lewis-x-antigen and Colorectal-Neoplasms

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

This review is concerned with the usefulness and the problem of biomarkers for cancer of digestive organs. Carcinoembryonic antigen (CEA) is a most popular and useful tumor marker for cancer of digestive organs. Squamous cell carcinoma (SCC) antigen and CYFRA have been reported as a useful tumor marker for esophageal cancer. CEA and CA 19-9 are a good prognostic factor in gastric cancer patients. The post-operative increase of serum CEA can be a predictive marker for the patients of colorectal cancer. Development of a radioimmunoassay for highly sensitive detection of tumor markers, they are considered to be useful for monitoring after treatment. But are not useful for the early diagnosis. The diagnosis of hepatocellular carcinoma (HCC) is based mainly on serological markers, such as alpha-fetoprotein and PIVKA-II. The two are useful complementary markers of HCC because they do not correlate with each other. But the problem of the false-positive rate for the patients with chronic hepatitis or liver cirrhosis is still remained. A typical marker of pancreatic and bile duct cancer is carbohydrate antigen, but the sensitivity of these markers is only 50%. Recent molecular biological analysis may be used as effective biomarkers in the diagnosis, prognosis, therapy, and risk assessment of digestive cancer.

Topics: alpha-Fetoproteins; Antigens, CD19; Antigens, Neoplasm; Biomarkers; Biomarkers, Tumor; Carcinoembryonic Antigen; Carcinoma, Hepatocellular; Carcinoma, Squamous Cell; Colorectal Neoplasms; Digestive System Neoplasms; Esophageal Neoplasms; Female; Humans; Keratin-19; Keratins; Lewis X Antigen; Liver Neoplasms; Pancreatic Neoplasms; Prognosis; Protein Precursors; Prothrombin; Stomach Neoplasms

Human colorectal carcinomas with increased metastatic potential and with poor prognosis are characterized by high content of sialyl-Lewis X carbohydrate antigen as detected by monoclonal antibody (mAb) FH6. The levels of these carbohydrate antigens apparently increase during colorectal carcinoma progression from nonmetastatic to metastatic tumors and inversely correlate with post operative survival of colon carcinoma patients. Cell lines have been selected for high and low levels of cell surface sialyl-Lewis X antigen using mAb FH6. The high expresser cells adhere strongly to cytokine-activated endothelial cells apparently through E-selectin. The high expresser cells are also adhesive to sections of human livers, although involvement of selectins in this process is unknown. Increased expression of sialyl-Lewis X antigens on the surface of these high expresser variant cells is apparently due to increased alpha 1,3-fucosyltransferase, a biosynthetic enzyme presumable responsible for the final step of the production of sialyl-Lewis X antigens. Human fucosyltransferase VI cDNA has been stably transfected into the low expresser variant cells. alpha 1,3-Fucosyltransferase activity, cell surface sialyl-Lewis X carbohydrate antigen, and adhesion to activated endothelial cells have been showed to increase in these transfectant cells. Furthermore, these cells show dramatic increase in their liver metastatic potential when it is tested by the intrasplenic injection into nude mice. We propose that carbohydrate antigens with unique peripheral epitopes serve as unique molecular phenotypes that determine colorectal cancer metastasis.

Topics: Animals; Antigens, Tumor-Associated, Carbohydrate; Colorectal Neoplasms; Humans; Lewis X Antigen; Mice; Mice, Nude; Neoplasm Invasiveness; Neoplasm Metastasis

To evaluate clinical importance of the expression of sialyl Lewis-X (sLe(x)) and sialyl Lewis-a antigen (sLe(a)) in gastrointestinal cancers, we examined immunohistochemically expression of the two antigens in esophageal, gastric, colorectal, and pancreatic cancer. Expression of sLe(x) and sLe(a) were associated with several clinicopathologic features which reflect tumor aggressiveness in esophageal, gastric and colorectal cancer, but not in pancreatic cancer. In esophageal and colorectal cancer, survival rate of the patients with sLe(x) positive tumors was significantly poorer than that of the patients with sLe(x) negative tumors, while in gastric cancer that with sLe(a) positive tumors was significantly poorer than that with sLe(a) negative. Cox's multivariate analysis revealed that sLe(x) expression status was one of the significant discriminants of prognosis in colorectal cancer patients and sLe(a) status in gastric cancer patients. These results suggest that sLe(x) and sLe(a) expression could be involved in aggressiveness of gastrointestinal cancer and might prove to be a potent marker for prognosis in patients with gastric cancer and colorectal cancer.

Topics: Adult; Aged; Antigens, Tumor-Associated, Carbohydrate; CA-19-9 Antigen; Carbohydrate Sequence; Colorectal Neoplasms; Female; Gangliosides; Gastrointestinal Neoplasms; Humans; Immunohistochemistry; Lewis X Antigen; Lymphatic Metastasis; Male; Middle Aged; Molecular Sequence Data; Multivariate Analysis; Neoplasm Invasiveness; Prognosis; Stomach Neoplasms

Four prognostics factors were investigated for colorectal cancer metastases. 1) There were statistically more venous invasions using Victoria blue elastic staining in patients with liver or lymph node metastasis than in those without metastasis. 2) The immunohistochemical expression rate of c-erbB-2 in liver metastasis cases was 27%, which was significantly higher than 3% in no metastasis cases. 3) Sialyl Lewis x (SLex) is related with cell adhesion. SLex positive rates in vessel invasion cancer cells were 71.4% with metastasis and 31.0% without metastasis. 4) Matrilysin is one of MMPs and it was significantly increased with Dukes stage by fluorescence intensity.

Topics: Colorectal Neoplasms; Humans; Lewis X Antigen; Liver Neoplasms; Lymphatic Metastasis; Matrix Metalloproteinase 7; Metalloendopeptidases; Neoplasm Invasiveness; Neoplasm Metastasis; Prognosis; Receptor, ErbB-2; Veins

Topics: Adenocarcinoma; Animals; Antigens, Neoplasm; Biomarkers, Tumor; Carbohydrate Sequence; Carbohydrates; Cell Adhesion; Colorectal Neoplasms; Humans; Lewis X Antigen; Liver Neoplasms; Mice; Molecular Sequence Data; Neoplasm Metastasis; Prognosis; Retrospective Studies; Splenic Neoplasms; Survival Analysis; Tumor Cells, Cultured

Human colorectal carcinomas with increased metastatic potential and with poor prognosis are characterized by the high content of sialyl-LeX carbohydrate antigens. The levels of these carbohydrate antigens apparently increase during colorectal carcinoma progression from non-metastatic to metastatic tumors. The levels of tumor-associated sialyl-LeX antigens are inversely correlated to the post surgical survival of colon carcinoma patients as revealed by retrospective studies. Cell lines selected for high levels of cell surface sialyl-LeX antigens metastasize to livers when they are injected intrasplenically into nude mice. The highly expressing cells also strongly adhere to activated endothelial cells apparently through E-selectin. We conclude that sialyl-LeX carbohydrate antigen is a unique molecular phenotype that determines colorectal cancer metastasis.

