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

Nuclear texture reflects the overall structures of the chromatin organization. We recently reported the principles and prognostic importance of image analysis of nuclei from metastatic prostate cancer. Immunohistochemical up regulation of the adhesion molecule sialyl Lewis(x) is also reported to be a prognostic parameter. Presently we analyzed statistically the prognostic impact of these 2 new parameters compared to well-known clinical parameters in metastatic prostate cancer.. Prognostic factors, such as sedimentation rate, alkaline and acid phosphatases, hemoglobin, testosterone, performance status, pain due to metastasis, T category, histological grade and patient age, were included in a multivariate Cox proportional hazards regression analysis based on 262 patients from the Scandinavian Prostatic Cancer Group Study-2. Extent of bone lesions, deoxyribonucleic acid ploidy, texture analysis and sialyl Lewis(x) molecules based on subsets of these 262 patients were also analyzed in the same multivariate model.. This test identified chromatin texture as the most important factor (p < 0.001), followed by reaction of the oligosaccharide sialyl Lewis(x) (p < 0.01). Among the routine clinical and laboratory data, sedimentation rate, alkaline phosphatase and hemoglobin (p < 0.05) showed prognostic importance. Performance status, pain due to metastasis and extent of bone lesions showed prognostic value in the univariate analysis (p < 0.05).. These data indicate that computerized nuclear texture analysis as well as up regulation of sialyl Lewis(x) molecules may be new important prognostic factors in metastatic prostate cancer. Furthermore the prognostic importance of sedimentation rate, alkaline phosphatase and hemoglobin was confirmed.

Topics: Aged; Humans; Lewis X Antigen; Male; Middle Aged; Multivariate Analysis; Neoplasm Metastasis; Neoplasm Staging; Oligosaccharides; Prognosis; Proportional Hazards Models; Prospective Studies; Prostatic Neoplasms; Retrospective Studies; Sialyl Lewis X Antigen; Survival Rate

Protein glycosylation is a common posttranslational modification and glycan structural changes have been observed in several malignancies including prostate cancer. We hypothesized that altered glycosylation could be related to differences in gene expression levels of glycoprotein synthetic enzymes between normal and malignant prostate tissues.. We interrogated prostate cancer gene expression data for reproducible changes in expression of glycoprotein synthetic enzymes. Over-expression of GCNT1 was validated in prostate samples using RT-PCR. ELISA was used to measure core 2 O-linked glycan sialyl Lewis X (sLe(x) ) of prostate specific antigen (PSA), Mucin1 (MUC1), and prostatic acidic phosphatase (PAP) proteins.. A key glycosyltransferase, GCNT1, was consistently over-expressed in several prostate cancer gene expression datasets. RT-PCR confirmed increased transcript levels in cancer samples compared to normal prostate tissue in fresh-frozen prostate tissue samples. ELISA using PSA, PAP, and MUC1 capture antibodies and a specific core 2 O-linked sLe(x) detection antibody demonstrated elevation of this glycan structure in cancer compared to normal tissues for MUC1 (P = 0.01), PSA (P = 0.03) and near significant differences in PAP sLe(x) levels (P = 0.06). MUC1, PSA and PAP protein levels alone were not significantly different between paired normal and malignant prostate samples.. GCNT1 is over-expressed in prostate cancer and is associated with higher levels of core 2 O-sLe(x) in PSA, PAP and MUC1 proteins. Alterations of O-linked glycosylation could be important in prostate cancer biology and could provide a new avenue for development of prostate cancer specific glycoprotein biomarkers.

