and reduced cell adhesion to HUVECs. We concluded that CD82 decreases sLea/x expression via the down-regulation of ST3GAL4 expression and thereby reduces the adhesion of cancer cells to blood vessels, which results in inhibition of metastasis.

Topics: Animals; Antibodies; beta-Galactoside alpha-2,3-Sialyltransferase; Cell Adhesion; Cell Line, Tumor; Cell Movement; Down-Regulation; Female; Human Umbilical Vein Endothelial Cells; Humans; Kangai-1 Protein; Lewis X Antigen; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Oligonucleotide Array Sequence Analysis; Real-Time Polymerase Chain Reaction; RNA, Messenger; RNA, Small Interfering; Sialyl Lewis X Antigen; Sialyltransferases; Transplantation, Heterologous

Radical segmentectomy has been performed for small-sized non-small cell lung cancer (NSCLC). However, underestimation of mediastinal lymph node metastasis in the absence of hilar or interlobar metastasis (skip N2) affects surgical strategy. Our aim was to investigate preoperative and intraoperative predictors of skip N2 in clinical stage (c-stage) IA NSCLC.. From 1998 to 2011, 279 patients (155 men and 124 women) with c-stage IA NSCLC (230 pN0, 17 pN1, 12 skip N2, 20 non-skip N2) underwent systematic lobectomy (R0 resection) at our institute. We compared preoperative serum concentrations of carcinoembryonic antigen, cytokeratin 19 fragment, sialyl Lewis X (SLX), and pre- and intraoperative clinicopathological features of pN0 and skip N2 patients. Receiver operator characteristic (ROC) curve analysis was performed to distinguish between the two patient groups.. The 5-year survival rate of skip N2 patients was 78.6%, higher than that of non-skip N2 patients (44.9%), and not significantly different than that of pN0 (86.7%) or pN1 patients (82.4%). The mean serum SLX concentration in skip N2 patients (28.0 U/ml) was elevated compared to that in pN0 patients (22.9 U/ml). In ROC analysis of SLX, the area under the curve was 0.710, and the optimal cut-off value was 21.4 U/ml (sensitivity, 91.7%; specificity, 51.7%). In multivariate analysis, SLX was an independent predictor of skip N2 in patients with c-stage IA NSCLC (odds ratio, 9.43; p = 0.006).. Skip N2 metastasis is common in patients with c-stage IA NSCLC with high serum SLX, and lobectomy with complete dissection of hilar and mediastinal lymph nodes should remain the standard surgical procedure for these cases.

Topics: Aged; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Non-Small-Cell Lung; Female; Follow-Up Studies; Humans; Keratin-19; Lung Neoplasms; Male; Neoplasm Metastasis; Neoplasm Staging; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen; Survival Rate

We investigated various tumor markers in patients with surgically treated small cell lung cancer (SCLC) to identify the markers closely correlated to pathological staging and to predict survival by retrospective analyses.. Reviewing database records between 1990 and 2007 revealed 36 patients with SCLC, that were grouped according to clinical and pathological stages. Receiver operating characteristic (ROC) curves were calculated for serum levels of various tumor makers to predict the pathological stage. The cut-off value was calculated from the ROC curve of the significant marker. Survival in patient groups divided by the new cut-off value was calculated.. Serum levels of various tumor makers were not significantly different between the pathological stage groups, except for serum sialyl Lewis X (SLX). ROC curve of SLX was significantly correlated to pathological stages (P = 0.0136). The calculated SLX cut-off value was 25.1 U/ml, with 80% sensitivity and 70% specificity. Five-year survival of patients selected by this new cut-off was 82.5%, whereas that with the standard cut-off (38.0 U/ml) was 55.9%.. Serum SLX values were associated with pathological stage and survival after surgery in SCLC patients.

Topics: Aged; Aged, 80 and over; Biomarkers, Tumor; Female; Follow-Up Studies; Humans; Lung Neoplasms; Lymph Nodes; Male; Middle Aged; Neoplasm Staging; Oligosaccharides; Preoperative Care; Prognosis; Retrospective Studies; Sialyl Lewis X Antigen; Small Cell Lung Carcinoma; Survival Rate

The carbohydrate sialyl Lewis X (sLeX) is expressed on leukocytes and carcinoma cells and binds to selectins during inflammatory processes and early metastasis. Synthesis of sLeX depends on activity of enzymes, including α(1,3/1,4) fucosyltransferase (FucT-III). Tumor necrosis factor-α (TNF-α) up-regulates FucT-III, resulting in increased sLeX in the airways of patients with respiratory disease; however, the mechanisms that regulate sLeX in the inflammatory tumor microenvironment are not well understood.. The authors stably transfected human lung carcinoma cell lines with the FucT-III gene and exposed them to TNF-α to investigate its role in regulation of sLeX expression and selectin-binding ability using semiquantitative real-time polymerase chain reaction and flow cytometry. Cytokine expression was examined in transfected cells using chemiluminescent arrays and enzyme-linked immunosorbent assays, and invasion was studied using Matrigel assays and alterations in morphology. Human lung tissue arrays were analyzed for immunohistochemical detection of sLeX and neutrophils.. Stimulation of FucT-III-transfected cells with recombinant human (rh) TNF-α up-regulated sLeX expression and increased E-selectin binding. Transfected cells secreted high levels of interleukin 8, growth-regulated oncogene-α, and mast cell proteinase-1. Cells exposed to rhTNF-α, neutrophil-conditioned media, and cultures with a 5:1 ratio of neutrophils to cancer cells had significantly increased sLeX expression and invasiveness and underwent nonadherent morphologic changes. In lung carcinomas, but not in normal lung tissues, 71% of tumors were highly positive for sLeX expression in areas of increased neutrophil infiltration.. The current results indicated that neutrophils may be recruited to areas of FucT-III activity and sLeX expression in lung carcinomas to enhance the invasive and metastatic potential of lung cancer cells.

Topics: Blotting, Western; Carcinoma, Non-Small-Cell Lung; E-Selectin; Enzyme-Linked Immunosorbent Assay; Flow Cytometry; Fucosyltransferases; Humans; Immunoenzyme Techniques; Lung Neoplasms; Neoplasm Invasiveness; Neutrophils; Oligosaccharides; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sialyl Lewis X Antigen; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha

Interactions of endothelial selectins with tumour cell glycoconjugates have been shown to have a major role in tumour cell dissemination in previous experiments. However, experiments validating this observation were limited in value, as 'metastases' in these experiments were artificially induced by i.v. injection rather than developed spontaneously as in true metastases.. Endothelial (E) and platelet (P)-selectin-deficient severe combined immunodeficient (scid) mice were generated and human HT 29 colon cancer cells were subcutaneously inoculated in these mice and in wild-type scid mice. Tumour growth, spontaneous metastasis formation in the lung and adherence of HT29 cells to E- and P-selectin under flow were determined.. The number of metastases decreased by 84% in E- and P-selectin-deficient scid mice, compared with wild-type scid mice. The remaining 16% metastases in the E- and P-selectin-deficient scid mice grew within the pulmonary artery and not in the alveolar septae as they did in wild-type scid mice. Flow experiments indicate that tumour cells roll and tether on an E- and P-selectin matrix similar to leukocytes; however, firm adhesion is mainly mediated in E-selectin.. Our results indicate that E- and P-selectins have a crucial role in spontaneous metastasis formation. As the human HT 29 colon cancer cells are positive for the lectin Helix pomatia agglutinin (HPA), which identified the metastatic phenotype in earlier clinical studies, these results are of particular clinical relevance.

Topics: Animals; Cell Adhesion; Colonic Neoplasms; E-Selectin; HT29 Cells; Humans; Lectins; Lung Neoplasms; Mice; Mice, Inbred BALB C; Mice, SCID; Neoplastic Cells, Circulating; Oligosaccharides; P-Selectin; Sialyl Lewis X Antigen

Almost all of fucose and sialic acid in mucus are found on the mucus glycoproteins (mucins), and these sugar components on mucins are known to be associated with the viscous property of mucus. We have reported some aspects of carbocisteine, a mucoregulatory drug, correcting fucose and sialic acid contents in mucus. At present, carbocisteine's expectorant action of airway mucus is postulated to involve - the regulation of fucose and sialic acid contents on mucins. However little information is available about the relationship between the viscosity and sugar contents on mucins when treated with carbocisteine. To investigate further the mechanism behind the action of carbocisteine, the present study prepared MUC5AC fusion protein which has tandem repeat regions associated with MUC5AC, and evaluated the effects of carbocisteine on tumor necrosis factor (TNF)-alpha-induced increases of mucus viscosity and sialyl-Lewis x-epitopes antigen, an antigen which consists of fucosylated and sialylated sugar chains on the MUC5AC fusion proteins. Carbocisteine inhibited the TNF-alpha-induced increases of the viscosity and sialyl-Lewis x-epitopes on MUC5AC fusion protein. These findings suggest that carbocisteine may normalize the viscosity of mucus through "balancing" of fucose and sialic acid contents on airway mucins.

