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

Esophageal (OC), gastric (GC) and colorectal (CRC) cancers are amongst the digestive track tumors with higher incidence and mortality due to significant molecular heterogeneity. This constitutes a major challenge for patients' management at different levels, including non-invasive detection of the disease, prognostication, therapy selection, patient's follow-up and the introduction of improved and safer therapeutics. Nevertheless, important milestones have been accomplished pursuing the goal of molecular-based precision oncology. Over the past five years, high-throughput technologies have been used to interrogate tumors of distinct clinicopathological natures, generating large-scale biological datasets (

Topics: Antigens, Tumor-Associated, Carbohydrate; Colorectal Neoplasms; Esophageal Neoplasms; Glycomics; Glycoproteins; Glycosylation; Humans; Precision Medicine; Proteomics; Sialyl Lewis X Antigen; Stomach Neoplasms

Aberrant expression of glycoconjugates occurs during malignant transformation of cancer cells. Overexpression of sialoglycoconjugates in particular may play an important role in the progression, i.e., invasion or metastasis, of cancer. Various types of sialoglycoconjugates have been investigated to clarify their biological significance and clinical utility in diagnosing and treating colorectal cancer. This review focuses specifically on expression of mucin (MUC) 1 and it suggests that MUC1 with the specific structure of a sialo-oligosaccharide has biological significance in determining the metastatic potential of colorectal cancer cells and clinicopathological utility in evaluating the effectiveness of treatments and the prognosis for patients with colorectal cancer. Further studies are expected to contribute to the expanded use of cancer-associated sialoglycoconjugates in cancer diagnosis and therapy.

Topics: Antigens, Tumor-Associated, Carbohydrate; CA-19-9 Antigen; Colorectal Neoplasms; Diagnostic Imaging; Gene Expression Regulation, Neoplastic; Glycoconjugates; Humans; Mucin-1; Neoplasm Invasiveness; Neoplasm Metastasis; Oligosaccharides; Prognosis; Sialic Acids; Sialyl Lewis X Antigen

he colonization of the liver by colorectal cancer (CRC) cells is a complicated process which includes many stages, until macrometastases occur. The entrapment of malignant cells within the hepatic sinusoids and their interactions with resident non-parenchymal cells are considered very important for the whole metastatic sequence. In the sinusoids, cell connection and signalling is mediated by multiple cell adhesion molecules, such as the selectins. The three members of the selectin family, E-, P- and L-selectin, in conjunction with sialylated Lewis ligands and CD44 variants, regulate colorectal cell communication and adhesion with platelets, leucocytes, sinusoidal endothelial cells and stellate cells. Their role in CRC liver metastases has been investigated in animal models and human tissue, in vivo and in vitro, in static and shear flow conditions, and their key-function in several molecular pathways has been displayed. Therefore, trials have already commenced aiming to exploit selectins and their ligands in the treatment of benign and malignant diseases. Multiple pharmacological agents have been developed that are being tested for potential therapeutic applications.

Topics: Carbohydrate Conformation; Carbohydrate Sequence; Colorectal Neoplasms; Hexanes; Humans; Lewis X Antigen; Ligands; Liver Neoplasms; Mannose; Molecular Sequence Data; Molecular Structure; Protein Isoforms; Selectins; Sialyl Lewis X Antigen

Topics: Carcinoma; Cell Adhesion Molecules; Cell Movement; Colorectal Neoplasms; Glycosylation; Humans; Integrins; Kalinin; Liver Neoplasms; Neoplasm Invasiveness; Neoplasms; Oligosaccharides; Sialyl Lewis X Antigen

CEA and CA19-9 are the two most common tumor markers for colorectal cancer that are currently utilized clinically. The positive rate of CEA is 40-60% and that of CA19-9 is 30-50%. Simultaneous use of the two markers is useful in evaluating the therapeutic effect and monitoring the recurrence of advanced colorectal cancer. Surgical specimens may also provide useful information for the appropriate treatment of patients. Using surgically resected lymph nodes, we examined micrometastasis to assess the spread of the cancer cells and the malignant potential of colorectal cancer. Immunohistochemical analysis using anti-cytokeratin antibody revealed no significant impact of micrometastasis on patient prognosis, while RT-PCR assay using CEA as a genetic marker suggested a positive value in predicting a rapid recurrence. Among various molecular markers, we found that CDC25B phosphatase was a powerful prognostic factor for colorectal cancer. Diagnosis of the existence and malignant potential of cancer cells, together with serum tumor marker levels, may help to construct a more useful system for the better treatment of colorectal cancer.

Topics: Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; cdc25 Phosphatases; Cell Adhesion Molecules; Cell Cycle Proteins; Colorectal Neoplasms; DCC Receptor; Female; Genes, src; Humans; Keratin-19; Keratins; Male; Oligosaccharides; Receptors, Cell Surface; Sialyl Lewis X Antigen; Tumor Suppressor Proteins

Cimetidine has been shown to have beneficial effects in colorectal cancer patients. In this study, a total of 64 colorectal cancer patients who received curative operation were examined for the effects of cimetidine treatment on survival and recurrence. The cimetidine group was given 800 mg day(-1) of cimetidine orally together with 200 mg day(-1) of 5-fluorouracil, while the control group received 5-fluorouracil alone. The treatment was initiated 2 weeks after the operation and terminated after 1 year. Robust beneficial effects of cimetidine were noted: the 10-year survival rate of the cimetidine group was 84.6% whereas that of control group was 49.8% (P<0.0001). According to our previous observations that cimetidine blocked the expression of E-selectin on vascular endothelium and inhibited the adhesion of cancer cells to the endothelium, we have further stratified the patients according to the expression levels of sialyl Lewis antigens X (sL(x)) and A (sL(a)). We found that cimetidine treatment was particularly effective in patients whose tumour had higher sL(x) and sL(a) antigen levels. For example, the 10-year cumulative survival rate of the cimetidine group with higher CSLEX staining, recognizing sL(x), of tumours was 95.5%, whereas that of control group was 35.1% (P=0.0001). In contrast, in the group of patients with no or low levels CSLEX staining, cimetidine did not show significant beneficial effect (the 10-year survival rate of the cimetidine group was 70.0% and that of control group was 85.7% (P=n.s.)). These results clearly indicate that cimetidine treatment dramatically improved survival in colorectal cancer patients with tumour cells expressing high levels of sL(x) and sL(a).

Topics: Administration, Oral; Adult; Aged; Antimetabolites, Antineoplastic; Biomarkers, Tumor; CA-19-9 Antigen; Cell Adhesion; Chemotherapy, Adjuvant; Cimetidine; Colorectal Neoplasms; Female; Fluorouracil; Gangliosides; Histamine H2 Antagonists; Humans; Male; Middle Aged; Neoplasm Metastasis; Neoplasm Recurrence, Local; Oligosaccharides; Sialyl Lewis X Antigen; Survival Analysis; Treatment Outcome

Circulating tumor cells (CTCs) have been detected by us and others in cancer patient blood. However, little is known about how to specifically capture and deactivate CTCs in vivo, which may lead to successful metastasis prevention in asymptomatic cancer survivors after surgery. We hypothesize that the dual antibody conjugates may have the advantage of capturing CTCs specifically over their single antibody counterparts. Here we show that the surface-functionalized dendrimers can be sequentially coated with two antibodies directed to surface biomarkers (EpCAM and Slex) of human colorectal CTCs. The dual antibody-coated dendrimers exhibit a significantly enhanced specificity in capturing CTCs in the presence of interfering blood cells, and in both eight-patient bloods and nude mice administered with the labeled CTCs in comparison to their single antibody-coated counterparts. The dual antibody-coated conjugates down-regulate the captured CTCs. This study provides the first conceptual evidence that two antibodies can be biocompatibly conjugated to a nanomaterial to capture and down-regulate CTCs in vivo with the enhanced specificity.