Topics: Acid Phosphatase; Aged; Glycosylation; Humans; Lewis X Antigen; Male; Middle Aged; Mucin-1; N-Acetylglucosaminyltransferases; Prostate-Specific Antigen; Prostatic Neoplasms; Protein Tyrosine Phosphatases; Sialyl Lewis X Antigen

Sialyl Lewis x (sLe(x)) plays an important role in cancer metastasis. But, the mechanism for its production in metastatic cancers remains unclear. The objective of current study was to examine the effects of a proinflammatory cytokine on the expression of glycosyltransferase and sulfotransferase genes involved in the synthesis of selectin ligands in a prostate cancer cell line. Androgen-independent human lymph node-derived metastatic prostate cancer cells (C-81 LNCaP), which express functional androgen receptor and mimic the castration-resistant advanced prostate cancer, were used. TNFα treatment of these cells increased their binding to P-, E- and L-selectins, anti-sLe(x) antibody, and anti-6-sulfo-sialyl Lewis x antibody by 12%, 240%, 43%, 248% and 21%, respectively. Also, the expression of C2GnT-1, B4GalT1, GlcNAc6ST3, and ST3Gal3 genes was significantly upregulated. Further treatment of TNFα-treated cells with either anti-sLe(x) antibody or E-selectin significantly suppressed their in vitro migration (81% and 52%, respectively) and invasion (45% and 56%, respectively). Our data indicate that TNFα treatment enhances the motility and invasion properties of LNCaP C-81 cells by increasing the formation of selectin ligands through stimulation of the expression of selective glycosyl- and sulfotransferase genes. These results support the hypothesis that inflammation contributes to cancer metastasis.

Topics: Cell Line, Tumor; Cell Movement; E-Selectin; Gene Expression Regulation, Enzymologic; Gene Expression Regulation, Neoplastic; Glycosyltransferases; Humans; L-Selectin; Ligands; Male; Neoplasm Invasiveness; Oligosaccharides; P-Selectin; Prostatic Neoplasms; Sialyl Lewis X Antigen; Sulfotransferases; Tumor Necrosis Factor-alpha

How cancer cells bind to vascular surfaces and extravasate into target organs is an underappreciated, yet essential step in metastasis. We postulate that the metastatic process involves discrete adhesive interactions between circulating cancer cells and microvascular endothelial cells. Sialyl Lewis X (sLe(X)) on prostate cancer (PCa) cells is thought to promote metastasis by mediating PCa cell binding to microvascular endothelial (E)-selectin. Yet, regulation of sLe(X) and related E-selectin ligand expression in PCa cells is a poorly understood factor in PCa metastasis. Here, we describe a glycobiological mechanism regulating E-selectin-mediated adhesion and metastatic potential of PCa cells. We demonstrate that alpha1,3 fucosyltransferases (FT) 3, 6, and 7 are markedly elevated in bone- and liver-metastatic PCa and dictate synthesis of sLe(X) and E-selectin ligands on metastatic PCa cells. Upregulated FT3, FT6, or FT7 expression induced robust PCa PC-3 cell adhesion to bone marrow (BM) endothelium and to inflamed postcapillary venules in an E-selectin-dependent manner. Membrane proteins, CD44, carcinoembryonic antigen (CEA), podocalyxin-like protein (PCLP), and melanoma cell adhesion molecule (MCAM) were major scaffolds presenting E-selectin-binding determinants on FT-upregulated PC-3 cells. Furthermore, elevated FT7 expression promoted PC-3 cell trafficking to and retention in BM through an E-selectin dependent event. These results indicate that alpha1,3 FTs could enhance metastatic efficiency of PCa by triggering an E-selectin-dependent trafficking mechanism.

Topics: Bone Marrow; Bone Marrow Neoplasms; Carcinoembryonic Antigen; CD146 Antigen; Cell Adhesion; Cell Movement; E-Selectin; Fucosyltransferases; Humans; Hyaluronan Receptors; Liver; Liver Neoplasms; Male; Neoplasm Metastasis; Oligosaccharides; Prostatic Neoplasms; Sialoglycoproteins; Sialyl Lewis X Antigen