Topics: Carbocysteine; Carbohydrates; Cell Line, Tumor; Epitopes; Expectorants; Fucose; Humans; Lung Neoplasms; Mucins; Mucus; N-Acetylneuraminic Acid; Oligosaccharides; Recombinant Fusion Proteins; Respiratory System; Sialyl Lewis X Antigen; Tumor Necrosis Factor-alpha; Viscosity

Topics: Biomarkers, Tumor; Female; Humans; Lewis X Antigen; Lung Neoplasms; Male; Ovarian Neoplasms; Pancreatic Neoplasms; Sialyl Lewis X Antigen

Interactions between endothelial selectins and selectin ligands expressed on tumor cells have been implicated in the binding of circulating metastatic cancer cells to the vascular endothelium during extravasation. Moreover, there is mounting evidence that inflammatory environments can accelerate the progression of metastasis by neutrophil mediated mechanisms. In this study, a physiologically relevant in vivo model of early metastasis coupled with intravital microscopy was used to visualize the trafficking of tumor cells within the liver vasculature in real time. Using GFP-labeled Lewis lung carcinoma subline H-59 cells, we show here that disrupting the interactions between endothelial selectins and tumor cell selectin ligands diminished tumor cell recruitment to the liver. Furthermore, systemic inflammation induced by intravenous injection of lipopolysaccharide significantly enhanced the metastatic potential of these lung carcinoma cells by increasing their propensity to adhere to the liver sinusoidal endothelium. Confocal microscopy revealed frequent colocalization of cancer cells with neutrophils and neutrophil depletion in vivo significantly attenuated the lipopolysaccharide-induced increase in H-59 cell adhesion. Although direct selectin-selectin ligand interactions contributed significantly to tumor cell adhesion to sinusoidal endothelial cells, we show here that in addition, interactions between adherent neutrophils within the inflamed sinusoids and circulating tumor cells may further increase tumor cell arrest in the liver.

Topics: Animals; Carcinoma, Lewis Lung; Cell Adhesion; Disease Progression; Endothelium, Vascular; Inflammation; Ligands; Lipopolysaccharides; Liver; Liver Neoplasms; Lung Neoplasms; Mice; Mice, Inbred C57BL; Neutrophils; Oligosaccharides; Selectins; Sialyl Lewis X Antigen; Tumor Cells, Cultured

Topics: Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Humans; Keratin-19; Lung Neoplasms; Oligosaccharides; ROC Curve; Sialyl Lewis X Antigen

The metastatic breast cancer cell line, 4T1, abundantly expresses the oligosaccharide sialylated Lewis x (sLe(x)). SLe(x) oligosaccharide on tumor cells can be recognized by E- and P-selectin, contributing to tumor metastatic process. We observed that both selectins reacted with this cell line. However, contrary to the E-selectin reactivity, which was sLe(x) dependent, P-selectin reactivity with this cell line was sLe(x)-independent. The sLe(x)-Neg variant of the 4T1 cell line with markedly diminished expression of sLe(x) and lack of sLe(a), provided a unique opportunity to characterize P-selectin ligands and their contribution to metastasis in the absence of overlapping selectin ligands and E-selectin binding. We observed that P-selectin binding was Ca(2+)-independent and sulfation-dependent. We found that P-selectin reacted primarily with cell surface chondroitin sulfate (CS) proteoglycans, which were abundantly and stably expressed on the surface of the 4T1 cell line. P-selectin binding to the 4T1 cells was inhibited by heparin and CS glycosaminoglycans (GAGs). Moreover, Heparin administration significantly inhibited experimental lung metastasis. In addition, the data suggest that surface CS GAG chains were involved in P-selectin mediated adhesion of the 4T1 cells to murine platelets and human umbilical vein endothelial cells. The data suggest that CS GAGs are also the major P-selectin-reactive ligands on the surface of human MDA-MET cells. The results warrant conducting clinical studies on the involvement of cell surface CS chains in breast cancer metastasis and evaluation of various CS types and their biosynthetic pathways as target for development of treatment strategies for antimetastatic therapy of this disease.

Topics: Animals; Breast Neoplasms; Calcium; Cell Line, Tumor; Cell Membrane; Chondroitin Sulfates; Fucosyltransferases; Glycosaminoglycans; Heparin; Humans; Ligands; Lung Neoplasms; Mice; Mice, Inbred BALB C; Neoplasm Metastasis; Oligosaccharides; P-Selectin; Proteoglycans; Sialyl Lewis X Antigen; Transfection

This study aimed to establish the clinical significance of preoperative serum cytokeratin 19 fragment (CYFRA21-1) and sialyl-Lewis x (SLex) as prognostic markers.. The study involved 272 patients (181 male, 91 female; median age 69 years; range, 32 to 92) with non-small cell lung cancer (NSCLC) who underwent pulmonary resection with mediastinal lymph node dissection. Tumor markers carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), CYFRA21-1, and SLex were examined.. A log-rank test revealed that age, gender, performance status, CEA, SCC, CYFRA21-1, and SLex were associated with the survival rate. By multivariate analysis, age, gender, performance status, CYFRA21-1 (risk ratio, 2.42) and SLex (risk ratio, 6.18) were independent prognostic factors. For patients positive for both markers, the relative risk was 6.10 compared with patients negative for both markers. The patients were divided into three groups: negative for both CYFRA21-1 and SLex (n = 97); positive for either marker (n = 136); and positive for both markers (n = 39). The 1-, 3-, and 5-year survival rates were the following: 98%, 82%, and 75% in the first group; 90%, 63%, and 49% in the second group; and 62%, 31%, and 25% in the third group (p < 0.001). Sixty-four percent of patients positive for both markers were histologic stage III/IV, and 68% of patients negative for both markers were stage I.. Serum CYFRA21-1 and SLex were prognostic markers for NSCLC. Their combination should contribute to the classification of NSCLC patients. Preoperative staging should be carefully performed in patients positive for both tumor markers.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen

Topics: Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Humans; Keratin-19; Keratins; Lung Neoplasms; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen

This study aimed to establish the clinical significance of preoperative serum cytokeratin 19 fragment (CYFRA21-1) and Sialyl Lewis(x) (SLX) in patients with stage I non-small cell lung cancer (NSCLC). The study involved 137 patients (87 male, 50 female; median age 69 years) with completely resected stage I NSCLC. SLX, carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), and CYFRA21-1 were examined. Receiver operator characteristic (ROC) curves were constructed to determine prognostic cut-off values. Among the 137 patients, we identified 30 with recurrence within 3 years. The 5-year survival rates in patients with (n=30) and without (n=107) recurrence were 14% and 81%, respectively. The serum concentrations of SLX, CEA, and CYFRA21-1 in the recurrence group were significantly higher than those in the non-recurrence group. The areas under the ROC curve (AUC) were 0.72, 0.65, 0.53, and 0.64 for SLX, CEA, SCC, and CYFRA21-1, respectively. The prognostic cut-off values were 36U/ml, 7.8ng/ml, 1.5ng/ml, and 3.2ng/ml for SLX, CEA, SCC, and CYFRA21-1, respectively. A log-rank test revealed that age, performance status, T factor, lymphatic invasion, vascular invasion, SLX, CEA, SCC, and CYFRA21-1 were all significantly associated with survival. By multivariate analysis, age, performance status, lymphatic invasion, SLX (risk ratio, 4.11) and CYFRA21-1 (risk ratio, 3.47) were independent prognostic factors. For patients positive for both CYFRA21-1 and SLX, the relative risk was 5.32 compared with patients who were negative for both markers. The 5-year survival rates were 80% in the group negative for both markers (n=86); 52% in the group positive for one of the markers (n=43); and 13% for the group positive for both markers (n=8) (p<0.001). We concluded that serum SLX and CYFRA21-1 were prognostic markers for stage I NSCLC. Their combination should contribute to the classification of stage I NSCLC patients. There is a need to consider adjuvant and neoadjuvant therapies to improve prognosis in patients positive for both tumor markers.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Keratin-19; Keratins; Lung Neoplasms; Male; Middle Aged; Neoplasm Recurrence, Local; Neoplasm Staging; Oligosaccharides; Peptide Fragments; Sialyl Lewis X Antigen; Survival Rate; Time Factors; Treatment Outcome

Carbocisteine is a mucoregulatory drug normalizing sialic acid and fucose contents in mucins through the regulation of glycosyltransferase activities. Tumor necrosis factor (TNF)-alpha-induced overexpression of sialyl-Lewis x epitopes, containing sialic acid and fucose, in mucins were previously reported to be regulated by glycosyltransferase mRNAs expression through phosphatidyl inositol-specific phospholipase C (PI-PLC) signaling pathways [Ishibashi, Y., Inouye, Y., Okano, T., Taniguchi, A., 2005. Regulation of sialyl-Lewis x epitope expression by TNF-alpha and EGF in an airway carcinoma cell line. Glycoconj. J. 22, 53-62]. To investigate the mechanism behind the mucoregulatory action of carbocisteine, the present study evaluated the effects of carbocisteine on TNF-alpha-induced overexpression of sialyl-Lewis x epitopes in NCI-H292 cells. 100 mug/ml of carbocisteine was able to inhibit the TNF-alpha-induced expression of hST3GallV mRNA, FUT3 mRNA, C2/4GnT mRNA and sialyl-Lewis x epitopes as well as the TNF-alpha-induced activity of PI-PLC in NCI-H292 cells. These findings suggest that carbocisteine may normalize the sialyl-Lewis x epitopes expression in mucins through the inhibition of cellular PI-PLC activity in vivo.