Topics: Cell Adhesion; Colorectal Neoplasms; Humans; Lewis X Antigen; Neoplasm Metastasis; Neoplasm Staging; Prognosis

Cancer metastasis is an ectopic growth of malignant cells. In human colorectal cancer, it is hypothesized that, during the progression of the disease to an advanced stage, highly malignant and metastatic tumor cells arise within primary tumors and become predominant. Based on this hypothesis, molecules associated with metastatic cells have been sought by the comparison of surgical specimens from patients at various clinical stages. Colorectal carcinomas with increased metastatic potential and with poor prognosis have been characterized by a loss of an organ-specific mucin determinant (sulfomucin), by an increased expression of non-intestinal sialomucins, and by an ectopic expression of adhesion ligands (sialyl-dimeric Lex antigens) on mucins.

Topics: Adenocarcinoma; Amino Acid Sequence; Base Sequence; Carbohydrate Sequence; Colorectal Neoplasms; Gene Expression Regulation, Neoplastic; Lewis X Antigen; Molecular Sequence Data; Mucins; Neoplasm Metastasis; Neoplasm Staging; Sialomucins

Potentiation of 5-fluorouracil/leucovorin (FUra/LV) cytotoxicity by IFN-gamma in colon carcinoma cells is dependent on FUra-induced DNA damage, the Fas death receptor, and independent of p53 and RNA-mediated FUra toxicity, which occurs in normal gastrointestinal tissues. This provides a rationale for enhancing the selective action of FUra/LV by IFN-gamma in the treatment of colorectal carcinoma. Based on results from our preclinical studies we designed a Phase I trial combining FUra (370 mg/m2) and LV (200 mg/m2), i.v. bolus daily x 5 days, with escalating doses of IFN-gamma (10-100 micro g/m2) s.c. on days 1, 3, and 5, every 28 days. Twenty-five patients with carcinomas were enrolled; 6 patients received IFN-gamma on days 1 and 3 only. The dose-limiting toxicity, stomatitis, occurred most frequently at 100 micro g/m2 IFN-gamma. Minor response or SD was observed in 2 of 9 patients and in 4 of 12 patients at dose levels of < or =50 micro g/m2 and > or =75 micro g/m2 IFN-gamma, respectively. Three evaluable chemonaive patients demonstrated partial response (2) or complete response (1). Serial plasma samples revealed peak FUra concentrations of >100 micro M; at 100 micro g/m2 IFN-gamma plasma concentrations >5 units/ml persisted for 6.5 h and >1 unit/ml for 28.5 h. The pharmacokinetic parameters of IFN-gamma correlated with a 2-3-fold up-regulation of Fas expression at 24 h in CD15+ cells in peripheral blood samples. Furthermore, clinically relevant IFN-gamma concentrations up-regulated Fas expression and sensitized HT29 colon carcinoma cells in vitro to FUra/LV cytotoxicity. On the basis of the modulation of Fas signaling, FUra/LV combined with IFN-gamma has shown activity in a Phase I trial in colorectal carcinoma and warrants additional evaluation in Phase II.

Topics: Adult; Aged; Aged, 80 and over; Antimetabolites, Antineoplastic; Antineoplastic Combined Chemotherapy Protocols; Cell Separation; Colorectal Neoplasms; Dose-Response Relationship, Drug; fas Receptor; Female; Flow Cytometry; Fluorouracil; Follow-Up Studies; Humans; Interferon-gamma; Leucovorin; Lewis X Antigen; Male; Middle Aged; Models, Biological; Signal Transduction; Time Factors; Tumor Cells, Cultured; Up-Regulation

Myeloid cells represent an abundant yet heterogeneous cell population in the colorectal cancer microenvironment, and their roles remain poorly understood.. We used multiplexed immunofluorescence combined with digital image analysis to identify CD14. Higher intraepithelial (. Myeloid cell populations occur in spatially distinct distributions and exhibit divergent, subset-specific prognostic significance in colorectal cancer, with mature CD14

Topics: Aged; Colorectal Neoplasms; Female; Fluorescent Antibody Technique; Granulocytes; HLA-DR Antigens; Humans; Lewis X Antigen; Lipopolysaccharide Receptors; Male; Microscopy, Fluorescence; Middle Aged; Monocytes; Phenotype; Predictive Value of Tests; Prognosis; Prospective Studies; Sialic Acid Binding Ig-like Lectin 3; Tumor Microenvironment; United States

Human microsatellite-stable (MSS) colorectal cancers (CRCs) are immunologically "cold" tumour subtypes characterized by reduced immune cytotoxicity. The molecular linkages between immune-resistance and human MSS CRC is not clear.. We used transcriptome profiling, in silico analysis, immunohistochemistry, western blot, RT-qPCR and immunofluorescence staining to characterize novel CRC immune biomarkers. The effects of selective antagonists were tested by in vitro assays of long term viability and analysis of kinase active forms using anti-phospho antibodies.. We identified the lymphocyte antigen 6 complex, locus G6D (LY6G6D) as significantly overexpressed (around 15-fold) in CRC when compared with its relatively low expression in other human solid tumours. LY6G6D up-regulation was predominant in MSS CRCs characterized by an enrichment of immune suppressive regulatory T-cells and a limited repertoire of PD-1/PD-L1 immune checkpoint receptors. Coexpression of LY6G6D and CD15 increases the risk of metastatic relapse in response to therapy. Both JAK-STAT5 and RAS-MEK-ERK cascades act in concert as key regulators of LY6G6D and Fucosyltransferase 4 (FUT4), which direct CD15-mediated immune-resistance. Momelotinib, an inhibitor of JAK1/JAK2, consistently abrogated the STAT5/LY6G6D axis in vitro, sensitizing MSS cancer cells with an intact JAK-STAT signaling, to efficiently respond to trametinib, a MEK inhibitor used in clinical setting. Notably, colon cancer cells can evade JAK2/JAK1-targeted therapy by a reversible shift of the RAS-MEK-ERK pathway activity, which explains the treatment failure of JAK1/2 inhibitors in refractory CRC.. Combined targeting of STAT5 and MAPK pathways has superior therapeutic effects on immune resistance. In addition, the new identified LY6G6D antigen is a promising molecular target for human MSS CRC.

Topics: B7-H1 Antigen; Benzamides; Cell Line, Tumor; Colorectal Neoplasms; DNA Mismatch Repair; Female; Fucosyltransferases; Gene Expression Regulation, Neoplastic; Humans; Immunoglobulins; Janus Kinase 1; Janus Kinase 2; Lewis X Antigen; Male; MAP Kinase Signaling System; Microsatellite Instability; Mitogen-Activated Protein Kinase Kinases; Programmed Cell Death 1 Receptor; Pyrimidines; STAT5 Transcription Factor; T-Lymphocytes

Metastasis is a multistep molecular network process, which is the major cause of death in patients with colorectal cancer (CRC). MicroRNAs (miRNAs) play pivotal roles in tumorigenesis as either tumor suppressors or oncogenes. Increased expression of fucosyltransferase4 (FUT4) has been reported to be associated with the invasive and metastatic properties of CRC. Here to identify potential key miRNAs and their target genes for colorectal cancer (CRC), we compared miRNA expression profiles between metastatic CRC cell SW620 and primary CRC cell SW480. Microarray analysis revealed that there were 85 differentially expressed miRNAs in SW620 cells with highly metastatic potential compared to SW480 cells with lowly metastatic potential. The expression of miR-26a and miR-26b were lower in SW620 cells than in SW480 cells, as well as downregulated in tumor tissues than in adjacent normal tissues of CRC patients. By applying bioinformatic approaches for the prediction of miRNA targeting 3'-UTR of FUT4, we identified FUT4 as one of the miR-26a/26b-targeted genes, while the expression of the target gene exhibited patterns opposite to that of miR-26a/26b in CRC cell lines, tumor tissues and corresponding adjacent tissues. Forced miR-26a/26b expression affected migratory behavior of CRC cells and FUT4 expression, while altered expression of FUT4 in CRC cell lines modulated progression upon transfection with miR-26a/26b mimic or inhibiter. FUT4 also regulated directly aggressiveness of SW620 and SW480 cells. Moreover, statistical analyses revealed that low miR-26a/26b levels and high expression of FUT4 were positively correlated with poor overall survival. The identified CRC-restricted miR-26a and miR-26b might be implicated in cancer progression via their target gene FUT4, suggesting their potential usage in CRC treatment.