Prostate cancer (PCa) cell tethering and rolling on microvascular endothelium has been proposed to promote the extravasation of PCa cells. We have shown that these adhesive events are mediated through binding interactions between endothelial (E)-selectin and Lewis carbohydrates on PCa cells. Prior data indicate that E-selectin-mediated rolling of bone-metastatic PCa MDA PCa 2b (MDA) cells is dependent on sialyl Lewis X (sLe(X))-bearing glycoproteins. To explore the molecular basis of sLe(X) synthesis and E-selectin ligand (ESL) activity on PCa cells, we compared and contrasted the expression level of glycosyltransferases, characteristically involved in sLe(X) and ESL synthesis, in ESL(+) MDA cells among other ESL(-) metastatic PCa cell lines. We also created and examined ESL(hi) and ESL(lo) variants of MDA cells to provide a direct comparison of the glycosyltransferase expression level. We found that normal prostate tissue and all metastatic PCa cell lines expressed glycosyltransferases required for sialo-lactosamine synthesis, including N-acetylglucosaminyl-, galactosyl-, and sialyltransferases. However, compared with expression in normal prostate tissue, ESL(+) MDA cells expressed a 31- and 10-fold higher level of alpha1,3 fucosyltransferases (FT) 3 and 6, respectively. Moreover, FT3 and FT6 were expressed at 2- to 354-fold lower levels in ESL(-) PCa cell lines. Consistent with these findings, ESL(hi) MDA cells expressed a 131- and 51-fold higher level of FT3 and FT6, respectively, compared with expression in ESL(lo) MDA cells. We also noted that alpha1,3 FT7 was expressed at a 5-fold greater level in ESL(hi) MDA cells. Furthermore, ESL(lo) MDA cells did not display sLe(X) on glycoproteins capable of bearing sLe(X), notably P-selectin glycoprotein ligand-1. These results implicate the importance of alpha1,3 FT3, FT6, and/or FT7 in sLe(X) and ESL synthesis on metastatic PCa cells.

Topics: Cell Line, Tumor; E-Selectin; Endothelial Cells; Glycosyltransferases; Humans; Ligands; Male; Microcirculation; Neoplasm Metastasis; Oligosaccharides; Prostatic Neoplasms; Protein Binding; Sialyl Lewis X Antigen

Prostate tumor cells preferentially adhere to bone marrow endothelial cells (BMECs) compared with endothelial linings from other tissue microvessels, implicating the importance of BMEC adhesion in the predilection of prostate tumor metastasis to bone. E (endothelial)-selectin, which functions as an initiator of leukocyte adhesion to target tissue endothelium, is constitutively expressed on BMECs, suggesting that prostate tumor cells could use this adhesive mechanism to initiate their migration into bone. In this report, we demonstrate for the first time that human bone-metastatic prostate tumor cells roll on human BMECs under physiological flow conditions. We show that these dynamic adhesive interactions are dependent on the expression of BMEC E-selectin and sialylated glycoconjugates on bone-metastatic prostate tumor cells. We also establish the importance of both glycoprotein(s) and glycosphingolipid structures displaying sialyl Lewis X epitopes as potential E-selectin ligands on bone-metastatic prostate tumor cells. Coexpression of sialylated glycoproteins and glycolipids on bone-metastatic prostate tumor cells triggers robust E-selectin binding activity, which is identical to that observed on human hematopoietic progenitor cells. By Western blot analysis, we identify candidate E-selectin glycoprotein ligand(s); distinct sialyl Lewis X (or HECA-452 antigen)-bearing membrane proteins were resolved at M(r) 130,000 and M(r) 220,000 as well as others ranging from M(r) 100,000 to M(r) 220,000. Immunohistochemical analysis of HECA-452 antigen expression on normal prostate tissue and on low- and high-grade prostate adenocarcinoma shows that HECA-452 antigen expression is directly associated with prostate tumor progression and may indicate acquisition of E-selectin ligand expression. These findings provide novel insight into potential adhesive mechanisms promoting hematogenous dissemination of prostate tumor cells into bone.