Topics: Carbocysteine; Carcinoma; Cell Line, Tumor; Epitopes; Expectorants; Gene Expression Regulation, Enzymologic; Glycosyltransferases; Humans; Lung Neoplasms; Oligosaccharides; Phosphatidylinositol Diacylglycerol-Lyase; Phosphoinositide Phospholipase C; RNA, Messenger; Sialyl Lewis X Antigen; Tumor Necrosis Factor-alpha

The purpose of this study was to analyze the clinical significance of serum Sialyl Lewisx (SLX) concentrations as a predictor of N2 disease in patients with non-small-cell lung cancer.. The study included 272 patients with non-small-cell lung cancer who underwent pulmonary resection in our institution between January 1998 and December 2003. Of 272 patients, the serum concentrations of SLX were measured by using a commercially available radioimmunoassay kit.. The 5-year survival rates of patients with concentrations of SLX > 38 U/mL and those with lower concentrations were 32% and 69%, respectively (P < .0001). The median serum concentration of SLX in patients with multilevel N2 or N3, single-level N2, and N0/1 disease were 44, 30, and 27 U/mL, respectively. The concentrations of serum SLX in patients with multilevel N2 disease were significantly higher than those in patients with single-level N2 or those with N0/1 disease (Mann-Whitney U-test; P < .0001). Although the sensitivity of SLX for identifying patients with non-small-cell lung cancer was only 24% in all patients, the sensitivity of SLX increased as the N-factor increased; the sensitivity of N0/1 disease was 15%, that of single-level N2 disease was 22%, and that of multilevel N2 or N3 disease was 71%.. High serum concentrations of SLX predicted multilevel N2 disease and the associated poor outcome. Although the sensitivity of serum SLX is not acceptable for use as a screening tumor marker, we suggest that the serum concentration of SLX is useful as a staging marker to determine the strategy of treatment.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Carcinoma, Non-Small-Cell Lung; Carcinoma, Squamous Cell; Female; Humans; Lewis Blood Group Antigens; Lung Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Staging; Oligosaccharides; Prognosis; Retrospective Studies; Sialyl Lewis X Antigen; Survival Rate; Treatment Outcome

Sialyl-Lewis x epitopes and MUC5AC protein are known to be overexpressed in mucins secreted by patients suffering from various respiratory diseases. To investigate the mechanisms by which airway inflammatory agents mediate the expression of sialyl-Lewis x epitopes and MUC5AC mucin, we examined the effects of tumor necrosis factor (TNF)-alpha and epidermal growth factor (EGF) in the human lung carcinoma cell line, NCI-H292. Basal expression levels of hST3GalIV, FUT3 and C2/4GnT mRNA, involved in the biosynthesis of sialyl-Lewis x, were higher than those of other glycosyltransferases in NCI-H292 cells. TNF-alpha induced expression of hST3GalIV, FUT3, C2/4GnT and MUC5AC mRNAs in NCI-H292 cells. When cells were pretreated with U73122, a phosphatidylinositol-phospholipase C (PI-PLC) inhibitor, the expression of these glycosyltransferase mRNAs was suppressed. Treating cells with EGF induced the down-regulation of these glycosyltransferase mRNAs and sialyl-Lewis x epitopes, while inducing an increase in expression of MUC5AC mRNA. These EGF-mediated effects on the glycosyltransferase and MUC5AC mRNAs were blocked when cells were first exposed to AG1478, an EGF receptor tyrosine kinase inhibitor. These findings suggest that the expression of sialyl-Lewis x epitopes, which is regulated separately from the expression of MUC5AC protein, may be controlled through pathways such as the EGF receptor tyrosine kinase and PI-PLC signaling cascades in NCI-H292 cells.

Topics: Cell Line, Tumor; Epidermal Growth Factor; Epitopes; ErbB Receptors; Gene Expression Regulation; Humans; Lung Neoplasms; Mucin 5AC; Mucins; Oligosaccharides; Phosphatidylinositol Diacylglycerol-Lyase; Phosphoinositide Phospholipase C; RNA, Messenger; Sialyl Lewis X Antigen; Tumor Necrosis Factor-alpha

Expression of sialyl Lewis(x) (sLe(x)) and sLe(a) on tumor cells is thought to facilitate metastasis by promoting cell adhesion to selectins on vascular endothelial cells. Experiments supporting this concept usually bypass the early steps of the metastatic process by employing tumor cells that are injected directly into the blood. We investigated the relative role of sLe(x) oligosaccharide in the dissemination of breast carcinoma, employing a spontaneous murine metastasis model. An sLe(x) deficient subpopulation of the 4T1 mammary carcinoma cell line was produced by negative selection using the sLe(x)-reactive KM93 MAb. This subpopulation was negative for E-selectin binding but retained P-selectin binding. Both sLe(x)-negative and -positive cells grew at the same rate; however, sLe(x)-negative cells spread more efficiently on plates and had greater motility in wound-scratch assays. Mice inoculated in the mammary fat pad with sLe(x)-negative and -positive variants produced lung metastases. However, the number of lung metastases was significantly increased in the group inoculated with the sLe(x)-negative variant (p = 0.0031), indicating that negative selection for the sLe(x) epitope resulted in enrichment for a subpopulation of cells with a high metastatic phenotype. Cell variants demonstrated significant differences in cellular morphology and pattern of tumor growth in primary and secondary tumor sites. These results strongly suggest that loss of sLe(x) may facilitate the metastatic process by contributing to escape from the primary tumor mass.

Topics: Animals; E-Selectin; Female; Ligands; Lung Neoplasms; Mammary Neoplasms, Experimental; Mice; Mice, Inbred BALB C; Neoplasm Metastasis; Oligosaccharides; Sialyl Lewis X Antigen

Clustered presentation of sialyl Lewis X (sLe(X)) on tumor cell mucins is thought to facilitate metastasis through binding to selectin adhesion receptors expressed on platelets, leukocytes and endothelial cells. Thus, interfering with sLe(X) assembly might provide a chemotherapeutic method for treating metastatic disease. Prior studies have shown that peracetylated disaccharides can act in cells as substrates for the assembly of oligosaccharides related to sLe(X) synthesis, and the assembly of oligosaccharides on the disaccharides diverts the assembly of sLe(X) from endogenous cell surface glycoconjugates. Here, we show that treatment of cultured human adenocarcinoma cells with micromolar concentrations of the peracetylated disaccharide, (Ac)(6)GlcNAcbeta1,3Galbeta-O-naphthalenemethanol (AcGnG-NM) reduces the expression of sLe(X) and diminishes binding in vitro to selectin-coated dishes, thrombin-activated platelets, and tumor necrosis factor alpha-activated endothelial cells. Altering glycosylation in this way significantly reduced the ability of tumor cells to distribute to the lungs of wild-type mice over a 3-h period after i.v. injection. No significant difference in biodistribution was noted after the injection of AcGnG-NM-treated tumor cells into P-selectin deficient mice, although the extent of lung seeding was reduced compared with that in wild-type mice. In vitro, we demonstrate that normal mouse platelets, but not P-selectin-deficient platelets, bound to control tumor cells and protected them from leukocyte-mediated cytolysis. Conversely, treatment of tumor cells with disaccharide markedly reduced the ability of normal platelets to protect them from cytolysis. Finally, in an experimental metastasis model, we show that treatment of tumor cells with the disaccharide markedly reduced their lung colonization potential after injection into severe combined immunodeficient mice. These findings suggest that this compound may represent a novel class of chemotherapeutic agents for prevention and treatment of metastatic disease.

Topics: Adenocarcinoma; Blood Platelets; CA-19-9 Antigen; Carbohydrate Sequence; Cell Adhesion; Colonic Neoplasms; Disaccharides; Endothelium, Vascular; Gangliosides; Glycosylation; Humans; Lung Neoplasms; Molecular Sequence Data; Neoplasm Metastasis; Oligosaccharides; Sialyl Lewis X Antigen; Tissue Distribution; Tumor Cells, Cultured

Endothelial carbohydrate binding proteins, E- and P-selectins, are thought to mediate sialyl Lewis A/X-dependent hematogenous cancer metastasis. We tested this hypothesis using sialyl Lewis X-dependent B16 melanoma lung targeting and its inhibition with selectin ligand mimicry peptide, IELLQAR. In E/P-selectin doubly deficient mutant mice, sialyl Lewis X-expressing B16 melanoma cells colonized the lung, and IELLQAR inhibited this colonization. However, tumors grown in E/P-selectin-deficient mice were significantly smaller than those grown in wild-type mice. These results indicate that the IELLQAR peptide receptor expressed in the lung vasculature plays a major role in sialyl Lewis X-dependent cancer cells targeting to the lung.

Topics: Animals; E-Selectin; Lung; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Mice, Mutant Strains; Molecular Mimicry; Neovascularization, Pathologic; Oligopeptides; Oligosaccharides; P-Selectin; Receptors, Peptide; Sialyl Lewis X Antigen

Epithelial carcinoma and leukemia cells express sialyl Lewis x oligosaccharides as tumor-associated carbohydrate antigens. To determine the role of sialyl Lewis x oligosaccharides in tumor dissemination, human melanoma MeWo cells, which do not express sialyl Lewis x, were transfected with alpha1,3-fucosyltransferase III (FTIII), and cell lines expressing different amounts of sialyl Lewis x were isolated. When these cells were injected into the tail vein of nude mice, cells expressing moderate amounts of sialyl Lewis x (MeWo-FTIII.M) produced a significantly greater number of lung tumor foci than did parental MeWo cells. In contrast, cells expressing large amounts of sialyl Lewis x (MeWo-FTIII.H) produced few lung tumor foci in nude mice but were highly tumorigenic in beige mice, which have defective natural killer (NK) cells. In vitro assays demonstrated that MeWo-FTIII.H cells are much more sensitive to NK cell-mediated cytotoxicity than are MeWo-FTIII.M cells or parental MeWo cells and the susceptibility of MeWo-FTIII.H cells to NK cell-mediated cytolysis can be inhibited by preincubating MeWo-FTIII.H cells with anti-sialyl Lewis x antibody. Moreover, we discovered that NK cell-mediated cytolysis of MeWo-FTIII.H cells can be inhibited by the addition of an antibody against the NK cell receptor CD94 or sialyl Lewis x oligosaccharides. These results, combined with structural analysis of MeWo-FTIII.H cell carbohydrates, indicate that moderate amounts of sialyl Lewis x lead to tumor metastasis, whereas expression of high levels of sialyl Lewis x leads to an NK cell attack on tumor cells, demonstrating that expression of different amounts of sialyl Lewis x results in entirely different biological consequences.