Topics: Base Sequence; Carcinogenesis; Cell Line, Tumor; Colorectal Neoplasms; Down-Regulation; Female; Fucosyltransferases; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Genes, Tumor Suppressor; Humans; Lewis X Antigen; Male; MicroRNAs; Middle Aged; Neoplasm Invasiveness; Up-Regulation

The role of interleukin-17 (IL-17) in the tumor microenvironment is controversial. We analyzed the in situ tumor expression of IL-17 in colorectal cancer (CRC), adenoma and non-tumor tissue to explore the possible correlation of IL-17 expression to clinicopathological characteristics, tumor-infiltrating neutrophils (TINs) and survival in CRC.. We reviewed the records of 78 consecutive patients diagnosed with CRC. Archival tissues were used. Thirty-six patients with colorectal adenoma were also included. From the 78 CRC patients, we randomly chose 40 cases and collected non-tumor tissue at 10 cm from the edge of the resected tumor. Immunohistochemistry was performed using anti-IL-17 and anti-CD15 (targeting neutrophils) antibody, respectively. Real-time PCR was used to detect IL-17 mRNA in different tissues. Associations between IL-17 expression, clinicopathological parameters and prognosis were evaluated.. The level of IL-17 mRNA was higher in CRC than in adenoma and non-tumor tissue (P < 0.05). Positive IL-17 protein expression was observed more frequently in CRC as compared to colorectal adenoma and non-tumor tissue, respectively (P < 0.01). IL-17 expression correlated to well differentiation and early stage CRC. The number of CD15+ neutrophils significantly increased in CRC and positively correlated to the expression of IL-17 (P < 0.05). Both Kaplan–Meier analysis and multivariate Cox regression analysis indicated that patients with positive IL-17 expression showed better overall survival.. The association between IL-17 expression and the clinicopathological parameters, as well as the clinical outcome suggests a significant role of IL-17 in CRC. IL-17 is a marker of favorable prognosis.

Topics: Adenoma; Aged; Biomarkers, Tumor; Colorectal Neoplasms; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Interleukin-17; Kaplan-Meier Estimate; Lewis X Antigen; Male; Middle Aged; Neutrophils; Prognosis; Proportional Hazards Models; Real-Time Polymerase Chain Reaction; Treatment Outcome; Tumor Microenvironment

Cancer-related immune antigens in the tumor microenvironment could represent an obstacle to agents targeting EGFR "cetuximab" or VEGF "bevacizumab" in metastatic colorectal cancer (mCRC) patients.. Infiltrating immune cells into tumor tissues, cancer-related expression of immune antigens (CD3, CD8, CD68, CD73, MPO, CD15/FUT4) from 102 mCRC patients receiving first-line Cetuximab or Bevacizumab plus chemotherapy were assessed by immunohistochemistry and validated in an independent tissue microarrays of 140 patients. Genome-wide expression profiles from 436 patients and 60 colon cancer cell lines were investigated using bioinformatics analysis. In vitro kinase assays of target genes activated by chemokines or growth factors were performed.. Here, we report that cancer-related CD15/FUT4 is overexpressed in most of mCRCs patients (43 %) and associates with lower intratumoral CD3+ and CD8+ T cells, higher systemic inflammation (NLR at diagnosis >5) and poorer outcomes, in terms of response and progression-free survival than those CD15/FUT4-low or negative ones (adjusted hazard ratio (HR) = 2.92; 95 % CI = 1.86-4.41; P < 0.001). Overexpression of CD15/FUT4 is induced through RAF-MEK-ERK kinase cascade, suppressed by MEK inhibitors and exhibits a close connection with constitutive oncogenic signalling pathways that respond to ERBB3 or FGFR4 activation (P < 0.001). CD15/FUT4-high expressing colon cancer cells with primary resistance to cetuximab or bevacizumab are significantly more sensitive to MEK inhibitors than CD15/FUT4-low counterparts.. Cancer-related CD15/FUT4 overexpression participates in cetuximab or bevacizumab mechanisms of resistance in mCRC patients. CD15/FUT4 as a potential target of the antitumor immune response requires further evaluation in clinical studies.

Topics: Antineoplastic Agents; Antineoplastic Combined Chemotherapy Protocols; Bevacizumab; Biomarkers, Tumor; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Cetuximab; Cohort Studies; Colorectal Neoplasms; Disease-Free Survival; Drug Resistance, Neoplasm; ErbB Receptors; Extracellular Signal-Regulated MAP Kinases; Fucosyltransferases; Humans; Inflammation; Lewis X Antigen; Mitogen-Activated Protein Kinase Kinases; raf Kinases; Retrospective Studies; Tumor Microenvironment; Vascular Endothelial Growth Factor A

Colorectal cancer (CRC) is one of the most prevalent malignancies in Argentina with 11,043 new cases and 6,596 deaths estimated to have occurred in 2008. The present study was developed to clarify the differential expression of MUC1, MUC2, sLex, and sLea in colorectal cancer patients and their relationship with survival and clinical and histological features.. Ninety primary tumor samples and 43 metastatic lymph nodes from CRC patients were studied; follow-up was documented. Twenty-six adenoma and 68 histological normal mucosa specimens were analyzed. An immunohistochemical approach was applied and statistical analysis was performed.. In tumor samples, MUC1, sLea, and sLex were highly expressed (94%, 67%, and 91%, respectively); also, we found a significantly increased expression of the 3 antigens in primary tumors and metastatic lymph nodes compared with normal mucosa and adenomas. MUC2 was expressed in 52% of both normal mucosa and CRC samples; this reactivity significantly decreased in metastatic lymph nodes (p<0.05). A multiple comparison analysis showed that MUC1 and sLex discriminated among 3 groups: normal, adenoma, and CRC tissues. The increase of sLex expression showed an association with recurrence, and survival analysis showed that a high sLex staining was significantly associated with a poor survival. By multivariate analysis MUC1 inmunoreactivity correlated positively and significantly with tumor size, while MUC2 expression showed the opposite correlation.. The correlation of sLex overexpression in primary tumors and metastatic lymph nodes, the discrimination among the normal, adenoma, and CRC groups based on sLex expression, as well as its association with recurrence and survival, all suggest a prognostic role of sLex in Argentinian CRC patients.