Topics: Animals; Antibodies; Antigens, Neoplasm; Cell Adhesion Molecules; Colorectal Neoplasms; Dendrimers; Epithelial Cell Adhesion Molecule; HL-60 Cells; HT29 Cells; Humans; Mice, Nude; Nanostructures; Neoplastic Cells, Circulating; Oligosaccharides; Sialyl Lewis X Antigen

Treatment decisions in colorectal cancer subsequent to surgery are based mainly on the TNM system. There is a need to establish novel prognostic markers based on the molecular characterization of tumor cells. Evidence exists that sialyl Le(X) expression is correlated with an unfavorable outcome in colorectal cancer. The aim of this study was to establish a simple sialyl Le(X) staining score and to determine a potential correlation with the prognosis in a series of advanced colorectal carcinoma patients.. In order to implement routine use of sialyl Le(X) immunohistology, we established a new, easily reproducible score and defined a cutoff which discriminated groups with better or worse outcome, respectively. We then correlated sialyl Le(X) expression of 215 UICC stage III and IV patients with disease-free and cancer-related survival.. A five-stage score could be established based on automated immunohistochemical stainings. Using a statistical model, we calculated a cutoff to discriminate between weak and strong staining positivity of sialyl Le(X). Patients with strong positive specimens had a worse cancer-related survival (P = 0.004) but no difference was observed for disease-free survival (P = 0.352).. These results demonstrate a strong correlation between high sialyl Le(X)-expression in colorectal carcinomas and cancer-related survival. Our highly standardized and easy-to-use staining score is suitable for routine use and hence it could be recommended to evaluate sialyl Le(X)-expression as part of the standard histopathological analysis of colorectal carcinomas and to validate the score prospectively based on a larger population.

Topics: Aged; Biomarkers, Tumor; Colorectal Neoplasms; Female; Follow-Up Studies; Humans; Immunoenzyme Techniques; Lymph Nodes; Lymphatic Metastasis; Male; Neoplasm Grading; Neoplasm Staging; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen; Survival Rate

Sialyl Lewis x (sLex) is a selectin ligand whose overexpression in epithelial cancers mediates metastasis formation. The molecular basis of sLex biosynthesis in colon cancer tissues is still unclear. The prerequisite for therapeutic approaches aimed at sLex down-regulation in cancer, is the identification of rate-limiting steps in its biosynthesis. We have studied the role of α1,3-fucosyltransferases (Fuc-Ts) potentially involved in sLex biosynthesis in specimens of normal and cancer colon as well as in experimental systems. We found that: (i) in colon cancer, but not in normal mucosa where the antigen was poorly expressed, sLex correlated with a Fuc-T which, like Fuc-TVI, was active on 3'sialyllactosamine at a low concentration (Fuc-T(SLN)); (ii) competitive RT-PCR analysis revealed that the level of Fuc-T mRNA expression in both normal and cancer colon was Fuc-TVI>Fuc-TIII>Fuc-TIV; Fuc-TV and Fuc-TVII expression was negligible; (iii) sLex was expressed only by the gastrointestinal cell lines displaying both Fuc-TVI mRNA and Fuc-T(SLN) activity, but not by those expressing only Fuc-TIII mRNA; (iv) transfection with Fuc-TVI cDNA, but not with Fuc-TIII cDNA, induced sLex expression in gastrointestinal cell lines; (v) Fuc-TVI knock-down with specific siRNA induced down-regulation of Fuc-TVI mRNA and Fuc-T(SLN) activity and a dramatic inhibition of sLex expression. These data indicate that in colon cancer tissues Fuc-TVI is a key regulator of sLex biosynthesis which can be the target of RNA-interference-based gene knock-down approaches.

Topics: Base Sequence; Colorectal Neoplasms; DNA, Complementary; Fucosyltransferases; Gene Expression Regulation, Neoplastic; Gene Knockdown Techniques; Glycosylation; Humans; Molecular Sequence Data; Neoplasm Metastasis; Oligosaccharides; RNA Interference; Selectins; Sialyl Lewis X Antigen; Transfection; Tumor Cells, Cultured

The glycoconjugates expressed by cancer cells frequently contain sialylated oligosaccharide chains. Among these oligosaccharides the sialyl Lewis a (sLe(a)) and sialyl Lewis x (sLe(x)) antigens are found to be overexpressed in tumours of different origin. The current study assesses sLe(a) and sLe(x) expression in different colorectal specimens in order to establish the correlation of these biomarkers with both malignant transformation of colorectal mucosa and the progression of colorectal cancer (CRC). Healthy disease-free and inflammatory mucosa specimens showed no presence of the antigens. sLe(a) was expressed in 6.7% of the healthy tissue from CRC patients, in 20.8% of the adenomas, and in 33.3% and 42.6% of the transitional tissue and tumour tissue, respectively. sLe(x) expression was observed in 6.7% of the healthy tissue from CRC patients, in 27.0% of the adenomas, and in 25.6% and 74.8% of the transitional and the tumour tissue, respectively. The expression of the sLe(a) and sLe(x) antigens was correlated in adenomas, as well as in healthy and tumour tissue from CRC. Moreover, the high expression of sLe(x) in adenomas was correlated with a high degree of dysplasia (p=0.042). Finally, the survival analysis suggested that sLe(a) expression may be a prognostic factor for predicting disease-free survival in colorectal cancer (p=0.012).

Topics: Adenocarcinoma; Adenoma; Biomarkers, Tumor; CA-19-9 Antigen; Cell Transformation, Neoplastic; Colorectal Neoplasms; Disease-Free Survival; Humans; Immunohistochemistry; Kaplan-Meier Estimate; Neoplasm Staging; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen

Changes in the expression levels of alpha1,3 FucT-VII and carbohydrate product SLe X have been reported in certain types of malignant transformations. However, the specific role of FucT-VII in human colorectal carcinoma is still not clear. We evaluated the expression of FucT-VII in tumor specimens from 30 colorectal carcinoma patients by RT-PCR and immunohistochemistry. The alteration of metastatic potential of LOVO cells before and after alpha1,3 FucT-VII gene transfection was also assayed. The expression change of glycoprotein CD24 and carbohydrate antigen SLe X after stable transfection of LOVO was also examined in vitro. Higher mRNA and protein expression of FucT-VII were observed in colorectal tumors compared with adjacent normal ones. The cell chemotactic migration and invasion were enhanced after alpha1,3 FucT-VII transfection in LOVO cells. There was a positive correlation between alpha1,3 FucT-VII mRNA expression and lymph node status (p=0.009) and AJCC stage (p=0.007), similar correlation was also observed at protein level (p=0.042 and 0.022, respectively). Overexpression of alpha1,3 FucT-VII promoted the expression of CD24 and SLe X in LOVO cells. Our findings suggest that alpha1,3 FucT-VII might play a role in colorectal carcinoma metastases by promoting the carbohydration of glycoprotein CD24.