Topics: Animals; Antigens, CD; Cytotoxicity, Immunologic; Female; Fucosyltransferases; Gene Expression; Humans; Killer Cells, Natural; Lectins, C-Type; Lung Neoplasms; Mice; Mice, Nude; Mice, SCID; Neoplasms, Experimental; NK Cell Lectin-Like Receptor Subfamily D; Oligosaccharides; Sialyl Lewis X Antigen; Transfection; Tumor Cells, Cultured

To evaluate the prognostic relevance of CEACAM1 and sialyl Lewis X expression in adenocarcinomas of the lung.. Paraffin wax sections of 93 patients with adenocarcinomas of the lung who underwent surgery between 1990 and 1995 were immunohistochemically investigated using monoclonal anti-CEACAM1 and sialyl Lewis X antibodies. The clinical course of all patients was followed up for a minimum of 5 years.. Sixty-one tumors were classified as CEACAM1-positive, and 32 were classified as CEACAM1-negative. Patients with CEACAM1-positive tumors had a significantly poorer overall (P =.00025) and relapse-free (P =.00029) survival than those with CEACAM1-negative tumors. Only three patients did not express the sialyl Lewis X glycotope, whereas 90 tumors (97%) were sialyl Lewis X-positive. In multivariate Cox regression analysis, next to tumor stage and sex, only the expression of CEACAM1 was a significant independent prognostic factor for survival.. Expression of CEACAM1 was an independent prognostic factor in our patient population and can be used to stratify patients with adenocarcinomas of the lung into low-risk and high-risk groups. In contrast, the expression of sialyl Lewis X was of no prognostic relevance because it was expressed in 97% of all investigated tumors, and most likely has no influence on the function of CEACAM1 in this tumor entity.

Topics: Adenocarcinoma; Adult; Aged; Antibodies, Monoclonal; Antigens, CD; Antigens, Differentiation; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Cell Adhesion Molecules; Female; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Lewis Blood Group Antigens; Lewis X Antigen; Lung Neoplasms; Male; Middle Aged; Oligosaccharides; Prognosis; Retrospective Studies; Risk Factors; Sialyl Lewis X Antigen; Survival Analysis

Sialidase expression levels are inversely correlated with the metastatic potential of mouse colon adenocarcinoma 26 sublines, as assessed by activity assays and RT-PCR, irrespective of total and cell surface sialic acid contents. Compared with low metastatic NL4 and NL44 cell lines, the highly metastatic NL17 and NL22 cells exhibit low expression of sialidases, accompanied with higher levels of sialylLe(x) and GM3. To investigate whether these properties of NL17 cells can be altered by sialidase overexpression, we transfected a cytosolic sialidase gene into NL17 cells. The result was markedly inhibited lung metastasis, invasion and cell motility with a concomitant decrease in sialylLe(x) and GM3 levels, in line with the case of spontaneously low metastatic sublines having relatively high endogenous sialidase levels, implying that sialidase level is a determining factor affecting metastatic ability. Treatment of the cells with antibodies against sialylLe(x) and GM3 affected cell adhesion and/or cell motility, providing evidence that desialylation of these molecules, as targets of sialidase, is involved in the suppression of metastasis.

Topics: Adenocarcinoma; Animals; Cell Line; Chromatography, Thin Layer; Colonic Neoplasms; DNA Primers; Female; Flow Cytometry; G(M3) Ganglioside; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Inbred C57BL; Mice, Nude; Oligosaccharides; Reverse Transcriptase Polymerase Chain Reaction; Sialyl Lewis X Antigen; Sialyltransferases; Transfection; Tumor Cells, Cultured

Sialyl Lewis X-i antigen has been used as a diagnostic tool for lung adenocarcinoma. However, serum levels of the antigen are also raised in some patients with idiopathic pulmonary fibrosis (IPF) without coexistent malignancy. Expression of the antigen in serum samples of patients with lung adenocarcinoma was evaluated and compared with that of patients with IPF by Western blotting in order to establish a specific laboratory test to differentiate lung adenocarcinoma from IPF.. The pattern of antigen expression in serum samples from 23 patients with either lung adenocarcinoma or non-malignant lung disease in whom serum levels of sialyl Lewis X-i antigen (>50 U/ml) were significantly increased was studied by Western blotting.. Thirteen of the 14 serum samples from patients with lung adenocarcinoma had a molecular weight band at 120 or 130 kD, while five of the six patients with IPF had two or three bands at <97.4 kD. The pattern of antigen expression was apparently different between the two diseases. The sensitivity, specificity, positive likelihood ratio, and negative likelihood ratio of this test in 20 patients with lung adenocarcinoma and IPF were 92.9%, 83.3%, 5.57, and 0.09, respectively.. Western blotting analysis of serum samples from patients with raised levels of sialyl Lewis X-i antigen provides some clinical information for a differential diagnosis between lung adenocarcinoma and IPF.

Topics: Adenocarcinoma; Adult; Aged; Biomarkers, Tumor; Blotting, Western; Female; Humans; Lung Neoplasms; Male; Middle Aged; Oligosaccharides; Pulmonary Fibrosis; Sialyl Lewis X Antigen

Carcinoembryonic antigen (CEA), cancer antigen 125 (CA 125), NCC-ST-439, carbohydrate antigen 19-9 (CA 19-9), cytokeratin 19 fragment (CYFRA 21-1), sialyl Lewis X-i antigen (SLX), progastrin-releasing peptide (ProGRP), squamous cell carcinoma antigen (SCC) and neuron specific enolase (NSE) were evaluated in the pleural effusion of 39 patients with lung cancer (29 adenocarcinomas, seven small-cell carcinomas, three squamous cell carcinomas) and 43 patients with tuberculous pleurisy. The levels of the tumor markers other than SCC and NSE were significantly higher in lung cancer than in tuberculosis. High levels of CYFRA 21-1 and SCC were observed in squamous cell carcinoma and high levels of ProGRP and NSE were observed in small-cell carcinoma. According to the validity score, sensitivity (%) + specificity (%) - 100, the optimal cut-off levels of pleural effusion were 8.1 ng/ml for CEA, 660 U/ml for CA 125, 2.6 U/ml for NCC-ST-439, 10 U/ml for CA 19-9, 65 ng/ml for CYFRA 21-1, 140 U/ml for SLX, 23.2 pg/ml for ProGRP, 0.6 ng/ml for SCC and 5 ng/ml for NSE. By comparison of validity scores for each optimal cut-off level and of receiver operating characteristic (ROC) curves, we suggest that a CEA assay is the most useful for pleural effusion. The combined assay of CEA + ProGRP and CEA + ProGRP + CYFRA 21-1 were considered to be useful.

Topics: Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; CA-125 Antigen; CA-19-9 Antigen; Carcinoembryonic Antigen; Humans; Keratin-19; Keratins; Lung Neoplasms; Oligosaccharides; Peptide Fragments; Peptides; Phosphopyruvate Hydratase; Pleural Effusion, Malignant; Recombinant Proteins; Serpins; Sialyl Lewis X Antigen; Tuberculosis, Pleural

Inflammatory breast carcinoma (IBC) is characterized by florid tumor emboli within lymphovascular spaces termed lymphovascular invasion (LVI). Using a human-scid model of IBC (MARY-X), we have demonstrated using retrovirally-mediated dominant-negative E-cadherin mutant approaches (H-2K(d)-E-cad), that the tumor cell embolus (IBC spheroid) forms on the basis of an intact and overexpressed E-cadherin/alpha, beta-catenin axis which mediates tumor cell-tumor cell adhesion analogous to the embryonic blastocyst and accounts for the compactness of the embolus. The tumor cell embolus (IBC spheroid), in contrast, fails to bind the surrounding vascular endothelial cells both in vitro and in vivo because of markedly decreased sialyl-Lewis X/A carbohydrate ligand-binding epitopes on its overexpressed MUC1 which are necessary for binding endothelial cell E-selectin. This tumor cell-endothelial cell aversion further contributes to the compactness of the IBC spheroid and its passivity in metastasis dissemination. This passivity is manifested by a dramatic increase in metastatic pulmonary emboli following palpation of the primary tumor. In assessing this passivity of metastatic dissemination, we compared the effects of palpation on MARY-X with the effects of palpation on a derived dominant-negative E-cadherin mutant (H-2K(d)-E-cad), as well as other well known human tumoral xenografts exhibiting no (MCF-7, T47D), low (MDA-MB-231, MDA-MB-468) or high (C8161, M24(met)) levels of spontaneous metastasis but no LVI. Palpation of each xenograft similarly increased intratumoral pressure by 200% (10-->30 mmHg) but dramatically increased the numbers and sizes of pulmonary metastases 10-100-fold (P<0.001) only in MARY-X. The mechanism of this effect was through an immediate post-palpation release of circulating tumor emboli detected 2-3 min after palpation (P<0.01) by human cytokeratin 19 RT-PCR of extracted RNA from 300 microl of murine blood. Although circulating human tumor cell-derived growth factors (IGF-I, IGF-II, TGF-alpha and TGF-beta) and angiogenic factors (VEGF and bFGF) were detected by ELISA in murine serum of MARY-X, palpation did not further increase the circulating levels of these factors (P>0.1). Our findings support the cooperative role of E-cadherin and sialyl-Lewis X/A-deficient MUC1 in the passive dissemination of tumor emboli in IBC.