Topics: Aged; Aged, 80 and over; Argentina; Biomarkers, Tumor; Colorectal Neoplasms; Female; Fucosyltransferases; Gene Expression Regulation, Neoplastic; Humans; Lewis X Antigen; Lymphatic Metastasis; Male; Middle Aged; Mucin-1; Neoplasm Recurrence, Local; Prognosis; Survival Analysis

Colorectal cancer (CRC) infiltration by adaptive immune system cells correlates with favorable prognosis. The role of the innate immune system is still debated. Here we addressed the prognostic impact of CRC infiltration by neutrophil granulocytes (NG).. A TMA including healthy mucosa and clinically annotated CRC specimens (n = 1491) was stained with MPO and CD15 specific antibodies. MPO+ and CD15+ positive immune cells were counted by three independent observers. Phenotypic profiles of CRC infiltrating MPO+ and CD15+ cells were validated by flow cytometry on cell suspensions derived from enzymatically digested surgical specimens. Survival analysis was performed by splitting randomized data in training and validation subsets.. MPO+ and CD15+ cell infiltration were significantly correlated (p<0.0001; r = 0.76). However, only high density of MPO+ cell infiltration was associated with significantly improved survival in training (P = 0.038) and validation (P = 0.002) sets. In multivariate analysis including T and N stage, vascular invasion, tumor border configuration and microsatellite instability status, MPO+ cell infiltration proved an independent prognostic marker overall (P = 0.004; HR = 0.65; CI:±0.15) and in both training (P = 0.048) and validation (P = 0.036) sets. Flow-cytometry analysis of CRC cell suspensions derived from clinical specimens showed that while MPO+ cells were largely CD15+/CD66b+, sizeable percentages of CD15+ and CD66b+ cells were MPO-.. High density MPO+ cell infiltration is a novel independent favorable prognostic factor in CRC.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Antigens, Differentiation, Myelomonocytic; Cell Adhesion Molecules; Colorectal Neoplasms; Female; Flow Cytometry; Fucosyltransferases; GPI-Linked Proteins; Humans; Immunohistochemistry; Lewis X Antigen; Male; Middle Aged; Multivariate Analysis; Neoplasm Grading; Neoplasm Staging; Neutrophils; Peroxidase; Prognosis; Receptors, IgG; Survival Analysis

Prediction of poor patient outcome as a effect of adjuvant therapy in stage II colorectal cancer (CRC) remains a matter of controversy. Tumor expression of CD10 and CD15 is reportedly related to poor outcome in CRC. In this study, we investigated whether the expression of CD10 and CD15 is a predictor of outcome and the effect of adjuvant therapy in stage II CRC.. Immunohistochemical analyses for CD10 and CD15 and some additional markers (CD11b, CD14, CD163, CD3, and CD20) were performed using paraffin sections of CRC specimens from 57 patients who had undergone curative surgical treatments between 1998 and 2004. Forty of these patients received postoperative adjuvant therapy. We distinguished between expression in tumor cells (tCD10 and tCD15), in stromal cells (sCD10), and infiltrating immune cells (iCD10 and iCD15).. Expression of iCD10 was observed in 21.1% (12/57) of the specimens examined. Of all expression patterns, only iCD10 expression at the cancer invasion front was a useful predictor of a disease-free period and overall survival in stage II CRC. Adjuvant therapy improved the outcome of iCD10(+) patients. CD10(+) immune cells were heterogeneous in origin and partially overlapped the cells positive for myeloid lineage markers, including CD11b and CD15.. iCD10 expression at the tumor invasion front is a useful marker for predicting a high risk of recurrence and mortality in stage II CRCs. CD10(+) immune cells appear to be of myeloid origin.

Topics: Adult; Chemotherapy, Adjuvant; Colorectal Neoplasms; Fucosyltransferases; Humans; Lewis X Antigen; Male; Middle Aged; Myeloid Cells; Neoplasm Invasiveness; Neoplasm Staging; Neprilysin; Prognosis; Treatment Outcome

Members of the family of carcinoembryonic antigen (CEA)-related cell adhesion molecules (CEACAMs) belonging to the immunoglobulin (Ig) superfamily are expressed in a variety of normal and malignant human tissues. As components of the cell membrane, these glycoproteins can make contact with adjacent cells. CEACAM1 and CEACAM5 (CEA) express Lewis(x) (Le(x)) structures. As shown by mass spectrometry in conjunction with enzymatic digestion, CEACAM1 contains at least seven Le(x) residues. Fucosyltransferase IX is the main fucosyltransferase responsible for attachment of terminal fucose, the key feature of the Le(x) structure, to CEA and CEACAM1. The Le(x) residues of both, CEACAM1 and CEA, interact with the human Le(x)-binding glycan receptors DC-SIGN and SRCL. Since subpopulations of human macrophages express DC-SIGN or SRCL, Le(x)-carrying CEACAMs may modulate the immune response in normal tissues such as the human placenta or in malignant tumours, for example in colorectal, pancreatic or lung carcinomas.

Topics: Antigens, CD; Carcinoembryonic Antigen; Cell Adhesion Molecules; Cell Line; Collectins; Colorectal Neoplasms; Female; Fucose; Fucosyltransferases; Humans; Intestinal Mucosa; Lectins, C-Type; Lewis X Antigen; Melanoma; Placenta; Polysaccharides; Pregnancy; Protein Binding; Receptors, Cell Surface; Receptors, Scavenger; Recombinant Proteins; Skin Neoplasms; Tissue Extracts

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

Dendritic cells play a pivotal role in the induction of antitumor immune responses. Immature dendritic cells are located intratumorally within colorectal cancer and intimately interact with tumor cells, whereas mature dendritic cells are present peripheral to the tumor. The majority of colorectal cancers overexpress carcinoembryonic antigen (CEA), and malignant transformation changes the glycosylation of CEA on colon epithelial cells, resulting in higher levels of Lewis(x) and de novo expression of Lewis(y) on tumor-associated CEA. Dendritic cells express the C-type lectin dendritic cell-specific intercellular adhesion molecule-3-grabbing nonintegrin (DC-SIGN) that has high affinity for nonsialylated Lewis antigens, so we hypothesized that DC-SIGN is involved in recognition of colorectal cancer cells by dendritic cells. We show that immature dendritic cells within colorectal cancer express DC-SIGN and that immature dendritic cells but not mature dendritic cells interact with tumor cells. DC-SIGN mediates these interactions through binding of Lewis(x) and Lewis(y) carbohydrates on CEA of colorectal cancer cells. In contrast, DC-SIGN does not bind CEA expressed on normal colon epithelium that contains low levels of Lewis antigens. This indicates that dendritic cells may recognize colorectal cancer cells through binding of DC-SIGN to tumor-specific glycosylation on CEA. Similar to pathogens that target DC-SIGN to escape immunosurveillance, tumor cells may interact with DC-SIGN to suppress dendritic cell functions.