Topics: Apoptosis; Blotting, Western; CD24 Antigen; Cell Adhesion; Cell Movement; Cell Proliferation; Colon; Colorectal Neoplasms; Flow Cytometry; Fucosyltransferases; Humans; Immunoenzyme Techniques; Lymphatic Metastasis; Middle Aged; Oligosaccharides; Rectum; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sialyl Lewis X Antigen

To evaluate the efficacy and safety of a regimen using Irinotecan, 5FU and Leucovorin for patients with advanced or recurrent colorectal cancer.. Irinotecan (75 mg/m(2)) was administered biweekly, while 5FU (600 mg/m(2)) and Leucovorin (250 mg/m(2)) were administered weekly, for 6 weeks.. The 21 consecutive patients subjected to this regimen showed a good response rate (43%) with minimal toxicity (incidence of grade 3/4: leukopenia and neutropenia, 5%, respectively, and vomiting, 10%). The mean survival time of all 21 patients was 15.7 months. This regimen could be a valid option for patients with advanced colorectal cancer, especially those seeking a good QoL (quality of life) for the remainder of their lives. We evaluated the expression of thymidylate synthase (TS), dihydropyrimidine dehydrogenase (DPD), thymidine phosphorylase (TP) and orotate phosphoribosyl transferase (OPRT) mRNAs, and sialyl Lewis X on formalin-fixed, paraffin-embedded colorectal tumor samples. Expression of TS mRNA or sialyl Lewis X was negatively correlated with the response from chemotherapy. Patients with low DPD mRNA expression in the tumor showed a significant longer survival than those with high expression. In patients with high TP mRNA expression, there was a tendency towards a high incidence of leukopenia.. Some predictive factors elucidated in this study could contribute to the progress of the tumor-biology based, individualized chemotherapy for colorectal cancer patients.

Topics: Adenocarcinoma; Adult; Aged; Antigens, Tumor-Associated, Carbohydrate; Antineoplastic Combined Chemotherapy Protocols; Biomarkers, Tumor; Camptothecin; Colorectal Neoplasms; Dihydrouracil Dehydrogenase (NADP); Female; Fluorouracil; Humans; Irinotecan; Leucovorin; Lewis X Antigen; Male; Middle Aged; Oligosaccharides; Orotate Phosphoribosyltransferase; Prognosis; Sialyl Lewis X Antigen; Survival Rate; Thymidine Phosphorylase; Thymidylate Synthase; Vitamin B Complex

1- to detect alpha 1-acid glycoprotein (AGP) and sialyl Lewis x (sLex) in colorectal malignant, benign and normal samples; 2- to isolate AGP from colorectal cancer and 3- to study its immunoreactivity with an anti-sLex monoclonal antibody (MAb).. tissue and serum samples from 88 patients with colorectal cancer, 22 adenomas and 23 normal were included. Expression of AGP and sLex was studied by immunohistochemistry (IHC); isolation approach: AGP was precipitated with ammonium sulphate and immunoprecipitated with anti-AGP MAb. The immune complex formed was isolated by protein A-Sepharose CL-4B affinity chromatography and further eluted; fractions were analysed by SDS-PAGE and Western-blot. Statistical analysis was performed by means of Principal Component Analysis.. By Western blot employing anti-AGP MAb and sLex MAbs, isolated fractions from malignant samples showed a band at about 45 kD. IHC revealed that AGP was expressed in 70% of colorectal carcinoma samples, 50% of benign and 35% of normals. SLex was detected in 31% of malignant samples, 41% of benign and in one normal sample. In malignant samples, AGP reaction comprised the whole specimen with a strong and homogeneous staining while normal and benign samples showed a restricted reaction. In cancer, sLex expression consisted in an intense reactivity in membrane, cellular debris and some cytoplasmic foci while normal and benign samples were occasionally stained. A statistically significant positive correlation was found between AGP and sLex expression. Serum AGP levels were measured by radial immunodiffusion and statistical comparative analysis with tissue expression did not show a correlation between both parameters.. AGP may constitute a carrier of sLex in colorectal cancer.

Topics: Adenoma; Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Carcinoma; Colon; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Male; Middle Aged; Oligosaccharides; Orosomucoid; Sialyl Lewis X Antigen

Sialyl Lewis(x) (sLe(x)) is one ligand for E selectin (CD62E), a glycoprotein that is expressed on activated endothelial cells. Adhesion mediated by endothelial E selectin and sLe(x) expressed on human tumor cells could be relevant for the development of metastases. The aim of this study was to investigate whether or not a correlation exists between pre-operative levels of ganglioside serum sLe(x) and disease-free interval and survival in colorectal cancer patients.. Thirty Duke's B and 52 Duke's C patients undergoing resection for colorectal cancer were studied. The median follow-up time was 34.8 months. A pool of 57 sera from normal subjects was used as an Internal Normal Standard (INS). sLe(x) analyses were performed by a thin layer chromatography (TLC) immunostaining technique. Results were expressed as the ratio (R) between bands of INS and bands from each neoplastic serum.. The median R value was 0.80 in normal subjects, 0.70 in Duke's B patients and 1.0 in Duke's C patients. No significant differences were detected between sLe(x) concentrations in sera from normal and neoplastic subjects (p = 0.1). Using an arbitrary cutoff of R = 0.9, the mean disease-free interval in the whole series was 47.4 months for R <0.9 and 126.0 months for R > or = 0.9 (p = 0.04). The survival time was 76.8 months for patients with R values <0.9 and 156.3 months for patients with R values > or =0.9 (p = 0.1).. High serum levels of ganglioside sLe(x) significantly correlate with a favorable prognosis and with a lower occurrence of metastases. It is conceivable that soluble sLe(x) may compete with membrane-bound sLe(x) in the course of interactions between activated endothelium and tumor cells that have reached the circulation.

Topics: Aged; Biomarkers, Tumor; Case-Control Studies; Chromatography, Thin Layer; Colorectal Neoplasms; Disease-Free Survival; Female; Humans; Italy; Lewis X Antigen; Male; Neoplasm Metastasis; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen; Survival Analysis