Topics: Adenocarcinoma; Animals; Antigens, Tumor-Associated, Carbohydrate; Cadherins; Cell Adhesion; Cells, Cultured; Endothelium, Vascular; Female; Humans; Lewis Blood Group Antigens; Lung Neoplasms; Mammary Neoplasms, Experimental; Mice; Mice, SCID; Mucin-1; Mutation; Neoplastic Cells, Circulating; Oligosaccharides; Sialyl Lewis X Antigen; Tumor Cells, Cultured

Topics: Animals; Carbohydrate Metabolism; Carcinoma, Lewis Lung; Cell Adhesion; Cells, Cultured; CHO Cells; Cricetinae; E-Selectin; Endothelium, Vascular; Humans; Intercellular Adhesion Molecule-1; Ligands; Liposomes; Lung Neoplasms; Neoplasm Metastasis; Oligosaccharides; Phosphatidylethanolamines; Phosphorylcholine; Polyethylene Glycols; Sialyl Lewis X Antigen; Tumor Cells, Cultured

Cell surface carbohydrates of epithelial cells play important roles in tumor progression. Previously, we have shown that expression of core 2 branched O-glycans in colorectal cancer is closely correlated with the vessel invasion and depth of invasion (K. Shimodaira et al., Cancer Res., 57: 5201-5206, 1997). To test whether this is also the case in human lung cancer, we have examined the expression pattern of core 2 beta1,6-N-acetylglucosaminyltransferase (C2GnT) mRNA responsible for the biosynthesis of core 2 branched O-glycans in 41 cases of lung cancer. Using in situ hybridization, C2GnT mRNA was detected in 73.2% of the lung cancer cells, irrespective of the histopathological type; whereas in normal lung tissues, its expression was restricted to the basal cells of bronchial mucosa. These results indicate that the expression level of C2GnT mRNA was significantly enhanced in association with malignant transformation. Statistical analysis between the C2GnT mRNA expressed in pulmonary adenocarcinoma and clinicopathological variables revealed that the expression of C2GnT was correlated with vessel invasion and lymph node metastasis with significant difference (P < 0.05), but expression of sialyl Le(x), which is frequently expressed in the adenocarcinoma, was not significantly correlated with lymph node metastasis. These results indicate that C2GnT mRNA detected by in situ hybridization reflects the malignant potentials of pulmonary adenocarcinoma, because lymph node metastasis is the most affecting factor to the patients' prognosis.

Topics: Adenocarcinoma; CA-19-9 Antigen; Carbohydrate Sequence; Gangliosides; Humans; In Situ Hybridization; Lung Neoplasms; Lymphatic Metastasis; Molecular Sequence Data; N-Acetylglucosaminyltransferases; Neovascularization, Pathologic; Oligosaccharides; RNA, Messenger; Sialyl Lewis X Antigen

Human lung tumor alpha1,3/4-L-fucosyltransferase (FT) was purified (2000-fold, 29% recovery) from 290 g of tissue by including a chromatography step on Affinity Gel-GDP. Two molecular forms (FTA, larger size carrying 15% alpha1,4-FT activity; FTB, the major form with 85% activity) were separated by further fractionation on a Sephacryl S-100 HR column. A difference in the electrophoretic mobilities of these two activities was also found on native polyacrylamide gel electrophoresis (PAGE). Both forms were devoid of typical alpha1,2-fucosylating activity but were associated with the novel alpha1,2-fucosylating ability of converting the Lewis a determinant to Lewis b. Based on percentage activity toward 2-O-MeGalbeta1,3GlcNAcbeta-O-Bn, both forms exhibited the same extent of activity toward various acceptors, which included sulfated, sialylated, or methylated LacNAc type 1 or type 2 as well as mucin core 2 acceptors. However, FTA and FTB exhibited a difference in their ability to act on mucin core 2 3'-sialyl LacNAc (activities 24.2% and 40.8%, respectively, as compared to 2-O-MeGalbeta1,3GlcNAcbeta-O-Bn). The unsubstituted LacNAc type 1 acceptors were 15-20 times as active as the corresponding LacNAc type 2 acceptors. The 3-O-substitution on the beta1,4-linked Gal (methyl, sulfate, or sialyl) in mucin core 2 acceptors increased the efficiency of these acceptors five- to eightfold. The most efficient acceptor for FTA and FTB was 3-O-sulfoGalbeta1,3GlcNAcbeta-O-Al (K(m) 100 and 47 microM, respectively). The K(m) (mM) values for 2-O-methyl Galbeta1,3GlcNAcbeta-O-Bn and 3-O-sialyl Galbeta1,3GlcNAcbeta-O-Bn were 0.40 and 2.5 (FTA) and 0.16 and 0.67 (FTB), respectively. The 35-kDa glycoprotein ancrod (from Malayan pit viper venom) containing 36% complex N-glycans with the antennae NeuAcalpha2,3Galbeta1,3GlcNAcbeta- acted as the best macromolecular acceptor substrate (K(m): 45 microM), as examined with FTB. On desialylation the acceptor efficiency dropped to approximately 50% (K(m) for asialo ancrod: 167 microM). Sialylglycoproteins, such as carcinoembryonic antigen, fetuin, and bovine alpha(1)-acid glycoprotein, were better acceptors than asialo fetuin. On the contrary, fetuin triantennary glycopeptide containing predominantly NeuAcalpha2,3Galbeta1,4GlcNAcbeta- was only 55% active as compared to the asialo glycopeptide (K(m): 1.43 and 0.63 mM, respectively). Thus, the human lung tumor alpha1,3/4-L-FT has the potential to generate clustered sialyl Lewis a and Lewis

Topics: Adenocarcinoma, Mucinous; Antigens, Neoplasm; Carbohydrate Sequence; Electrophoresis, Polyacrylamide Gel; Female; Fucosyltransferases; Glycopeptides; Glycoproteins; Humans; In Vitro Techniques; Lung Neoplasms; Middle Aged; Molecular Sequence Data; Mucins; Oligosaccharides; Sialyl Lewis X Antigen; Substrate Specificity

Selectins bind to carbohydrate ligands in a calcium-dependent manner and play critical roles in host defense and possibly in tumor metastasis. To isolate peptides that mimic E-selectin ligands, we screened a phage peptide library using E-selectin as a target molecule. This attempt unexpectedly failed, probably because the binding affinity of E-selectin to its ligand is low. We then took an approach that is analogous to the isolation of anti-idiotype antibodies and were able to isolate peptides that bound to anticarbohydrate antibodies recognizing E-selectin ligands. These peptides, enriched for their binding to anti-Lewis A antibody, were found to bind to E-, P- and L-selectins in a calcium-dependent manner. Phage harboring the identified peptide IELLQAR and synthetic peptides having the same sequence inhibited the binding of sialyl Lewis X or sialyl Lewis A oligosaccharides to E-selectin. The adhesion of HL-60 and B16 melanoma cells expressing sialyl Lewis X to E-selectin was also inhibited by the phage-displaying IELLQAR peptide. Moreover, i.v. injected IELLQAR peptide inhibited the lung colonization of mouse B16 melanoma and human lung tumor cells expressing sialyl Lewis X. These results demonstrate that it is possible to isolate peptides mimicking carbohydrate ligands by screening the peptides for binding to anticarbohydrate antibodies and then using them to inhibit carbohydrate-dependent experimental tumor metastasis.

Topics: Animals; Brevican; Cell Adhesion; Chondroitin Sulfate Proteoglycans; E-Selectin; HL-60 Cells; Humans; Lectins, C-Type; Lewis Blood Group Antigens; Lung; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Nerve Tissue Proteins; Oligopeptides; Oligosaccharides; Peptide Fragments; Recombinant Fusion Proteins; Sialyl Lewis X Antigen; Transfection

Carbohydrates on tumor cells have been shown to play an important role in tumor metastasis. We demonstrated before that CD24, a Mr 35,000-60,000 mucine-type glycosylphosphatidylinositol-linked cell surface molecule, can function as ligand for P-selectin and that the sialylLex carbohydrate is essential for CD24-mediated rolling of tumor cells on P-selectin. To investigate the role of both antigens more closely, we transfected human A125 adenocarcinoma cells with CD24 and/or fucosyltransferase VII (Fuc TVII) cDNAs. Stable transfectants expressed CD24 and/or sialylLex. Biochemical analysis confirmed that in A125-CD24/FucTVII double transfectants, CD24 was modified with sialylLex. Only double transfectants showed rolling on P-selectin in vivo. When injected into mice, double transfectants arrested in the lungs, and this step was P-selectin dependent because it was strongly enhanced in lipopolysaccharide (LPS) pretreated wild-type mice but not in P-selectin knockout mice. CD24 modified by sialylLex was required on the tumor cells because the LPS-induced lung arrest was abolished by removal of CD24 from the cell surface by phosphatidylinositol-specific phospholipase C. A125-FucTVII single transfectants expressing sialylLex but not CD24 did not show P-selectin-mediated lung arrest. The sialylLex epitope is abundantly expressed on human carcinomas, and significant correlations between sialylLex expression and clinical prognosis exist. Our data suggest an important role for sialylLex-modified CD24 in the lung colonization of human tumors.