Topics: Antigens, CD; Antigens, Differentiation; Breast Neoplasms; Carcinoembryonic Antigen; Cell Adhesion Molecules; Cell Line, Tumor; Colorectal Neoplasms; Dendritic Cells; Glycosylation; Humans; Immune Tolerance; Lectins, C-Type; Lewis Blood Group Antigens; Lewis X Antigen; Receptors, Cell Surface

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

To gain new insights into the functional interaction between DC and neoplastic cells, we have analyzed the effects of melanoma and colorectal cancer lines on the chemotaxis and the phenotype of monocyte-derived DC in vitro. Both types of tumor cells displayed effective chemoattractive capacity towards immature, but not mature DC. Furthermore, conditioned medium of discrete melanoma lines induced upregulation of CD80, CD86, MHC class I, and MHC class II molecules on immature DC. However, de novo expression of E-cadherin and strong upregulation of CD15 could also be detected in the absence of CD83 expression. Melanoma-conditioned DC exhibited an increased adhesion capacity to a melanoma cell line in vitro and did not migrate in response to SLC chemokine. Tumor-infiltrating CD15(+) cells displaying DC morphology could also be detected by immunohistochemistry in the original tumor specimens from which discrete melanoma cell lines under investigation were derived. Colorectal cancer cell lines, although able to chemoattract immature DC, were apparently unable to modulate their phenotype. Altogether our results suggest that tumor cells can attract immature DC in vitro and, eventually, modulate their phenotype. As a result, DC mobility could be severely impaired.

Topics: Animals; Cadherins; Chemokines; Chemotaxis, Leukocyte; Colorectal Neoplasms; Culture Media, Conditioned; Dendritic Cells; Gene Expression Regulation, Neoplastic; Humans; Immunohistochemistry; Immunophenotyping; Lewis X Antigen; Melanoma; Mice; Tumor Cells, Cultured; Up-Regulation

Topics: Animals; Antibodies, Monoclonal; Cell Adhesion; Colorectal Neoplasms; E-Selectin; Endothelium, Vascular; Flow Cytometry; HT29 Cells; Humans; Immunohistochemistry; In Situ Hybridization; Lewis X Antigen; Liver Neoplasms; Mice; Mice, Inbred BALB C; Mice, Nude; Microscopy, Electron; Microscopy, Electron, Scanning; N-Acetylneuraminic Acid; RNA, Messenger; Tumor Cells, Cultured; Umbilical Veins

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 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

Various tumor-associated antigens have been identified as carbohydrates bound to lipids or to proteins expressed on tumor cell membranes. We prepared tumor-specific immunoliposomes by coupling anticarbohydrate antibodies, such as antiganglioside G(M3) antibody (DH2) or anti-Le(x) antibody (SH1), to polyethylene glycol (PEG)-coated liposomes. In vitro and in vivo targetability of anti-G(M3) and anti-Le(x) immunoliposomes to B16BL6 mouse melanoma cells and HRT-18 human colorectal adenocarcinoma cells were monitored with a fluorescence microscopy, and analyzed by biodistribution assay of the immunoliposome in mice bearing the tumor tissues. The antibody coupling to the PEG liposomes did not greatly diminish the circulation time of the liposome in the C57BL/6 mouse model. In vitro cytotoxicity of doxorubicin encapsulated in liposomes was enhanced by antibody coupling, but still behind free doxorubicin. However, in vivo antitumor therapeutic efficacy of doxorubicin encapsulated in the immunoliposomes was far greater than the free drug or in conventional liposomes. Doxorubicin encapsulated in anti-G(M3) immunoliposomes was able to reduce in vivo tumor growth and metastasis of B16BL6 mouse melanoma cells more greatly than any other formulations of the drug. This study suggests that tumor-associated antigens can be good target molecules for tumor-specific delivery of liposomal drugs or other synthetic drug delivery systems.

Topics: Animals; Antibodies, Monoclonal; Antigens, Neoplasm; Antineoplastic Agents; Colorectal Neoplasms; Doxorubicin; Drug Carriers; G(M3) Ganglioside; Humans; Immunotoxins; Lewis X Antigen; Liposomes; Melanoma, Experimental; Mice; Tumor Cells, Cultured

Recognition of metastatic tumor cells with distinct biochemical phenotypes predominant in the primary tumors should be useful not only for establishment of new therapeutic approaches but also for identification of high-risk or low-risk patients for relapse. We examined whether carbohydrate antigens, sialyl Lewis(x) (sLe(x)) and sialyl Lewis(a) (sLe(a)) are involved in colorectal cancer metastasis.. Metastatic abilities of human colon cancer cell variants that were selected for their high or low cell surface levels of sLe(x) (KM12-HX and KM12-LX, respectively) were analyzed. Also, immunohistochemical expressions of sLe(x) and sLe(a) in 159 primary colorectal cancers were examined to determine the clinical significance of increased expression of these antigens.. KM12-HX cells adhered more readily to tumor necrosis factor-alpha activated endothelial cells than did KM12-LX cells. Increased adhesion of KM12-HX cells to activated endothelial cells was inhibited by antibodies against E-selectin and sLe(x) and by modification of cell surface carbohydrates. KM12-HX cells showed more invasive ability in vitro and more metastatic potential in the liver of nude mice than KM12-LX cells. Although no difference was seen in the expression of six messenger ribonucleic acids corresponding to progression or metastasis of colorectal cancer, expression of fucosyltransferase was found to be responsible for the higher expression of sLe(x) in KM12-HX cells. Clinical records of patients showed that disease-free survival rate of patients with sLe(x)-positive tumors was significantly poorer than that of those with sLe(x)-negative tumors. Cox's multivariate analysis revealed that the sLe(x) status was an independent predictive factor for disease recurrence (P = 0.004), depth of invasion (P = 0.0005), and histologic type (P = 0.037), but sLe(a) status, age, gender, tumor location, N stage, and vessel invasion were not.. Increased expression of sLe(x) could be involved in establishment of colorectal cancer metastasis. It appears that examining sLe(x) expression may serve as a potent marker of the recurrence in patients with colorectal cancer.

Topics: Adult; Aged; Aged, 80 and over; Animals; Biomarkers, Tumor; Caco-2 Cells; Colorectal Neoplasms; Disease-Free Survival; Female; Humans; Immunohistochemistry; Lewis X Antigen; Liver Neoplasms, Experimental; Male; Mice; Mice, Inbred BALB C; Middle Aged; Neoplasm Recurrence, Local; Predictive Value of Tests; Prognosis; Proportional Hazards Models

The rate of polarized secretion of a putative adhesion ligand, sialosyl Lewis(a) (19-9), by SW1116 colorectal carcinoma cells is stimulated at least 20-fold after pre-incubation with, and the incorporation of, retinoic acid (RA). In order to investigate the possible involvement of fatty acylation in the export of the epitope, purified ligands from carcinoma-cell membranes, membrane subfractions and media were analyzed during RA-induced secretion. Incorporation of radioactivity from (3H)palmitate into membrane subfractions and purified sialosyl Lewis(a) antigenic molecular species of M(r) > 150,000 (SiaLeams) was stimulated by RA treatment. Most of the intracellular lipid radioactivity which bound to solid-phase 19-9 antibody behaved chromatographically, either like ganglioside or like NH2 OH-labile acyl groups, but most of the (3H) bound to SiaLeams of post-incubation media behaved like base-labile fatty acyl groups, or free fatty acid. Release of base-labile lipid radioactivity after 3 hr (associated with antigen) was almost exclusively into the apical media of membrane inserts. Gas-liquid chromatography/mass spec. analyses of purified Sialeams revealed the presence of palmitate (16:0), as well as stearate (18:0) and oleate (18:1) fatty acyl groups. Our results suggest that fatty acylation of SiaLeams may be co-ordinated with alterations in glycosylation and participate in directing these molecules to the apical surface. Lipid analyses were consistent with ganglioside chaperonage of SiaLeams to the apical surface, where N-fatty-acylated gangliosides remain for the most part integrated into the bilayer, but some oxyester or thioester bonds may be cleaved to permit release of SiaLeams to the apical medium.