Carbohydrate expression of cancer cells is closely related to the metastatic nature of colorectal cancer. In the present study we investigated the relevance of carbohydrate expression profiles of colorectal cancer cells in the primary lesion to metastatic distribution patterns as well as prognosis in 134 cases. Carbohydrate expression was estimated by histochemistry with 17 kinds of lectins and 3 kinds of Lewis-related monoclonal antibodies (MAbs), and correlations between the staining and clinicopathological parameters were examined. The results showed that lymphatic invasion, lymph node metastasis, and peritoneal metastasis correlated with staining with lectins that bind galactose/N-acetylgalactosamine residues (Gal/GalNAc) such as Maclura pomifera (MPA), Arachis hypogaea (PNA), Helix pomatia (HPA), and Vicia villosa (VVA). In contrast, hepatic metastasis correlated with staining with Anguilla anguilla lectin (AAA), anti-LewisX (LEX-2), anti-sialyl Lewisa (NS 19-9), and anti-sialyl-dimeric LewisX (FH-6) MAbs, all of which bind preferentially to fucosylated carbohydrate chains. The five-year survival rate of patients was related to the staining of cancers with MPA, HPA, FH-6 or NS19-9, and MPA- and FH-6 staining were independent prognostic factors. We conclude that carbohydrate expression profiles of cancer cells are relevant to the route of tumor cell dissemination, metastatic pattern as well as prognosis of colorectal cancer.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Animals; Antibodies, Monoclonal; Antigens, Neoplasm; Carbohydrate Metabolism; Carbohydrate Sequence; Carbohydrates; Colorectal Neoplasms; Female; Humans; Lectins; Lewis X Antigen; Liver Neoplasms; Lymphatic Metastasis; Male; Middle Aged; Molecular Sequence Data; Neoplasm Invasiveness; Neoplasm Metastasis; Oligosaccharides; Peritoneal Neoplasms; Prognosis; Prospective Studies; Sialyl Lewis X Antigen; Staining and Labeling

The significance of MUC1, MUC2 and sialylated Lewis blood group antigens as prognostic markers in colorectal adenocarcinoma was investigated in a large series of patients because previous investigations revealed inconsistent results due to unrelated tumour samples from different patient groups and methodological differences.. Tissues from 243 patients with colorectal adenocarcinoma were stained immunohistochemically. MUC1 showed a strong immunoreactivity (in more than 35% of the tumour area) in 32.5%, MUC2 in 51.0%, sialyl-Lewis(x) in 67.9% and sialyl-Lewis(a) in 73.7% of the cases, respectively. MUC1 immunoreactivity displayed a significant correlation with tumour progression as reflected by advancing pTNM staging and poor differentiation. MUC2 expression was significantly stronger in mucinous adenocarcinomas. Sialyl-Lewis(x) immunostaining correlated with the extent of lymph node metastasis as well as low cytological differentiation. According to univariate and multivariate analysis (P < 0.0001) only MUC1 reactivity represented a marker of worse survival probability, opposed to the sialylated Lewis antigens that did not exert a predictive value.. According to our data, MUC1 and sialyl-Lewis(x) immunoreactivity exhibit statistically significant correlations with established markers of tumour progression. However, only MUC1 presents as an independent prognostic factor of colorectal adenocarcinoma.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Antigens, Tumor-Associated, Carbohydrate; CA-19-9 Antigen; Colorectal Neoplasms; Female; Follow-Up Studies; Gangliosides; Humans; Immunohistochemistry; Male; Middle Aged; Mucin-1; Mucin-2; Mucins; Neoplasm Staging; Oligosaccharides; Predictive Value of Tests; Prognosis; Sialyl Lewis X Antigen; Survival Analysis; Survival Rate; Time Factors

This study aimed to determine whether sialyl Lewis(a) (Le(a)), sialyl Lewis(x )(Le(x)), or sialyl Tn antigen expression could identify a subset of node-negative colorectal cancer patients that are at high risk for recurrence after curative surgery. Tumor tissue samples from 90 patients with node-negative colorectal cancer, who had undergone surgical resection, were analyzed immunohistochemically for the expression of each antigen. Patients were classified as having low or high antigen expression depending on whether more or less than 40% of the field showed positive staining. The main outcome measure for each variable was disease-free interval. Sialyl Le(a), sialyl Le(x), and sialyl Tn antigens were expressed in 53 (58.9%), 41 (45.6%), and 34 (37.8%) carcinomas, respectively. The median follow-up was 83.5 months. Patients with high sialyl Le(x) expression had shorter disease-free intervals than those with low sialyl Le(x) expression (P = 0.0041); the expression of sialyl Le(a) or sialyl Tn antigens did not show a significant relationship with disease-free survival. Cox's regression analysis revealed that sialyl Le(x) expression was an independent predictor for disease-free survival, separate from T factor or tumor location. High sialyl Le(x) expression may be useful in identifying a subset of node-negative colorectal cancer patients who are at high risk for recurrence.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Colorectal Neoplasms; Disease-Free Survival; Female; Humans; Immunoenzyme Techniques; Lewis Blood Group Antigens; Lymph Nodes; Male; Middle Aged; Neoplasm Invasiveness; Neoplasm Staging; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen; Survival Rate

Preoperative serum levels of sialyl Lewis(a) (CA 19-9), sialyl Lewis(x) (SLX), and sialyl Tn (STN) antigens in colorectal cancer patients were examined to establish predictive factors for serum levels of these antigens compared with carcinoembryonic antigen (CEA). A total of 308 patients who underwent resection for a colorectal cancer were divided into low and high antigen groups (higher or lower than a selected diagnostic-based cutoff value). The cutoff values were 37 U/ml for CA19-9, 38 U/ml for SLX, 45 U/ml for STN, and 2.5 ng/ml for CEA. The American Joint Committee on Cancer Classification and Stage grouping was used to classify the tumors. Statistical tests were conducted using univariate and multivariate logistic regression analyses. For CA19-9, 81 patients (26.3%) were assigned to the high antigen group: for SLX, 39 (12.7%); for STN, 33 (10.7%); and for CEA, 133 (43.2%). Multivariate logistic regression analysis revealed that predictive factors associated with high antigen levels were female sex (odds ratio [OR], 1.78 vs male sex), T4 (OR, 3.26 vs T1/T2), and M1 (OR, 3.35 vs M0) for CA19-9; M1 (OR, 6.40 vs M0) for SLX; mucinous carcinoma (OR, 8.45 vs well differentiated adenocarcinoma) and M1 (OR, 8.24 vs M0) for STN; and mucinous carcinoma (OR, 7.21 vs well differentiated adenocarcinoma), T3/T4 (OR, 3.84/4.18, respectively, vs T1/T2), and M1 (OR, 6.39 vs M0) for CEA. In conclusion, high serum levels of CA19-9, SLX, and STN are strongly associated with distant metastasis. In addition, high serum levels of CA19-9 may be an independent predictor for female gender and T4, and high serum levels of STN may be an independent predictor for mucinous carcinoma.

Topics: Aged; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Colorectal Neoplasms; Female; Gangliosides; Humans; Logistic Models; Male; Middle Aged; Multivariate Analysis; Neoplasm Recurrence, Local; Oligosaccharides; Predictive Value of Tests; Sialyl Lewis X Antigen

Two types of colorectal cancer with distinct morphologies have been described in recent studies: polypoid growth type (PG-type) and non-polypoid growth type (NPG-type). We hypothesize that the morphologic differences may correspond to additional biological distinctions. Ratios of sialyl Lewisa (CA 19-9), sialyl Lewisx (SLX), or carcinoembryonic antigen (CEA) in the venous blood drainage from the tumor to that of the respective antigen in the peripheral venous blood (d/p ratio) was examined in order to ascertain whether or not the ratio is correlated with either the PG-type or NPG-type colorectal tumor growth pattern. Blood samples from 118 patients with colorectal cancer were obtained from a peripheral vein and from the tumor drainage vein during surgical excision of the tumor. Statistical tests were conducted by univariate and multivariate (logistic regression) analyses. Among the cancers examined there were 17 PG-type (14.4%) and 101 NPG-type (85.6%). NPG-type cancers had a higher frequency of moderately differentiated adenocarcinoma cells and T3/T4 tumors than PG-type cancers (P<0.0001 and P<0.0001, respectively). NPG-type cancers had a more advanced stage than PG-type cancers (P=0.0007). The d/p ratio of SLX in NPG-type cancers was significantly higher than that in PG-type cancers (P=0.028). Multivariate logistic regression analysis showed that three variables, namely histologic type, T factor, and d/p ratio of SLX, were independently related to tumor growth patterns. In conclusion, NPG-type cancers are characterized by a high SLX d/p ratio, which may be at least partly responsible for a different tumor progression pattern compared to other cancer types.