Topics: Adenocarcinoma; Animals; Antigens, CD; Blood Platelets; CD24 Antigen; Cell Adhesion; Cell Movement; Female; Fucosyltransferases; Glycosylation; Humans; Lung; Lung Neoplasms; Male; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Oligosaccharides; P-Selectin; Phosphatidylinositol Diacylglycerol-Lyase; Phosphoinositide Phospholipase C; Sialyl Lewis X Antigen; Signal Transduction; Transfection; Type C Phospholipases

Lewis X, Sialyl Lewis X and Sialyl Dimeric Lewis X are three fucosylated glycoconjugates on cell surface. With immunohistochemical method, the expression of the three structures in the original lung cancer tissues (with or without metastasis), adjacent tissues and metastatic lesions of lung carcinoma were studied. It was found that the three antigens were expressed with different intensity on the cell surface and in the cytoplasm of the lung carcinoma cell. However, there was no or only trace expression of these antigens in the adjacent tissues of lung carcinoma and normal lung tissues. Moreover, the original lesions of lung carcinoma with metastasis and/or poor differentiation expressed higher level of the three antigens than those without metastasis and/or with well or medium differentiation. Sialyl Lewis X was considered to be more closely related to the metastatic ability and differentiation of the lung carcinoma cell than the other two antigens, Lewis X and Sialyl Dimeric Lewis X. Furthermore, in the lymph nodes with lung carcinoma cell metastasis, there were expression of the three antigens with different degree, while in those lymph nodes without lung carcinoma cell metastasis, there was no expression of the three antigens.

Topics: Glycoconjugates; Humans; Immunohistochemistry; Lewis X Antigen; Lung Neoplasms; Lymph Nodes; Lymphatic Metastasis; Oligosaccharides; Sialyl Lewis X Antigen

Monoclonal antibodies and lectins were used to examine the expression patterns of apical membrane oligosaccharide sequences specific to type II pneumocytes in atypical adenomatous hyperplasia (AAH) and lung cancer. Atypical cells of AAH and papillary adenocarcinoma cells expressed abundant sialyl Thomsen-Friedenreich (TF) antigen: this was not observed in acinar adenocarcinoma, bronchioloalveolar carcinoma with mucin production or squamous cell carcinoma. Sialyl Tn antigens was also detected on a few cells in AAH and papillary adenocarcinomas. Asialo TF and Tn antigen were not observed on the surface of carcinoma cells of any type. Alpha(alpha)2,3-linked sialic acids predominated in type II pneumocyte, AAH and papillary adenocarcinoma, whereas ciliated columnar cells expressed alpha2,6-linked sialic acids. Lewisx and sialyl Lewisx antigens capped the TF antigen in both O- and N-linked side chains on the surface of AAH and papillary adenocarcinoma cells, but were not expressed by type II pneumocytes. The findings demonstrate that papillary adenocarcinoma cells resemble type II pneumocytes in that they express abundant sialyl TF surface antigen, but they also express TF-related antigens not found in type II pneumocytes. Apical surface glycoconjugates of AAH have structural characteristics shared by both type II pneumocytes and papillary adenocarcinoma cells.

Topics: Adenocarcinoma; Antigens, Tumor-Associated, Carbohydrate; Glycoconjugates; Humans; Lung; Lung Neoplasms; Oligosaccharides; Sialyl Lewis X Antigen

Aberrant expression of cell surface carbohydrates such as sialyl Lewis X is associated with tumor formation and metastasis. In order to determine the roles of sialyl Lewis X in tumor metastasis, mouse melanoma B16-F1 cells were stably transfected with alpha1, 3-fucosyltransferase III to express sialyl Lewis X structures. The transfected B16-F1 cells, B16-FTIII, were separated by cell sorting into three different groups based on the expression levels of sialyl Lewis X. When these transfected cells were injected into tail veins of C57BL/6 mice, B16-FTIII.M cells expressing moderate amounts of sialyl Lewis X in poly-N-acetyllactosamines produced large numbers of lung tumor nodules. Surprisingly, B16-FTIII.H cells expressing the highest amount of sialyl Lewis X in shorter N-glycans died in lung blood vessels, producing as few lung nodules as B16-FTIII.N cells which lack sialyl Lewis X. In contrast, B16-FIII.H cells formed more tumors in beige mice and NK cell-depleted C57BL/6 mice than did B16-FTIII.M cells. B16-FTIII.H cells bound to E-selectin better than did B16-FTIII.M cells, but both cells grew at the same rate. These results indicate that excessive expression of sialyl Lewis X in tumor cells leads to rejection by NK cells rather than tumor formation facilitated by attachment to endothelial cells.

Topics: Animals; Cell Division; Cytotoxicity, Immunologic; E-Selectin; Flow Cytometry; Fucosyltransferases; Graft Rejection; Killer Cells, Natural; Lewis X Antigen; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Neoplasm Metastasis; Neoplasm Transplantation; Oligosaccharides; Recombinant Proteins; Sialyl Lewis X Antigen; Transfection; Tumor Cells, Cultured

E-Selectin is an inducible adhesion molecule, which is expressed on cytokine-activated endothelial cells and is thought to interact with cancer cells to initiate metastases. The relationship between serum E-selectin levels and prognoses in 101 patients with resected non-small cell lung cancers (NSCLCs) was studied, and survival curves were compared in relation to E-selectin levels and expression of two carbohydrate antigens, Sialyl Lewisx (SLX) and Sialyl Lewisa (CA19-9), which were immunohistochemically detected in resected specimens in 65 of the 101 cases. The serum E-selectin level on admission was 48.9+/-25.7 ng/ml (mean+/-SD, n=101), and the E-selectin-positive rate was 22.7%, being correlated with the progression of T-factor. The high E-selectin group showed a significantly worse survival rate than the normal E-selectin group. Multivariate analysis confirmed the significant prognostic value of E-selectin. The mean postoperative E-selectin level in 52 cases (36.93 ng/ml) was significantly lower than the preoperative E-selectin level (43.57 ng/ml), indicating that certain NSCLCs might induce the expression of E-selectin. In cases expressing carbohydrate antigens (SLX, CA19-9), the high E-selectin group showed a significantly worse survival curve than the normal E-selectin group. On the other hand, there was no significant difference in the survival curve between the high and normal E-selectin groups when carbohydrate antigens were negative. These results suggest that patients who have high serum E-selectin levels, especially with carbohydrate antigen-positive NSCLC, might be expected to have poor prognoses.

Topics: Adult; Aged; Aged, 80 and over; Analysis of Variance; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Carcinoma, Non-Small-Cell Lung; Cell Adhesion Molecules; E-Selectin; Female; Humans; Immunohistochemistry; Lung Neoplasms; Male; Middle Aged; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen; Solubility; Survival Rate; Time Factors

The morphology, cell growth, antigenic expression and tumorigenicity of cell subpopulations from the A549 lung adenocarcinoma isolated by Percoll gradient separation have been analysed. Four subpopulations were obtained (subpopulations A, B, C and D). Immunocytochemical analysis of several antigens was performed with monoclonal antibodies (MAbs): MUC1 mucin (C595, HMFG1 and HMFG2), MUC5B (PANH2); gp230 (PANH4); carbohydrate antigens including sialyl Lewis x (KM93), Tn antigen (83D4), Lewis y (C14); 5, 6, 8, 17 and 19 cytokeratins and p53. The cell population D tended to form cell aggregates that piled up on the monolayer similar to overgrowth cultures of the A549 parental cell line, whereas A, B and C cell subpopulations formed well spread monolayers. Both parental A549 and subpopulation D secreted abundant mucus. The topographic distribution and secretion production were correlated with tumorigenic assays since only subpopulation D grew in nude mice exhibiting reduced latency period; these characteristics correlated with the fast growth of the subpopulation D in vitro. Immunocytochemical analysis demonstrated that subpopulation D showed greater expression of MUC1 mucin and carbohydrate antigens such as Tn antigen, sialyl Lewis x and Lewis y and less expression of cytokeratins, p53, MUC5B and gp230; conversely, subpopulations A, B and C showed the opposite antigenic profile. Our results illustrate heterogeneity in the A549 cell line; subpopulations A, B and C retained characteristics of more differentiated adenocarcinoma while subpopulation D displayed features of a less differentiated tumor line.