Topics: Acylation; Chromatography, Gas; Colorectal Neoplasms; Electrophoresis, Polyacrylamide Gel; Fatty Acids; Fluorescent Antibody Technique; Glycoproteins; Humans; Lewis X Antigen; Mass Spectrometry; Palmitic Acid; Palmitic Acids; Tretinoin; Tumor Cells, Cultured

The immunohistochemical analysis using the monoclonal antibodies to the blood group antigens was performed on the tissues of 387 colorectal cancers and 307 normal colonic tissues, and the relation between expression of these antigens and prognosis of the patients with stage 2 was also investigated. La, Le(b), and CA 19-9 were used for type 1 antigens, and Le(x), Le(y), and Sialyl Le(x) for type 2, respectively. Expression of Ca 19-9 and Silyl Le(x) remarkably increased in cancers throughout the colorectum. Le(b) and Le(y) were expressed in only 0-17% of the normal distal colon, but in almost colonic tumors. Therefore CA 19-9 and Sialyl Le(x) are tumor-associated antigens in the whole colorectum, and Leb and Le(y) in the distal colon. Expression of CA 19-9 in primary tumor provided more prognostic information and risk of liver metastasis than other Lewis blood group antigens.

Topics: Adult; Aged; Aged, 80 and over; Antigens, Tumor-Associated, Carbohydrate; CA-19-9 Antigen; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Lewis Blood Group Antigens; Lewis X Antigen; Male; Middle Aged; Prognosis

The immunohistochemical expression of sialylated and non-sialylated forms of both Le(x) and Le(a) were studied in 87 carcinomas and 42 normal mucosal specimens of colon and rectum, as well as in 32 metastatic lymph nodes and 9 hepatic lesions, using an indirect immunoperoxidase staining. Their antigens were expressed in normal mucosa with the following frequencies: Le(a), 95.2% (40/42); sialyl Le(a), 88.1% (37/42); Le(x), 95.2% (40/42); and sialyl Le(x), 17.0% (7/42), whereas in carcinomas, the respective rate of frequency were: 78.2% (68/87); 78.2% (68/87); 90.8% (79/87); and 93.1% (81/87). Sialyl Le(x) antigen showed the highest tumor specificity compared to other antigens. In three normal mucosal specimens and four carcinomas with Le(a-b-) phenotype, the expression of type 1 antigens (Le(a) and sialyl Le(a)) was not consistent, whereas type 2 antigens (Le(x) and sialyl Le(x)) were consistently observed in carcinomas. The staining of type 1 antigens and Le(x) was decreased in metastatic lesions compared with primary carcinomas, whereas sialyl Le(x) antigen had the same positive-staining rate in both. Metastatic carcinoma expressed the sialylated form more predominantly than the non-sialylated form in type 2 antigens whereas the opposite result was observed in type 1 antigens. These results suggested that: (a) sialyl Le(x), defined by monoclonal antibody CSLEX1, may be useful as a tumor-associated antigen in colorectal carcinoma, and (b) the alteration of Lewis-related carbohydrate antigens in cancer cell membranes, including sialylation and/or aberrant glycosylation, may be related to metastatic behavior.

Topics: Adenocarcinoma; Adenocarcinoma, Mucinous; Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Colorectal Neoplasms; Glycosylation; Humans; Immunohistochemistry; Lewis Blood Group Antigens; Lewis X Antigen; Liver Neoplasms; Lymphatic Metastasis

Highly malignant and metastatic tumor cells are thought to arise within primary tumors and become predominant during cancer progression. We demonstrated, by the analysis of > 500 surgical specimens, that colorectal carcinomas with increased metastatic potential were characterized by an increased expression of sialyl-Le(x) antigens expressed on mucins. The biological role of sialyl-Le(x) antigens expressed on mucins produced by colon carcinoma cells has been investigated using variant cell lines selected for their expression of this antigen. KM12-HX and KM12-LX, high and low expresser variant cells, differed in their metastatic potential in nude mice after intrasplenic injection. KM12-HX cells contain higher levels of polyA+mRNA for alpha(1-3/4) fucosyltransferase than KM12-LX cells. Sialyl-Le(x) antigenic carbohydrate chains were attached to mucins as well as glycoproteins with various M(r). KM12-HX cells adhered more strongly than KM12-LX cells to human umbilical vein endothelial cells treated with tumor necrosis factor-alpha and to mouse hepatic sinusoidal endothelial cells. We have retrospectively evaluated post-operative survival of colon carcinoma patients for their sialyl-Le(x) antigen levels in the primary tumors according to the percentage of stained cells by specific antibodies. The adjusted survival rate of the patients with high levels of sialyl-Le(x) antigen in their primary tumors was much lower due to recurrence and metastasis than that of the patients with tumors containing low levels of sialyl-Le(x) antigen. The results suggested that sialyl-Le(x) antigen has a potential to be used as a predictive marker for colorectal cancer metastasis.

Topics: Animals; Antibodies, Monoclonal; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Cell Adhesion; Colorectal Neoplasms; Fucosyltransferases; Humans; Lewis X Antigen; Mice; Mice, Nude; Neoplasm Metastasis; Prognosis

We have previously shown that sialyl Lewisx antigen (sLex) (NeuAc alpha 2-3Gal beta 1-4(Fuc alpha 1-3)GlcNAC-R) has an important functional role in defining the invasion and metastasis of human colorectal carcinoma. The results were derived from the clinical specimens obtained at surgery or experimental metastasis of human colon carcinoma variant expressing different levels of sLex in nude mice. In the present study, we immunohistochemically examined 132 human colorectal carcinomas for the expression of sLex to investigate whether this antigen expression could serve as a prognostic parameter. The tumors were divided into two groups: sLex positive and sLex negative. The incidence of sLex positive was correlated with the depth of tumor invasion, the presence of the lymph node metastasis, lymphatic invasion, and the disease stage. The difference was statistically significant (P = 0.0026; P = 0.0002; P = 0.003; P = 0.0013; respectively). Based on the data on 114 patients who underwent curative resections, incidence of the disease recurrence was assessed. The sLex-positive patients had higher incidence of recurrence in distant organs, especially in the liver, than that of the sLex-negative patients. The 5-year disease free survival rates of sLex-positive and -negative patients were 57.7 and 89.1%, respectively (P = 0.0002). The difference of 5-year overall survival rates between the two were also significant (sLex positive, 58.3%; sLex negative, 93.0%: P < 0.0001). By Cox multivariate analysis, sLex expression levels remained the best discriminant of disease-free survival (P = 0.035) and overall survival (P = 0.0081). These results suggest that increased expression of sLex is correlated with the extent of malignancy and high incidence of recurrence and consequently with survival of colorectal carcinoma patients. Thus sLex may prove to be a potent marker of recurrence in colorectal carcinoma patients.