Topics: Adenocarcinoma; Aged; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Cell Division; Colonic Neoplasms; Colorectal Neoplasms; Female; Humans; Lewis Blood Group Antigens; Male; Middle Aged; Neoplasm Staging; Oligosaccharides; Rectal Neoplasms; Sialyl Lewis X Antigen; Veins

In this study, we examined the preoperative serum levels of sialyl Lewisa, sialyl LewisX, sialyl Tn, and carcinoembryonic antigen in 243 colorectal cancer patients in order to clarify the role of these antigens as prognostic factors after curative surgery. The patients were divided into two groups: low and high antigen groups (lower and higher than a selected diagnostic-based cut-off value). Patients with high serum levels of sialyl Lewisa and carcinoembryonic antigen had shorter disease-free intervals than those with low serum levels of the respective antigen, although sialyl Lewisx and sialyl Tn showed no significant differences. Multivariate analysis revealed that three independent prognostic variables, including depth of tumor invasion, lymph node metastasis, and serum sialyl Lewisa level, did prove to have value in predicting disease-free interval. In conclusion, among the four antigens examined in this study, the preoperative serum level of sialyl Lewisa is the only independent prognostic variable for recurrence after curative resection of colorectal cancer.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Colorectal Neoplasms; Disease-Free Survival; Female; Follow-Up Studies; Gangliosides; Humans; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Invasiveness; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen

Serum from patients with different malignancies contain an abnormal concentration of a a1-acidic-glycoprotein (AAG) and also, increased levels of AAG are associated with the presence of tumor mass. In the present report, serum levels of AAG were measured by radial immunodiffusion in squamous cell carcinoma of the head and neck (SCCHN) patients taking into account disease status parameters such as tumor localization, stage and extension of disease. Immunohistochemical methods, SDS-PAGE and Western-blotting were employed to study the expression of AAG and a carbohydrate related antigen (sialyl Lewis x) in tumor tissues and derived fractions. AAG showed abnormal levels in 7/15 oral cavity tumor patients sera, 2/5 oropharynx and 5/10 larynx tumors; increased AAG serum levels belonged to patients with disseminated disease. On the other hand, the presence of AAG and sialyl Lewis x were demonstrated in carcinoma cells and in derived fractions from tumor tissues belonging to patients with elevated AAG serum levels. In the present study, we have found elevated levels of AAG in serum samples from SCCHN patients; these neoplastic cells are capable to express AAG.

Topics: Adenocarcinoma; Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Carcinoma, Squamous Cell; Colorectal Neoplasms; Epithelial Cells; Female; Head and Neck Neoplasms; Humans; Male; Middle Aged; Neoplasm Metastasis; Neoplasm Proteins; Neoplasm Staging; Oligosaccharides; Organ Specificity; Orosomucoid; Sialyl Lewis X Antigen

Sialyl 6-sulfo Lewis X determinant has been described recently as a major ligand for L-selectin on high endothelial venules of human peripheral lymph nodes. From our investigation of its distribution in human colorectal cancer tissues and cultured colon cancer cells, the sialyl 6-sulfo Lewis X determinant was preferentially expressed in the nonmalignant colonic epithelia rather than cancer cells (P < 0.001; n = 23). This was in contrast to the distribution of conventional sialyl Lewis X, which was preferentially expressed in cancer tissues rather than nonmalignant epithelia (P = 0.007; n = 23), indicating that 6-sulfation predominantly occurs in nonmalignant tissues and is suppressed upon malignant transformation. In confirmation of this, a nonsialylated determinant 6-sulfo Lewis X was also found to be preferentially localized in the nonmalignant epithelia. Significant expression of sialyl 6-sulfo Lewis X was observed in only 2 lines, whereas 8 were positive for conventional sialyl Lewis X, among 13 cultured colon cancer cell lines. Transfection of cells with fucosyltransferase (Fuc-T) VI induced expression of sialyl 6-sulfo Lewis X, whereas transfection of Fuc-T III did not, suggesting that the determinant was synthesized mainly by Fuc-T VI in colonic epithelia. Members of the sialic acid cyclase pathway, the de-N-acetyl sialyl 6-sulfo Lewis X and cyclic sialyl 6-sulfo Lewis X determinants, were also preferentially expressed in the nonmalignant epithelia rather than colonic cancer cells (P < 0.001; n = 23). Stimulation of the sialyl 6-sulfo Lewis X-positive colon cancer cell line with a calcium ionophore ionomycin markedly reduced sialyl 6-sulfo Lewis X and induced cyclic sialyl 6-sulfo Lewis X expression. These results suggested that the metabolic conversion of sialyl 6-sulfo Lewis X into cyclic sialyl 6-sulfo Lewis X by a calcium-dependent enzyme, sialic acid cyclase, as we hypothesized for human leukocytes previously (C. Mitsuoka et al., Proc. Natl. Acad. Sci. USA, 96: 1597-1602, 1999), also occurs in nonmalignant colonic epithelia.

Topics: Colorectal Neoplasms; Humans; Immunohistochemistry; Lewis X Antigen; Ligands; Oligosaccharides; Sialyl Lewis X Antigen

Colorectal carcinoma can be morphologically divided into two different categories, namely polypoid growth (PG-type) and non-polypoid growth (NPG-type). To ascertain whether the expression of sialyl Le(x) antigen correlates with biologically and clinically important differences, an immunohistochemical assay was performed in 30 PG-type and 119 NPG-type cancers. In contrast to PG-type, the characteristics of the NPG-type include (1) an increased expression of sialyl Le(x); (2) a high rate of lymph node metastasis; (3) a high proportion of moderately differentiated adenocarcinoma cells; (4) young age of onset. It is concluded that differences in sialyl Le(x) expression between the PG-type and NPG-type cancers may be at least partly responsible for different tumor progression behavior.