Topics: Adenocarcinoma; Animals; Antibodies, Monoclonal; Cell Separation; Humans; Immunohistochemistry; Keratins; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Transplantation; Oligosaccharides; Phenotype; Sialyl Lewis X Antigen; Time Factors; Tumor Cells, Cultured; Tumor Suppressor Protein p53

Changes in cell membrane carbohydrate antigens play an important role in metastatic potential associated with carcinogenesis and in prognostic factors. We investigated immunohistochemically the expression of CD15 and sialyl CD15 (sCD15) in lung cancer tissue by using Leu-M1 antibody and MXKM-93 antibody, respectively, and then assessed the relationship between their expression and the patient outcome. Lung cancer tissue expression of CD15 was significantly higher in adenocarcinoma (55.9%) and squamous cell carcinoma (44.7%) than in small cell carcinoma (10%) (p=0.01, p=0.006). Expression of sCD15 was significantly higher in adenocarcinoma (52.9%) than in squamous cell carcinoma (10.5%) or small cell carcinoma (10%) (p<0. 0001, p=0.016). No association was found between CD15 expression and clinical stage, but sCD15 expression increased with clinical stage (stage I+II vs. III+IV: 16.7% vs. 39.6%; p=0.049). Expression of CD15 (1.5%) was significantly lower than expression of sCD15 (12.3%) in normal surrounding tissue. Examination of associations with outcome in NSCLC revealed that expression of sCD15 in resected cases, and expression of CD15 in non-resected cases were significantly correlated with shortening of median survival time (p<0.05). When associations with prognostic factors were assessed by univariate analysis, expression of sCD15 was found to be correlated with distant metastasis, and expression of CD15 with decrease in performance status (PS). In the multivariate analysis by the Cox proportional hazard model, sCD15 and CD15 negativity contributed to longer survival time after PS and clinical stage. The results of a combination assay of CD15 and sCD15 showed that expression of both carbohydrate antigens significantly shortened survival time in both the resected and non-resected group (log-rank test, p<0.05). This combination assay also appeared to be extremely useful in predicting the outcome in all clinical stages of NSCLC.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Animals; Antigens, Tumor-Associated, Carbohydrate; Carcinoma, Non-Small-Cell Lung; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Disease Progression; Female; Humans; Immunohistochemistry; Lewis X Antigen; Lung Neoplasms; Male; Middle Aged; Neoplasm Staging; Oligosaccharides; Prognosis; Rabbits; Risk Factors; Sialyl Lewis X Antigen; Survival Analysis

Alpha(1,3)- and alpha(1,4)-fucosylated oligosaccharides such as sialyl-Lewis(x) (sialyl-Le(x)) and sialyl-Lewis(a) (sialyl-Le(a)) have been reported to participate in tumor cell adhesion to activated endothelium. We examined by cytofluorometry the expression of Le(x), sialyl-Le(x), sialyl-Le(x) dimeric, Le(a), and sialyl-Le(a) on the surface of two human lung adenocarcinoma cell lines with different lung colonization potential. High expression levels of all of these antigens were detected in the metastatic HAL-8Luc cells, whereas the closely related nonmetastatic HAL-24Luc cells only expressed the sialyl-Le(a) and sialyl-Le(x) dimeric antigens, both at lower level than in HAL-8Luc cells. Five alpha(1,3)-fucosyltransferases (alpha(1,3)-Fuc-T) controlling the synthesis of these molecules have been identified to date (Fuc-TIII-Fuc-IVII). The expression of these five genes was also higher in the metastatic cells than in the nonmetastatic counterparts as was shown by Northern blot analysis. In vitro adhesion assays showed that only the metastatic cell line adheres significantly to E-selectin-expressing human endothelial cells. Moreover, monoclonal antibody (mAb) blockade of E-selectin completely abolished tumor cell binding. Adhesion inhibition experiments using mAbs against sialylated fucosylated oligosaccharides expressed on tumor cells indicated that these antigens are involved in the binding. Anti-sialyl-Lex(x) mAb (CSLEX-1) inhibited adhesion by 85%; it had the most pronounced inhibitory effect. These findings suggest that the overexpression of alpha(1,3)-Fuc-T genes in the metastatic HAL-8Luc cells, compared with HAL-24Luc cells, results in an enhanced surface display of fucosylated oligosaccharides, which contributes to the adhesive capacity of these cells to the activated endothelium and correlates with their lung colonization potential.

Topics: Adenocarcinoma; Animals; Antigens, Neoplasm; Antigens, Surface; Blotting, Northern; CA-19-9 Antigen; Cell Adhesion; E-Selectin; Endothelium; Flow Cytometry; Fucosyltransferases; Gangliosides; Gene Expression; Humans; Lewis X Antigen; Lung Neoplasms; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Metastasis; Oligosaccharides; Sialyl Lewis X Antigen; Tumor Cells, Cultured

The most common clinical form of lung cancer is a disseminated disease with distant metastases; several years of cancer progression precede presentation, and this ultimately limits the efficacy of curative therapy. In this immunohistochemical study, we examined a mucinous adenocarcinoma cell line, maintained by xenogeneic transplantation, and a spontaneous metastatic variant which produces distant tumors (in liver, spleen and kidney). The aim was to investigate possible parameters which characterize the metastatic process. Histopathological comparison between the two subcutaneous transplanted tumor lines showed that both lines presented a similar cellular morphology, a different pattern of cellular growth and an increased vascularization in the metastatic line with respect to its parent. All the tumor sections expressed differential immune reactivity with monoclonal antibodies against Lewis y (MAb C14), sialyl-Lewis x (MAb SNH3) and Lewis x (MAb FH2) determinants. Neither expressed MUC 1 mucins detectable with monoclonal antibodies reactive with the mucin protein core (MAbs C595 and SM3) nor was carcinoembryonic antigen (MAb C365) expressed. Neoplastic cells were reactive with an anti-pan cytokeratin monoclonal antibody confirming their epithelial histogenesis. Our findings have been evaluated with respect to defining metastatic phenotypes in lung cancer by examination of distinct histopathological and immunological parameters.

Topics: Adenocarcinoma, Mucinous; Animals; Antibodies, Monoclonal; Apoptosis; Biomarkers, Tumor; Carcinoembryonic Antigen; Cell Nucleus; Cytoplasm; Gangliosides; Humans; Image Processing, Computer-Assisted; Immunoenzyme Techniques; Injections, Subcutaneous; Kidney Neoplasms; Lewis Blood Group Antigens; Lewis X Antigen; Liver Neoplasms, Experimental; Lung Neoplasms; Mice; Mice, Nude; Mucin-1; Mucins; Neoplasm Proteins; Neoplasm Transplantation; Neoplastic Stem Cells; Oligopeptides; Peptide Fragments; Phenotype; Sialyl Lewis X Antigen; Splenic Neoplasms; Transplantation, Heterologous; Tumor Cells, Cultured

To elucidate the early events of blood-borne metastasis under actual blood flow, real-time trafficking of RAW117 large cell lymphoma cells, namely parental RAW117-P and liver-metastatic RAW117-H10 cells, was investigated using positron emission tomography (PET). Both types of cells accumulated in the liver immediately after injection via the portal vein, and were eliminated from the liver time-dependently. The elimination rate of RAW117-H10 cells, however, was slower than that of RAW117-P cells, suggesting that RAW117-H10 cells interact more strongly with hepatic sinusoidal endothelium than the parental cells. This result correlated with the metastatic potential of these cells: RAW117-H10 cells metastasized in the liver to a greater extent than RAW117-P cells after injection via this route. To investigate the role of sialylglycoconjugates in the interaction of RAW117-H10 cells with the hepatic endothelium after injection via the portal vein, the trafficking of RAW117-H10 cells was examined after the cells had been treated with sialidase. The elimination rate of RAW117-H10 cells from liver was observed to be greatly accelerated by sialidase treatment. To elucidate what kind of sialylglycoconjugates is related to this phenomenon, we analyzed the distribution of sialyl Lewis A and sialyl Lewis X antigens of both sublines of RAW117 by using flow cytometry. RAW117-H10 cells were found to express a much higher level of sialyl Lewis A than RAW117-P cells, whereas the amount of sialyl Lewis X did not differ significantly. These findings suggest that some sialylglycoconjugates, perhaps sialyl Lewis A in particular, play an important role in the initial interaction of RAW117-H10 cells with the hepatic endothelium, leading to metastasis.

Topics: Animals; Antigens, Neoplasm; CA-19-9 Antigen; Cell Adhesion; Cell Movement; Female; Flow Cytometry; G(M1) Ganglioside; Gangliosides; Injections, Intravenous; Liver Neoplasms; Lung Neoplasms; Lymphoma, Large B-Cell, Diffuse; Mice; Mice, Inbred BALB C; Neoplasm Metastasis; Neoplasm Transplantation; Neoplastic Cells, Circulating; Neuraminidase; Portal Vein; Sialyl Lewis X Antigen; Tomography, Emission-Computed; Tumor Cells, Cultured

Biosynthesis of sialyl Lewis(x) (sLe(x)) requires a sialyltransferase for alpha-2,3-sialylation and a fucosyltransferase for alpha-1,3-fucosylation. To date, five human alpha-1,3-fucosyltransferase (Fuc-T) genes and five human alpha-2,3-sialyltransferase (ST) genes have been cloned. However, it is not known which enzyme is mainly responsible for sLe(x) synthesis.. Three hundred thirteen patients with nonsmall cell lung carcinoma who had a curative tumor resection were the subjects of this study. Using tumor tissues fixed in formaldehyde, amplification of genomic DNA of Fuc-T and ST was performed by PCR and correlated with sLe(x) staining and patient prognosis.. The frequency of strong ST3N and Fuc-TVII amplification was significantly higher than that of STZ, ST4, Fuc-TIII, Fuc-TV, and Fuc-TVI amplification (P < 0.01). The frequency of sLe(x) staining was similar to ST3N and Fuc-TVII amplification. Survival of the patients whose tumors had strong amplification of both ST3N and Fuc-TVII was significantly shorter than that of patients whose tumors had no amplification of either gene (P < 0.01). In a multivariate analysis of survival, Fuc-TVII remained a statistically significant prognostic factor.. In lung carcinoma, ST3N and Fuc-TVII may both be related to sLe(x) synthesis, and Fuc-TVII is a more important indicator of poor prognosis.