Topics: Adenocarcinoma; Antigens, Tumor-Associated, Carbohydrate; Bone Marrow Cells; Bone Neoplasms; Cell Adhesion; Cells, Cultured; Disease Progression; E-Selectin; Endothelium, Vascular; Glycoconjugates; Glycoproteins; Glycosphingolipids; Hematopoietic Stem Cells; Humans; Lewis X Antigen; Ligands; Male; Membrane Proteins; Oligosaccharides; Prostate; Prostatic Neoplasms; Sialyl Lewis X Antigen

Elevated expression of sialyl Lewis X has been postulated to be a prognostic indicator of prostate cancer. However, direct evidence for the relationship between increased expression of sialyl Lewis X and malignancy of prostate cancer is still lacking. To determine whether increased levels of sialyl Lewis X leads to malignancy in prostate tumor, we transfected the human prostate cancer cell line PC-3 with alpha1,3-fucosyltransferase III (FTIII) to obtain stable transfectants, PC-3-FTIII lines, that highly express sialyl Lewis X. When inoculated in the prostate of nude mice, PC-3-FTIII cells produced large prostate tumors, while mock-transfected PC-3 cells, which are negative for sialyl Lewis X antigen, produced small prostate tumors. The aggressive tumor formation by PC-3-FTIII cells was inhibited by preincubation of the tumor cells with anti-sialyl Lewis X antibody, by the presence of sialyl Lewis X oligosaccharide or by selectin ligand mimic peptide but not by control peptide. PC-3-FTIII cells and mock-transfected PC-3 cells exhibited no significant difference in cell numbers when cultured in vitro. Remarkably, PC-3-FTIII adhered to prostatic stromal cells in vitro with higher affinity than mock-transfected PC-3. Such adhesion was inhibited by preincubation of PC-3-FTIII cells with antisialyl Lewis X antibody, by the addition of sialyl Lewis X oligosaccharide or by selectin ligand mimic peptide. However, anti-E-selectin, anti-P-selectin or anti-L-selectin antibodies did not inhibit the adhesion of PC-3-FTIII cells to the stromal cells. These results suggest that prostate cancer cells gain aggressiveness through adhesive interaction with prostatic stromal cells by a novel mechanism involving sialyl Lewis X.

Topics: Adenocarcinoma; Animals; Biopsy; Cell Adhesion; Cell Division; Cell Movement; Fucosyltransferases; Gene Transfer Techniques; Humans; Immunohistochemistry; Male; Mice; Mice, Nude; Neoplasm Invasiveness; Neoplasm Staging; Oligosaccharides; Plasmids; Prostatic Neoplasms; Recombinant Proteins; Sialyl Lewis X Antigen; Stromal Cells; Transplantation, Heterologous; Tumor Cells, Cultured

Recently, it has been reported that upregulation of the oligosaccharide sialyl Le(x) (SLe[x]) in prostate cancer is associated with hormone-resistant, aggressive disease. However, it is not clear that SLe(x) expressed on prostate cancer cells has a biological function related to metastatic potential.. The expression levels of SLe(x), sialyl Le(a) (SLe[a]), disialosyl galactosylgloboside (DSGG), monosialosyl galactosylgloboside (MSGG) and variousfucosyltransferases in 3 prostate cancer cell lines were determined. The function of SLe(x) expressed on prostate cancer cell lines was determined by a selectin-dependent adhesion assay.. No prostate cancer cell lines expressed SLe(a), DSGG, or MSGG, but all prostate cancer cells moderately expressed SLe(x). Fucosyltransferase expression did not correlate with the expression of SLe(x), and all prostate cancer cells failed to bind immobilized selectin.. The expression of SLe(x) on these prostate cancer cells does not correlate with selectin-dependent adhesion.