Topics: Adenocarcinoma; Age of Onset; Aged; Cell Differentiation; Colorectal Neoplasms; Disease Progression; Female; Humans; Immunohistochemistry; Lewis X Antigen; Logistic Models; Lymphatic Metastasis; Male; Middle Aged; Multivariate Analysis; Neoplasm Staging; Oligosaccharides; Proportional Hazards Models; Sialyl Lewis X Antigen; Survival Rate; Up-Regulation

To investigate colorectal cancer-related carbohydrate antigen release and distribution, we evaluated serum levels of sialyl Le(a) (CA19-9) and sialyl Le(x) antigen (SLX) in blood samples obtained from both a peripheral vein and a tumor's draining vein.. Blood samples were obtained during surgery from 126 patients. Based on these samples, patients were placed into a high-antigen group, with a concentration above a selected cutoff value, or into a low-antigen group, with a tumor marker concentration below that same value. The blood samples obtained from peripheral veins were designated by the "p" prefix, and samples from drainage veins were designated by the "d.". Serum d-SLX levels were significantly higher than p-SLX levels (P < .0001), although there was no difference between those of d-CA19-9 and p-CA19-9. Only 1 (3.6%) of 28 patients in the high d-CA19-9 group had a low p-CA19-9. In contrast, 6 (33.3%) of 18 patients in the high d-SLX group had low p-SLX levels (P = .0103). Correlations between pathological variables and either p-CA19-9 levels or d-CA19-9 levels were similar. However, both distant metastasis and venous invasion did prove to be independent variables related to d-SLX levels, as shown by logistic regression analysis.. SLX may drain predominantly via the draining veins of colorectal tumors into portal circulation, whereas CA19-9 may drain via another route.

Topics: Aged; Antigens, Tumor-Associated, Carbohydrate; CA-19-9 Antigen; Colorectal Neoplasms; Female; Gangliosides; Humans; Logistic Models; Male; Middle Aged; Multivariate Analysis; Oligosaccharides; Sialyl Lewis X Antigen

Diets rich in omega-3 fatty acids have been shown to decrease both the initiation and promotion of colon carcinogenesis although their effect on hepatic metastasis formation is less well understood. Since adhesion of human colorectal carcinoma (HCRC) cells to hepatic endothelial cells is an important step in the metastatic cascade, the effect of membrane omega-3 fatty acid alterations on endothelial cell adhesion was studied.. CX-1 cells, a moderately differentiated HCRC cell line known to produce hepatic metastases in an athymic mouse intrasplenic injection model, were used. Cells were grown in omega-3 fatty acid-enriched medium and membrane-free fatty acid modifications confirmed with gas chromatography. Both human umbilical vein and hepatic sinusoidal endothelial cells were used in the binding assays. Adhesion assays were performed in a standard fashion using (51)Cr-labeled cells to tumor necrosis factor (TNF)-stimulated endothelial cell monolayers. Immunohistochemical analysis was performed for sialyl-Lewis(x), the receptor involved in endothelial adhesion on the surface of control and fatty acid-modified cells.. Gas chromatographic analysis confirmed membrane fatty acid modification of CX-1 cells by growth in docosahexanoic acid (omega-3) (4.761 nmol/10(6) cells vs 0.057 nmol/10(6) cells for controls). Binding of CX-1 to both human umbilical vein and hepatic sinusoidal endothelial cells decreased from 38.4 +/- 0.44 to 11.58 +/- 0.87% (P < 0.01). Immunocytochemical analysis showed a decrease in sialyl-Lewis(x) expression with omega-3 treatment.. These data indicate that omega-3 fatty acids may also be protective against the formation of hepatic metastases. The mechanism for this may be decreased endothelial cell adhesion which in turn may be due to decreased expression of the endothelial receptor sialyl-Lewis(x).

Topics: Animals; Cell Adhesion; Chromatography, Gas; Colorectal Neoplasms; Docosahexaenoic Acids; Endothelium, Vascular; Fatty Acids, Omega-3; Humans; Mice; Mice, Nude; Oligosaccharides; Sialyl Lewis X Antigen; Tumor Cells, Cultured; Umbilical Veins

Although the beneficial effect of cimetidine on survival in cancer has been clinically demonstrated in colorectal cancer patients, the mode of action of cimetidine has not been elucidated. In this report, we have demonstrated for the first time that cimetidine can block the adhesion of a colorectal tumor cell line to the endothelial cell monolayer in cell culture and that it can suppress the metastasis of the tumor cell in a nude mouse model. We also demonstrated that these antimetastasis effects of cimetidine might occur through down-regulation of the cell surface expression of E-selectin on endothelial cells, a ligand for sialyl Lewis antigens on tumor cells. We found that the cimetidine-mediated down-regulation of E-selectin did not involve down-regulation of E-selectin mRNA or blocking of the nuclear translocation of nuclear factor kappaB, a transcriptional activator of E-selectin gene expression. Because two other histamine type 2 receptor antagonists, famotidine and ranitidine, did not show any similar effect, these actions of cimetidine probably do not occur via blocking of the histamine receptor. These observations support the idea that cancer metastasis can be blocked by cimetidine administration through blocking the adhesion of tumor cells to the endothelium when an interaction between E-selectin and sialyl-Lewis antigens plays a role.

Topics: Animals; Cell Adhesion; Cell Nucleus; Cimetidine; Colorectal Neoplasms; Dose-Response Relationship, Drug; Down-Regulation; E-Selectin; Endothelium, Vascular; Enzyme Inhibitors; Enzyme-Linked Immunosorbent Assay; Famotidine; Histamine H2 Antagonists; Humans; Interleukin-1; Liver Neoplasms; Mice; Mice, Nude; Microscopy, Confocal; Neoplasm Transplantation; NF-kappa B; Oligosaccharides; Ranitidine; RNA, Messenger; Sialyl Lewis X Antigen; Transcriptional Activation; Tumor Cells, Cultured; Umbilical Veins

Subtraction values, (i.e. values obtained by subtracting the serum titer of sialyl Lewis(a) (CA19-9) and sialyl Lewis(x) (SLX) antigens in peripheral venous blood from the serum titer of the same antigen in the tumor's drainage venous blood) were determined in order to clarify whether or not such values for these specific antigens (d-CA19-9 and d-SLX) are prognostic factors after resection for colorectal cancer. The blood samples were obtained from 144 colorectal cancer patients during surgical excisions of the tumors. Univariate and multivariate analyses revealed that d-SLX level was an independent prognostic factor, separate from stage, while d-CA19-9 level did not have any additional prognostic value. In conclusion, a high d-SLX level is a predictor of poor outcome after surgery.

Topics: Adult; Aged; Aged, 80 and over; CA-19-9 Antigen; Colorectal Neoplasms; Female; Follow-Up Studies; Humans; Male; Middle Aged; Multivariate Analysis; Neoplasm Staging; Oligosaccharides; Prognosis; Sialyl Lewis X Antigen; Survival Analysis; Survival Rate

Recent studies delineated two different patterns of tumor growth in colorectal carcinoma characterized as polypoid and nonpolypoid (PG-type and NPG-type, respectively). We quantified serum sialyl Lewis (Le)a (CA19-9), sialyl Lex (SLX), sialyl Tn (STN), and carcinoembryonic antigen (CEA) in 269 colorectal cancer patients to establish whether their levels correlated with any biological or clinical differences between PG-type and NPG-type cancer. Patients were divided into high and low antigen groups (higher or lower than a selected diagnostic-based cut-off value) and compared. Statistical testing was by univariate and multivariate (logistic regression) analyses. Forty-seven (17.5%) patients with PG-type and 222 (82.5%) with NPG-type cancer were studied. In contrast to NPG-type, the characteristics of the PG-type cancers included a low rate of lymph node metastasis and a high serum STN level. In contrast to a low STN level, a high STN level was independently related to the presence of distant metastasis in patients with PG-type cancer, and also to the presence of distant metastasis and large-sized tumor in patients with NPG-type cancer. These data suggest that differences in STN levels in the serum of patients with PG-type or NPG-type colorectal carcinomas may be at least partly responsible for different tumor progression behavior.