Topics: Adult; Aged; Aged, 80 and over; beta-Galactoside alpha-2,3-Sialyltransferase; Carcinoma, Non-Small-Cell Lung; Female; Fucosyltransferases; Gene Amplification; Humans; Isoenzymes; Lung Neoplasms; Male; Middle Aged; Multivariate Analysis; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen; Sialyltransferases; Staining and Labeling; Survival Analysis

The levels of sialyl Lewis X-i antigen (SLX), which is one of the cancer-associated carbohydrate antigens, were evaluated in 83 malignant and 46 benign pleural effusions. SLX levels in pleural effusion due to lung adenocarcinoma were significantly higher than those due to benign diseases (p < 0.0001), lung cancer other than adenocarcinoma (p = 0.0052), and adenocarcinoma originating from other organs (p = 0.0492). According to receiver operating characteristic (ROC) curve analysis, the optimal cut-off level in the discrimination between malignant and benign pleural effusions was 92 U/ml, which gave a sensitivity of 57.1% and a specificity of 77.8%. The cut-off level of pleural effusion in patients with carcinomatous pleuritis might be higher than that of serum (38 U/ml).

Topics: Adenocarcinoma; Biomarkers, Tumor; Biopsy; Carcinoma, Small Cell; Carcinoma, Squamous Cell; Female; Humans; Lung Neoplasms; Male; Neoplasms; Oligosaccharides; Pleural Effusion, Malignant; Pleurisy; Radioimmunoassay; Reproducibility of Results; ROC Curve; Sensitivity and Specificity; Sialyl Lewis X Antigen

To evaluate the correlation between serum levels of sialyl Lewis X-i antigen and distant metastasis and survival in patients with non-small cell lung cancer (NSCLC), we measured the serum levels of the tumor marker in 371 patients with untreated NSCLC. The sialyl Lewis X-i antigen level was measured using a RIA kit. In patients with adenocarcinoma or other NSCLC subtypes, there was a correlation between serum sialyl Lewis X-i antigen and stage of the disease (P = 0.0001 and P = 0.0015, respectively). Levels of the marker varied significantly depending on the number of metastatic organs in adenocarcinoma (P = 0.0089) and in other NSCLC subtypes (P = 0.002). Univariate analysis showed that survival of NSCLC patients with high (more than 100 units/ml) sialyl Lewis X-i antigen levels was significantly poorer than that of patients with low antigen levels (P = 0.0001). Multivariate analysis using Cox's proportional hazard model showed that high sialyl Lewis X-i antigen levels correlated significantly with poor survival (P = 0.004). Our data suggest that a high serum level of sialyl Lewis X-i antigen seems to be an indicator of the presence of metastasis and might indicate the need for a careful investigation of all putative metastatic sites. The serum levels of sialyl Lewis X-i antigen may reflect the extension of metastasis and would be helpful in considering treatment options.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Non-Small-Cell Lung; Female; Humans; Lewis Blood Group Antigens; Lung Neoplasms; Male; Middle Aged; Neoplasm Metastasis; Neoplasm Staging; Oligosaccharides; Sialyl Lewis X Antigen; Survival Analysis; Time Factors

To date, five alpha-1,3-fucosyltransferase genes (Fuc-TIII, IV, V, VI, and VII) have been cloned. To examine the role of alpha-1,3-fucosyltransferase in the synthesis of sialyl Lewis x and the prognosis of lung cancer, PCR amplification of five fucosyltransferase genes and immunohistochemical staining of sialyl Lewis x were performed in 333 patients with lung cancer who underwent surgical resection from 1980 to 1993. The frequencies of Fuc-TIII/V, Fuc-TVI, Fuc-TIV, and Fuc-TVII expression were 9%, 26%, 75%, and 66%, respectively. The frequency of sialyl Lewis x expression (75%) was comparable to Fuc-TIV and Fuc-TVII expression. However, the grading of sialyl Lewis x staining correlated only with the grading of Fuc-TVII gene amplification. Survival of the patients whose tumors showed strong expression of Fuc-TIV and/or FucT-VII was significantly shorter than that of the patients whose tumors did not express either Fuc-TIV or Fuc-TVII. These results suggest that Fuc-TIV and Fuc-TVII expression may be of prognostic value among patients with lung cancer by participating in the biosynthesis of sialyl Lewis x.

Topics: Adult; Aged; Aged, 80 and over; Base Sequence; DNA, Neoplasm; Female; Fucosyltransferases; Gene Expression; Humans; Immunohistochemistry; Lung Neoplasms; Male; Middle Aged; Molecular Sequence Data; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen

The selectin class of adhesion molecules plays a critical role in facilitating leukocyte adhesion to and subsequent transmigration of endothelium. On this basis, selectins have been suggested to promote tumor cell attachment to endothelium, thereby facilitating metastasis of certain types of tumors, although direct evidence for such a role is lacking. To explore this hypothesis, two sets of transgenic mice were developed: TgnES, which constitutively expresses cell surface E-selectin in all tissues, under the control of the beta-actin promoter; and TgnEsol, which expresses truncated, soluble E-selectin in the liver, under the control of the alpha 1 antitrypsin promoter. B16F10 melanoma cells were stably transfected with alpha(1,3/1,4) fucosyltransferase-specific cDNA (B16F10ft), allowing them to express E-selectin ligands or with hygromycin resistance selection vector only B16F10hygro). Normal mice injected with B16F10ft and B16F10hygro and transgenic mice injected with B16F10hygro developed lung tumors exclusively. In contrast, TgnES mice injected with B16F10ft cells developed massive infiltrating liver tumors. B16F10ft cells injected into TgnEsol mice also formed liver tumors, but these grew more slowly, with a well-delineated, noninfiltrating distinct histologic pattern. These observations provide direct evidence that expression of E-selectin can redirect metastasis of tumor cells expressing appropriate ligands in vivo.

Topics: Animals; Base Sequence; CA-19-9 Antigen; E-Selectin; Fucosyltransferases; Gangliosides; Liver Neoplasms; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Transgenic; Molecular Sequence Data; Neoplasm Metastasis; Oligosaccharides; Peptide Fragments; Recombinant Fusion Proteins; Sialyl Lewis X Antigen

Our study demonstrates that synthetic sialyl Lewis X (SLex) as a ligand for selectins and fibronectin-derived RGDS peptide analogue [Ar(DRGDS)3] inhibits lung metastases produced by i.v. co-injection of B16-BL6 melanoma cells. To investigate the inhibitory mechanisms in a living animal, we performed positron-emission tomography (PET) analysis after i.v. injection of [2-18F]2-fluoro-2v-deoxy-D-glucose-labeled tumor cells with or without liposomal SLex or Ar(DRGDS)3. The real-time PET measurement for the first 120 min, started immediately after injection, showed that tumor-cell arrest, i.e., accumulation in the target organ (lung) was remarkably inhibited by liposomal SLex, but not inhibited by Ar(DRGDS)3 or liposomal Me-SLex, which is not recognized by selectins. In contrast, Ar(DRGDS)3 inhibited the invasion of B16-BL6 cells into reconstituted basement membrane (Matrigel) following tumor arrest, whereas SLex- or Me-SLex-entrapped liposomes did not affect tumor invasion. In the metastatic processes containing tumor-cell lodgement and arrest in the target organ followed by extravasation (invasion), SLex resulted in the inhibition of initial arrest of tumor cells, presumably tumor-endothelium interaction, while Ar(DRGDS)3 achieved inhibition of tumor invasion into basement membrane at later steps of the cascade, consequently leading to inhibition of metastasis. Thus, tumor-cell arrest in lungs in the metastatic processes must be precisely and properly controlled by different adhesion molecules at different stages, which are similar to those observed in leukocyte-endothelium interaction.

Topics: Amino Acid Sequence; Animals; Basement Membrane; Carbohydrate Sequence; Cell Adhesion; Female; Fibronectins; Integrins; Liposomes; Lung Neoplasms; Melanoma, Experimental; Mice; Mice, Inbred C57BL; Molecular Sequence Data; Neoplasm Invasiveness; Neoplastic Cells, Circulating; Oligopeptides; Oligosaccharides; Selectins; Sialyl Lewis X Antigen; Tomography, Emission-Computed

In order to improve the diagnosis of lung carcinoma, in which a complicated histologic pattern is present, the immunohistochemistry of 119 adenocarcinomas, 65 squamous cell carcinomas, 12 small cell carcinomas, 18 large cell carcinomas, and 15 metastatic adenocarcinoma in the lung were evaluated using a monoclonal antibody, KM195, against lung carcinoma, and compared with the immunohistochemical results using anti-human cytokeratin (CAM 5.2) and other monoclonal antibodies. Formalin-fixed, paraffin-embedded tissues were stained using the labeled streptavidin-biotin method. Extracts from fresh tissue homogenate, after fractionation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, were transferred by Western blotting and stained with KM195. The anti-lung adenocarcinoma, murine, monoclonal antibody KM195 (IgG), was positive in 107 of 119 adenocarcinomas (90%), in 15 of 18 large cell carcinoma (83%), in three of 65 squamous cell carcinomas (5%), 13 of 15 (87%) metastatic adenocarcinoma in the lung, and was negative in 12 small cell carcinomas (P < 0.001). KM195-bound protein of primary and metastatic adenocarcinoma in the lung cases concentrated at about 40 kDa. In contrast, CAM 5.2 was positive in 52 of 67 (78%) adenocarcinomas, 10 of 62 (16%) squamous cell carcinomas, and was negative in six small cell carcinomas. These results suggest that the immunohistochemistry for KM195 may be a more useful marker over CAM 5.2 for the diagnosis of pulmonary adenocarcinoma.

Topics: Adenocarcinoma; Antibodies, Monoclonal; Bacterial Proteins; Biomarkers, Tumor; Biotin; Blotting, Western; Carcinoembryonic Antigen; Humans; Immunohistochemistry; Keratins; Lung; Lung Neoplasms; Oligosaccharides; Paraffin Embedding; Reference Values; Sialyl Lewis X Antigen; Tissue Fixation