Topics: Cell Adhesion; Cell Line; Endothelium, Vascular; Flow Cytometry; Fucosyltransferases; Humans; Male; Oligosaccharides; Prostatic Neoplasms; Selectins; Sialyl Lewis X Antigen

Early stage prostate cancers are now commonly encountered because of widespread use of screening tools. Increased cancer cell proliferation, and expression of sialyl Lewis could be important as predictors of clinical behaviour and survival in addition to histologic grade. In this study, the expression of sialyl-LewisX (sLeX) was determined by immunohistochemical methods in 38 routinely processed prostate biopsies and transurethral resections preceding radical prostatectomies for organ confined prostate cancers. Histologic grades were determined from pathologic reports and divided into two (2) groups; low grade (Gleason score 2-4) and medium grade (Gleason score 5-7). Tumour stages were based on radical prostatectomy reports and 29 were T2 and 9 were T3. SLeX was positive in 10 of 14 (71.4%) low grade and 14 of 24 (62.5%) medium grade cancers; 22/29 (75.9%) T2 and 8/9 (88.9%) T3 were sLeX positive; 1 of 15 (7.2%) low grade and 5 of 24 (20.8%) medium grade were strongly positive (3+) or overexpressing sLeX. Overexpression of sLeX was a feature of medium grade cancer, suggesting that localized prostate cancers with increased potential for progression and metastasis exist in the clinically non-metastatic group.

Topics: Biopsy; Humans; Immunohistochemistry; Male; Neoplasm Staging; Oligosaccharides; Prostatic Neoplasms; Sialyl Lewis X Antigen

Alteration of cell surface carbohydrate antigens during malignant transformation is a well-known phenomenon observed in various tumors. In prostatic carcinoma, nearly total deletion of normally occurring ABO and type I-based Lewis antigens, Le(a) and Leb, has been observed in several studies. We studied expression of the closely related type II antigens Le(x), Le(y), and sialyl-Lewis(x) (SLe(x)) using monoclonal antibodies. Thirty formalin-fixed specimens obtained from radical prostatectomy, containing prostatic carcinoma as well as benign tissue, were evaluated by immunohistochemistry. In both cancer and benign tissue, Le(x) expression was minimal or absent. In benign tissue, Le(y) was expressed in ducts and in the basal layer of glandular epithelium. In tumor tissue, Le(y) expression was greatly increased and extensive staining was observed in 26 of 30 cases. The SLe(x) expression in benign tissue was observed only in larger ducts, never in glandular secretory epithelial cells. In carcinoma, rare cells positive for SLe(x) were present in 8 of 30 cases, and stronger expression with focal to patchy distribution was observed in 14 of 30 cases. The results suggest an alteration in glycosyl transferase activity in prostatic carcinoma, with preserved or increased activity of enzymes responsible for the synthesis of the type II core sequence. This sequence is further glycosylated and expressed as the difucosylated compound Le(y) or the monofucosyl, monosialyl compound SLe(x). For prostate, Le(y) and SLe(x) are the only blood group-related antigens known to be minimal or absent in benign secretory epithelial cells that are more highly expressed in malignant tissue.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Biomarkers, Tumor; Humans; Lewis Blood Group Antigens; Lewis X Antigen; Male; Oligosaccharides; Prostatic Neoplasms; Sialyl Lewis X Antigen

Metastatic prostate cancer has an unpredictable long-term prognosis. At present, there are few specific predictors to indicate the outcome for the individual patient. We have studied immunoreactivity for type-2 carbohydrate structures, known to be involved in various cell adhesion processes, in patients with metastatic prostate cancer. One group of patients (n = 26) did not progress within 3 years after orchiectomy, while another group of patients (n = 33) progressed within 1 year following castration and survived less than 2 years. Among the parameters studied, sialyl LewisX carbohydrate up-regulation was the only variable showing significant association with poor prognosis (P < 0.01). Sialyl LewisX discriminated between these two outcome groups with 71% predictability and 96% specificity. Our results indicate that up-regulation of sialyl LewisX is associated with hormonal-resistant, aggressive disease. This prognostic marker may, therefore, have an important role in selecting proper treatment for patients with metastatic prostate cancer.

Topics: Adenocarcinoma; Aged; Aged, 80 and over; Biomarkers, Tumor; Carbohydrate Sequence; Humans; Immunohistochemistry; Lewis X Antigen; Male; Middle Aged; Molecular Sequence Data; Neoplasm Metastasis; Oligosaccharides; Predictive Value of Tests; Prognosis; Prostatic Neoplasms; Sialyl Lewis X Antigen; Up-Regulation