Topics: Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Cell Division; Colorectal Neoplasms; Female; Gangliosides; Humans; Logistic Models; Male; Middle Aged; Multivariate Analysis; Neoplasm Staging; Oligosaccharides; Predictive Value of Tests; Sialyl Lewis X Antigen

Expression of mucin-bound sialyl-Le(x) antigen during the progression of colorectal carcinoma and its potential prognostic value were analysed in sections of tumours from 182 patients with a documented follow-up by immunohistochemistry using the monoclonal antibody (MAb) AM-3. Two groups of colonic carcinomas with weak (n = 79) and strong (n = 103) sialyl-Le(x) expression were discerned. The percentage of strongly expressing tumours increased with the progression of the disease (UICC stage I = 10%, stage II = 46%, stage III = 63%, stage IV = 68%, p < 0.0001). Seventy-four percent of patients with carcinomas exhibiting a strong sialyl-Le(x) expression but only 34% of patients with weak sialyl-Le(x) expression died of the disease (p = 0.0026). In multivariate analysis, strong sialyl-Le(x) expression increased the relative risk of cancer-related death 3.8-fold (95% CI = 1.8-7.9, p = 0.00034). The separate analyses of patients in UICC stage II (n = 56), III (n =5 9) and IV (n = 57) revealed that strong sialyl-Le(x) expression was associated with a reduction of the 5-year overall survival rate in UICC stage II (84% vs. 54%, p = 0.0013) and in stage III patients (86% vs. 35%, p = 0.0008) after curative resection but was not relevant in patients with distant metastases. In conclusion, the strong expression of sialyl-Le(x) antigen defined by the MAb AM-3 in colorectal carcinomas is an independent unfavourable prognostic factor after curative resection in stage II and III patients. The predictive power of the sialyl-Le(x) expression may be helpful to define subgroups of patients at high risk for whom preventive adjuvant therapy can be selectively applied before the occurrence of detectable metastases.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Colorectal Neoplasms; Disease-Free Survival; Female; Follow-Up Studies; Humans; Immunohistochemistry; Lewis Blood Group Antigens; Male; Middle Aged; Neoplasm Staging; Oligosaccharides; Predictive Value of Tests; Prognosis; Retrospective Studies; Sialyl Lewis X Antigen; Survival Rate; Time Factors

To study the correlation of sialyl Lewis-X (SLeX) antigen expression with the metastatic potential of human colorectal cancer.. The expression of biosynthetic enzyme of SLeX antigen--alpha1, 3Fuc-T mRNA in various cell lines of colorectal cancer possessed with different metastatic potentials (LoVo and HT(29)) was investigated by in situ hybridization. The expression of SLeX antigen was studied using immunohistochemistry and fluorescence-activated flow cytometry qualitatively and quantitatively.. The expression level of alpha1, 3Fuc-T mRNA and SLeX antigen in highly metastatic LoVo cells was high (21.2 +/- 7.7, 32.8 +/- 10.9, P < 0.05), and the expression level was low in HT(29) cells (10.8 +/- 5.2, 21.9 +/- 8.8) which are known possessed with a lower metastatic potential.. The expression of SLeX antigen is correlated to the metastatic potentiality of human colorectal cancer.

Topics: Cell Line, Tumor; Colorectal Neoplasms; Fucosyltransferases; HT29 Cells; Humans; Immunohistochemistry; Lewis X Antigen; Neoplasm Metastasis; Oligosaccharides; RNA, Messenger; Sialyl Lewis X Antigen

Two factors potentially determining the consistent overexpression of sialyl-Le(x) antigen in colon carcinoma and metastases were investigated: (i) the expression of the mucins MUC1 and MUC2, known to carry sialyl-Le(x), by Northern blotting; (ii) the extent of sialic acid O-acetylation, by Western blotting and HPLC. RNA and sialyl-Le(x)-positive mucins were purified from normal colonic mucosa (N), primary carcinomas (T) and their liver metastases (M). Northern blots showed that mRNA expression both of MUC1 and of MUC2 decreases during the progression of the disease, and is lowest in metastatic tissue. The expression of mucin-bound sialyl-Le(x) increased strongly from N to T and, to a lesser extent, to M. After alkali treatment of the mucins these differences disappeared, indicating that the total amount of mucin-bound sialyl-Le(x) is the same in the 3 types of tissues. The O-acetylation of mucin-bound sialyl-Le(x) gradually decreased from N to M. HPLC analysis showed that in N about 70%, in T 45% and in M only 20% of mucin-bound sialic acids are O-acetylated. Thus, the increase of sialyl-Le(x) detectable during colon-carcinoma progression is due to diminished O-acetylation and not to increased expression of mucin protein cores. The decrease of O-acetylation is therefore the primary chemical alteration contributing to colon carcinoma-associated overexpression of sialyl-Le(x).

Topics: Acetylation; Carcinoma; Colorectal Neoplasms; Humans; Liver Neoplasms; Mucin-1; Mucin-2; Mucins; Oligosaccharides; RNA, Messenger; Sialyl Lewis X Antigen

Recently, it was demonstrated that an increased level of NeuNAc alpha2-3Gal beta1-4(Fuc alpha1-3)GlcNAc beta-R (sialyl Le(x)) and NeuNAc alpha2-3Gal beta1-3(Fuc alpha1-4)GlcNAc beta-R (sialyl Le(a)) expression on the surface of colorectal cancer cells is positively correlated with progression of the disease. It has not been determined, however, which type of glycans, N- or O-glycans, is more closely associated with progression when cancer cells express those oligosaccharides. To address this problem, we have examined expression of sialyl Le(a) and sialyl Le(x), those oligosaccharides in O-glycans, and core 2 beta-1,6-N-acetylglucosaminyltransferase (C2GnT) transcripts in colorectal cancer specimens from 46 patients and compared those results with clinicopathological variables. C2GnT is a glycosyltransferase that is responsible for the core 2 branch, which is critical for biosynthesis of sialyl Le(a) and sialyl Le(x) in O-glycans. Sialyl Le(a) and sialyl Le(x) were determined by immunohistochemistry, and C2GnT transcripts were detected by reverse transcription-PCR. Sialyl Le(a) or sialyl Le(x) in O-glycans was assessed by combining immunohistochemistry for sialyl Le(a) or sialyl Le(x) with reverse transcription-PCR for C2GnT. Sialyl Le(a), detected on cancer cells in 74% of patients, was well correlated with lymph node metastasis, whereas sialyl Le(a) and sialyl Le(x) in O-glycans, which were specifically detected in cancer tissues of 50 and 61% of patients, respectively, were closely associated with lymphatic and venous invasion. In addition, C2GnT, which was specifically detected in cancer tissues of 63% of patients, was closely correlated with the vessel invasion, as well as depth of tumor invasion. These results strongly suggest that sialyl Le(a) and sialyl Le(x) in O-glycans and C2GnT, expressed in cancer cells, may play important roles in tumor progression through vessel or direct invasion.