Topics: Adult; Aged; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Lewis X Antigen; Male; Middle Aged; Survival Rate

Pokeweed mitogen (PWM) lectin, known to bind branched poly-N-acetyllactosamines, has a highly selective affinity for human colorectal carcinomas. We performed light microscopic (LM) histochemistry with PWM lectin on paraffin sections of human colorectal tissues. In histological sections, normal mucosae and adenomas with mild dysplasia exhibited negative reaction (0/10, 0/13, respectively) with or without neuraminidase pre-digestion, whereas adenomas with moderate dysplasia showed a small increase in PWM lectin reactivity after neuraminidase digestion (4/23). In contrast, we observed a high incidence of positive reactivity in colorectal carcinoma without neuraminidase pre-digestion (38/44). After digestion with neuraminidase, there was increased reactivity of colorectal carcinomas in situ (7/12) and invasive carcinomas (13/32). These results imply that human colorectal carcinomas consistently contain substantial amounts of PWM-reactive branched poly-N-acetyllactosamine glycoconjugates structures. We also compared the staining patterns of PWM lectin and monoclonal antibodies (MAb) directed to Lewis X (LeX) or Lewis Y (LeY) antigen. PWM lectin reactivity was largely confined to the apical membranes of carcinoma tissues. MAb-LeX or MAb-LeY immunoreactivity was seen on the apical membranes and in the cytoplasm of both adenomas and carcinomas. Therefore, histochemical studies with this lectin should be useful for identification of carcinoma tissues and analysis of glycoconjugates associated with colorectal carcinoma.

Topics: Adenoma; Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Carcinoma in Situ; Colorectal Neoplasms; Female; Glycoconjugates; Histocytochemistry; Humans; Immunohistochemistry; Intestinal Mucosa; Lewis X Antigen; Male; Middle Aged; Pokeweed Mitogens

A hepatic invasive human colorectal xenograft model was derived in nude mice by selection through the liver of the parental cell line, C170. Following intraperitoneal injection, tumours selectively grew on the liver in > 80% of the animals within 15-20 days. The liver-invading xenograft line, renamed C170HM2, had a significantly greater expression of the Lewisx antigen compared to C170 (mean linear fluorescence per cell > 1000 compared with 500 for C170, P < 0.02). C170HM2 had significantly elevated proliferation (when compared with C170) in the presence of epidermal (P < 0.001) and basic fibroblast growth factor (P < 0.001). C170HM2 also mitogenically responded to type I collagen (derived from rat tails), unlike C170. C170HM2 tumours when invading the liver expressed both interstitial collagenase and gelatinase activity at the invading edge.

Topics: Animals; Antigens, Neoplasm; Carcinoembryonic Antigen; Collagenases; Colorectal Neoplasms; Disease Models, Animal; Fibroblast Growth Factor 2; Gangliosides; Lewis X Antigen; Liver Neoplasms, Experimental; Male; Mice; Mice, Nude; Mitosis; Neoplasm Invasiveness; Neoplasm Transplantation

The alterations of carbohydrate chains in cancer cell membrane can be related, not only to the formation of tumor-associated antigens, but also to cell biologic significance. There is, furthermore, a possibility of their relationship to tumor metastatic behavior and subsequent survival of patients with cancer. Recent clinical studies elucidated that a carbohydrate antigen, sialyl Lex, is a useful tumor marker in colorectal cancer. However, the sialyl Lex antigen immunoreactivity in colorectal carcinoma in relation to patient survival is unknown. The aim of the study was to elucidate whether sialyl Lex expression in tumors was correlated with patient survival.. Immunohistochemical expression of sialyl Lex antigen, as detected by monoclonal antibody CSLEX1, was studied in 175 specimens of primary colorectal carcinoma from 120 patients who received radical surgery.. The positive expression of sialyl Lex was observed in 87 of 120 patients (72.5%). There was no statistically significant association between the negative or positive of sialyl Lex and clinicopathologic factors, excluding Dukes stage and histologic type. The difference between overall 5-year survival of patients with sialyl Lex-negative tumors and that of patients with sialyl Lex-positive tumors (81.2% versus 60.6%) was statistically significant (P = 0.0263). In proportion to staining intensity that was expressed as a score based on the percentage of the total field stained positive with CSLEX1, 5-year survival of patients indicated a worse outcome (P = 0.0113). The prognostic value was then studied in a Cox regression model. Dukes stage had the strongest association with patient survival, whereas sialyl Lex expression was found to be the second-ranking parameter.. When examining the expression of sialyl Lex antigen, as detected by monoclonal antibody CSLEX1, clinically useful information for patient survival after radical resection of colorectal cancer is given.

Topics: Adenocarcinoma; Aged; Antibodies, Monoclonal; Colorectal Neoplasms; Female; Humans; Immunohistochemistry; Lewis X Antigen; Male; Middle Aged; Multivariate Analysis; Neoplasm Staging; Prognosis; Proportional Hazards Models; Survival Rate

The immunohistochemical analysis using the monoclonal antibodies to the blood group associated antigens Lea, Leb, CA19-9, and CEA, were performed on the tissues of the colorectal cancers, adenomas, normal mucosae and distant metastatic lesions, to determine whether the given degree of expression have some relation to the prognosis of patients or the potential of metastasis. We also analysed the serum levels of CA19-9 and CEA by RIA and compared them with the results of immunopathology. The more pronounced expression of CEA, Lea, Leb, and CA19-9 was observed in proportion to the progress of the cancer. In those cases of immunohistochemically Lea negative results in tissues, CA19-9 was not detected either in the serum level nor in tissue sections, therefore all cases of Lea negative results were eliminated in the evaluation of the clinical significance of CA19-9. Positive expression of CA19-9 in tissue of cancer indicated a poor prognosis of the patient and high risk of distant metastasis. Higher intensity of staining was observed in hepatic metastasis, in comparison with the primary lesion. These studies demonstrated that immunohistochemical analysis of CA19-9 and blood group antigens in useful tool for evaluation of prognosis and risk of metastasis in colorectal cancer.

Topics: Adult; Aged; Carcinoembryonic Antigen; Colorectal Neoplasms; Female; Humans; Lewis X Antigen; Lymphatic Metastasis; Male; Middle Aged; Neoplasm Metastasis; Neoplasm Staging; Prognosis

Sialyl-dimeric LeX antigen (NeuAc alpha 2-3Gal beta 1-4[Fuc-alpha 1-3] GlcNAc beta 1-3Gal beta 1-4[Fuc alpha 1-3]GlcNAc beta 1-R; SLX) is an oncodevelopmental carbohydrate antigen expressed both on glycolipids and on mucin-like glycoproteins in human colorectal carcinomas. The levels are higher in primary tumors at advanced stages than in tumors at early stages, and metastatic lesions contain a greater quantity of this antigen than corresponding primary tumors. To study whether this antigen influences the metastatic behavior of tumor cells, we selected SLX variant cells from the human colon carcinoma cell line KM12C using the specific monoclonal antibody FH6. We obtained two stable cell lines: a high expresser (KM12-HX) and a low expresser (KM12-LX) of this antigen. The growth rate of these cells are similar. A mucin-like glycoprotein reactive with monoclonal antibody FH6 was detected after electrophoretic separation of KM12-HX cell lysates but not of KM12-LX lysates. The degree of invasion was compared in assays in vitro using matrigel-coated filter membranes. The number of KM12-HX cells that invaded the membranes was significantly higher than KM12-LX cells.

Topics: Antibodies, Monoclonal; Cell Division; Cell Separation; Colorectal Neoplasms; Genetic Variation; Glycoproteins; Humans; Lewis X Antigen; Molecular Weight; N-Acetylneuraminic Acid; Neoplasm Invasiveness; Neoplasm Metastasis; Sialic Acids; Tumor Cells, Cultured

Topics: Biomarkers, Tumor; Carcinoembryonic Antigen; Colorectal Neoplasms; Humans; Lewis X Antigen

Sialylated carbohydrate antigens, such as CA19-9 (sialyl Lea), CA-50 (sialyl Le4), CSLEX1 (sialyl Lex) and SLX (sialyl Lex-i), were assayed in the same preoperative serum samples of 63 patients with colorectal cancer, and compared with CEA. In addition immunohistochemical expressions of sialyl Lea, sialyl lex and sialyl Lex-i antigens were studied in 62 colorectal carcinomas and 42 normal mucosal sites remote from the malignant lesion using monoclonal antibodies CSLEA1, CSLEX1 and FH-6, respectively, in order to elucidate their tumor-specificity and clinical usefulness as a tumor-associated antigen. Serologically, the percent positive rates of CA19-9, CA-50, CSLEX1, SLX and CEA were 30.2%, 17.7%, 23.8%, 16.1% and 44.4%, respectively. In dukes' A and B, these sialylated carbohydrate antigens, especially CSLEX1 and SLX, showed low positive rates, but the percent positive rates of CSLEX1 and SLX correlated with operative radicality. The positive spectrum of CSLEX1 differed from that of CA19-9 in sera, and CEA had no correlation with these two antigens. The immunohistochemical expression rates of sialyl Lea, sialyl Lex and sialyl Lex-i were 88.1%, 17.0% and 9.5% in normal mucosa, but were 77.8%, 90.5% and 71.4% in carcinoma, respectively. These data suggested that the type 2 chain antigens CSLEX1 and SLX, which have high tumor-specificity compared with CA19-9, may be useful in preoperative diagnosis for extension of carcinoma and operative radicality, although early diagnosis using these sialylated carbohydrate antigens may be difficult, while the combined use of CA19-9, CSLEX1 and CEA should make it possible to detect a wide range of colorectal cancer patients.

Topics: Antibodies, Monoclonal; Antibodies, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Colorectal Neoplasms; Epitopes; Humans; Immunoenzyme Techniques; Immunohistochemistry; Lewis X Antigen

Sialyl Lewis(x)-i (SLX) as a new tumor-associated antigen was assayed in sera of 133 gastric cancer and 62 colorectal cancer patients with preoperative state in order to elucidate the clinico-pathological features. The percent positives in sera of patients with gastric and colorectal cancer were 12.7% (17/133) and 16.1% (10/62), respectively, and there were few positive cases in the early stages. Therefore, it is suggested that the early diagnosis for gastric and colorectal cancer using SLX alone may be difficult. However, it is revealed that SLX has a high tumor-specificity in consideration of the clinico-pathological properties such as H, P, n, ly, v factors, depth of cancer invasion and operative radicality. These data suggested that SLX could be applicable as a useful tumor-marker which shows the metastasis or extension of carcinoma.

Topics: Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Colorectal Neoplasms; Glycolipids; Humans; Lewis X Antigen; Middle Aged; Neoplasm Invasiveness; Neoplasm Metastasis; Neoplasm Staging; Predictive Value of Tests; Stomach Neoplasms

The Y haptenic blood group determinant is expressed on the cells of malignant gastrointestinal tumours and on the normal gastrointestinal epithelium of individuals who can secrete blood group antigens. an enzyme-linked immunosorbent assay incorporating the high affinity biotin-avidin interaction was developed to measure the serum levels of antigens bearing the Y hapten in patients with colorectal cancer. The specificity of the assays was between 88 and 93% and the sensitivity in detecting extensive disease was between 24 and 34%. However, up to 67% of patients with local or abdominal recurrent disease secreted antigens expressing the Y hapten, whereas only 30% of patients with overt hepatic metastases secreted a similar antigen. Recognition of antigens bearing the Y hapten may therefore be useful in detecting early local relapse of colorectal cancer when a second operation to excise the recurrence may be possible.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Antigens, Surface; Colorectal Neoplasms; Enzyme-Linked Immunosorbent Assay; Glycolipids; Humans; Lewis X Antigen; Methods; Neoplasm Recurrence, Local

We collected a total of 78 tissue specimens, including primary colorectal carcinoma, normal colonic mucosa, and liver metastases of colon carcinoma, to examine whether the extracts of these tissues inhibited the binding of a monoclonal antibody FH6, specific for sialyl-dimeric LeX antigen. The results of inhibition assays demonstrated that: (a) contents of FH6-reactive molecules were greater in carcinoma tissues than in normal colonic mucosa; (b) metastatic foci in livers contained more FH6-reactive molecules than primary tumors; (c) primary tumors from Dukes' stage B1 patients contained less FH6-reactive molecules than primary tumors from Dukes' stage D patients. The inhibitory activity of these tumor tissue extracts against the binding of a monoclonal antibody FH6 to cultured colon carcinoma cells was eliminated by prior treatment of the extracts with sialidase, confirming that the FH6-reactive materials were sialyl-dimeric LeX antigen. Electrophoretic separation of tumor tissue extracts on 3% polyacrylamide gels followed by direct staining with monoclonal antibody FH6 revealed that very high molecular weight glycoproteins, presumably mucins, contained sialyl-dimeric LeX antigen.

Topics: Antibodies, Monoclonal; Antigens, Neoplasm; Binding, Competitive; Colorectal Neoplasms; Electrophoresis, Polyacrylamide Gel; Glycolipids; Humans; Indicators and Reagents; Intestinal Mucosa; Lewis X Antigen; Liver Neoplasms; Neuraminidase

This study was undertaken to determine the value of SLX and CA19-9 in the tissue and serum in colorectal cancer and adenoma including the relation to Lewis blood type. SLX isolated by Hakomori et al is one of sialyl SSEA-1 antigens characterized by type 2 chain lacto series glycolipids. Distribution of SLX and CA19-9 was determined in 137 specimens of colorectal cancer (107 advanced and 30 early cancer) and 20 specimens of adenoma by using immunoperoxidase (ABC) method. Serum SLX, CA19-9 and CEA levels were measured preoperatively in 58 cases with histologically proven colorectal cancer by using RIA kits. Simultaneously, Lewis blood type were determined by hemagglutination test in 31 cases. The following results were obtained; 1) SLX and CA19-9 were mostly observed in apical membrane and luminal contents, but a few in cytoplasma of cancer tissue. Positive rate of SLX and CA19-9 was 96.2% and 88.8%, respectively. 2) SLX and CA19-9 were lightly stained in the goblet cells and surface epithelium in some of the mucosa adjacent to the cancer tissue, and positive rate of SLX and CA19-9 was 24.3% and 32.7%, respectively. 3) Positive rate of SLX and CA19-9 was 60% and 50% in adenoma, and 94% and 80% in "m" cancer. Positive rate was not different due to grade of dysplasia of adenoma, but distribution of SLX or grade of the staining of CA19-9 was different between adenoma with mild and moderate dysplasia.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Adenocarcinoma; Adenoma; Antigens, Neoplasm; Antigens, Tumor-Associated, Carbohydrate; Biomarkers, Tumor; Colorectal Neoplasms; Glycolipids; Humans; Immunohistochemistry; Lewis Blood Group Antigens; Lewis X Antigen