Topics: Adult; Aged; Aged, 80 and over; Biomarkers, Tumor; Biopsy; CA-19-9 Antigen; Carbohydrate Sequence; Colorectal Neoplasms; Female; Gangliosides; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Lewis Blood Group Antigens; Lymphatic Metastasis; Male; Middle Aged; Molecular Sequence Data; N-Acetylglucosaminyltransferases; Neoplasm Invasiveness; Neoplasm Staging; Oligosaccharides; Polymerase Chain Reaction; Sialyl Lewis X Antigen; Transcription, Genetic

Ten human colorectal cancer (CRC) cell lines were implanted orthotopically into the ceca and also into the livers, muscles and peritoneal cavities of SCID mice in order to analyze the characteristics regulating metastatic behaviors of CRCs. All the CRC cell lines formed tumors in the muscle and cecum, but they could be classified into two groups: (1) six cell lines with high tumorigenicity in the liver (HTLs) forming differentiated tumors, and (2) four with no tumorigenicity in the liver (NTLs) forming poorly differentiated tumors in SCID mice. After orthotopic implantation, NTLs never metastasized to the liver, whereas HTLs did. Therefore, intrahepatic tumorigenicity and differential status were closely associated with liver metastasis whereas differentiation was not associated with lung metastasis. The 6 HTLs demonstrated an inverse correlation between liver metastases and peritoneal dissemination, and immunohistochemistry indicated expression of sLeX, CA19-9 and carcinoembryonic antigen in tumors which correlated well with the liver metastatic rate. We found a strong correlation between liver metastasis and intrahepatic tumorigenicity and could reproduce the clinical correlations between the pattern of the metastatic spread and the differentiation phenotype of CRC in vivo. We consider further examination using this model will be useful for analyzing the complex mechanisms involved in clinically metastasizing CRCs.

Topics: Adenocarcinoma; Animals; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Carcinogenicity Tests; Colorectal Neoplasms; Disease Models, Animal; ErbB Receptors; Humans; Injections, Intraperitoneal; Liver Neoplasms; Male; Mice; Mice, SCID; Neoplasm Transplantation; Oligosaccharides; Peritoneal Neoplasms; Sialyl Lewis X Antigen; Tumor Cells, Cultured

E-selectin is an adhesion molecule of endothelial cells that binds to cancer cells mediated by sialyl Lewis A (sLea) or sialyl Lewis X (sLe(x)). It is suspected to be involved in hematogenous metastasis of tumors. Therefore, it is worth examining E-selectin expression in human colorectal cancer and its hepatic metastasis. In the present study, E-selectin was clearly revealed on the endothelial cells of small vessels adjacent to cancer nests both in primary and in metastatic nests in immunohistochemistry. In these tissues, E-selectin was observed on the endothelial cells lining the lumen of small vessels. Its expression adjacent to cancer nests appears to be induced through some stimuli by cancer cells, since its degree of expression is inversely correlated to the distance of the blood vessels from the cancer nests (p < 0.001). Endothelial cells adjacent to the metastatic lesion expressed E-selectin more extensively than those adjacent to the primary foci. This is also in line with the finding on serum E-selectin levels which were significantly elevated in the metastatic group as compared with the non-metastatic group. The serum E-selectin level may provide useful information in the diagnosis for hepatic metastasis of colorectal cancer, although the results are still tentative.

Topics: Aged; Aged, 80 and over; CA-19-9 Antigen; Cell Adhesion Molecules; Colon; Colorectal Neoplasms; E-Selectin; Endothelium, Vascular; Female; Gangliosides; Humans; Immunohistochemistry; Liver; Liver Neoplasms; Male; Middle Aged; Oligosaccharides; Rectum; Sialyl Lewis X Antigen

The expression of the mucin-bound sialyl-Lewisx epitope is increased in the tissue of most colorectal carcinomas and in the sera of about 30% of tumor patients. In colon cancer, a portion of the sialyl-Lex groups detectable with the monoclonal antibody AM-3 is located on MUC1 (C. Hanski et al., Cancer Res., 53: 4082-4088, 1993). In order to characterize the major colon carcinoma-associated sialyl-Lex-positive glycoprotein components, the tissue- and serum-derived antigens were investigated. The buoyant densities of the sialyl-Lewisx-positive antigens from tumor and normal colonic tissues and from sera of patients with colon carcinoma and healthy donors correspond to that of mucins (1.40 g/ml). The sialyl-Lex-positive mucins purified from both tissues elute under nonreducing conditions in the void volume of a Sepharose CL-2B column, indicating a molecular mass more than 2 x 10(7) daltons. They yield in immunoblot after SDS gel electrophoresis under reducing conditions a main band at an apparent M(r) 880,000. Radioactive labeling revealed that the band at M(r) 880,000 is the major protein component in sialyl-Lewisx-positive mucins both from tumor and normal colonic tissue. In sera of colon carcinoma patients, the sialyl-Lex moiety is also detectable mainly on a M(r) 880,000 glycoprotein band and, additionally, on a M(r) 140,000 molecule as well as on alpha 1-acid glycoprotein. Sera from healthy donors exhibited only a sialyl-Lex-positive glycoprotein with the apparent M(r) 140,000. Sandwich ELISA as well as immunoblots of mucins purified from the colon carcinoma cell line LS174T indicated that the sialyl-Lex moiety migrating in the M(r) 880,000 band is located on MUC2 protein core. Together, these data suggest that sialyl-Lex antigen in colon, colon carcinoma, and the sera of patients with this tumor is located on the MUC2 molecule, consisting of several subunits with an apparent M(r) 880,000, linked via disulfide bridges. The increase of sialyl-Lex expression in colon carcinomas appears to be mainly due to a more frequent transfer of sialyl-Lex moieties onto the mucin core in tumor tissue.

Topics: Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Chromatography; Colon; Colonic Neoplasms; Colorectal Neoplasms; Electrophoresis; Humans; Immunoblotting; Molecular Weight; Mucins; Oligosaccharides; Sialyl Lewis X Antigen; Sodium Dodecyl Sulfate

Sialyl Lewis A (SLA) and sialyl Lewis X (SLX) have been shown to be specific ligands for endothelial leukocyte adhesion molecule-1 (ELAM-1), and may be involved in the process of adhesion between cancer cells and endothelium. We used immunohistochemical methods to study the expression of SLA, SLX and CEA in both primary tumors and matched metastatic liver lesions of colorectal carcinomas. Specimens from primary tumors and matched liver metastases from 24 patients with colorectal carcinomas were studied immunohistochemically. The degree of expression of CEA in liver metastases was similar to that in primary tumors, but SLA and SLX were expressed on a larger proportion of tumor cells in liver metastases than in primary tumors. Our findings suggest that colorectal carcinoma cells expressing SLA and/or SLX form metastatic liver tumors. They also suggest that expression of SLA and SLX in primary of colorectal carcinoma can be used as a prognostic indicator of metastasis.

Topics: Adult; Aged; Biomarkers, Tumor; CA-19-9 Antigen; Carcinoembryonic Antigen; Colorectal Neoplasms; Female; Gangliosides; Humans; Immunohistochemistry; Lewis Blood Group Antigens; Liver Neoplasms; Male; Middle Aged; Oligosaccharides; Sialyl Lewis X Antigen