lewis-y-antigen and Carcinoma--Squamous-Cell

Several mechanisms are involved in the development of resistance to therapy in locally advanced cervical squamous cell carcinoma (LACSCC). Studies have shown that CD44 and Lewis Y antigen (LeY) form a complex that is associated with chemoresistance, tumor invasion and metastasis. We assessed the role of CD44 and LeY in the outcome of LACSCC patients treated with different chemotherapy regimens.. 126 LACSCC patients at FIGO stages IIB-IVA were selected from the GOCS database: 74 patients included in 3 different prospective phase II trials in the neoadjuvant setting (vinorelbine, docetaxel, ifosfamide-vinorelbine-cisplatin) and 52 patients treated with standard radiochemotherapy based on cisplatin (RCBC). Clinical data at baseline, disease-free survival (DFS) and overall survival (OS) were recorded. Univariate and multivariate Cox models were employed.. Median age was 45.6 years (range: 24.9-80.5). Sixty-three and 47 tumors were CD44+ and LeY+, respectively. Tumors with expansive growth showed higher grade (p = 0.0024), mitotic index (p = 0.0505), tumor necrosis (p = 0.0191), LeY+ (p = 0.0034) and CD44+/LeY+ coexpression (p = 0.0334). CD44+ cells were present in 91.3% of patients with local recurrence (p = 0.0317). Advanced stage was associated with LeY+ tumors. Patients treated with RCBC had worse DFS and OS when their tumors expressed LeY (p = 0.0083 and p = 0.0137, respectively). Pre-treatment hemoglobin level, FIGO stage and tumor response remained the most significant prognostic factors in Cox regression.. In our cohort of LACSCC patients, the coexpression of CD44 and LeY was not associated with worse outcome. However, in the subgroup of patients receiving RCBC, LeY expression was correlated with shorter DFS and OS.

Topics: Adult; Antineoplastic Agents; Carcinoma, Squamous Cell; Cisplatin; Female; Humans; Lewis Blood Group Antigens; Neoplasm Staging; Uterine Cervical Neoplasms

Aberrant glycosylation changes normal cellular functions and represents a specific hallmark of cancer. Lewisy (Ley) carbohydrate upregulation has been reported in a variety of cancers, including oral squamous cell carcinoma (OSCC). A high level of Ley expression is related to poor prognosis of patients with oral cancer. However, it is unclear how Ley mediates oral cancer progression. In this study, the role of Ley in OSCC was explored. Our data showed that Ley was upregulated in HSC-3 and OC-2 OSCC cell lines. Particularly, glycosylation of epidermal growth factor receptor (EGFR) with Ley was found in OC-2 cells, and this modification was absent upon inhibition of Ley synthesis. The absence of Ley glycosylation of EGFR weakened phosphorylation of AKT and ERK in response to epidermal growth factor (EGF). Additionally, EGF-triggered cell migration was reduced, but cell proliferation was not affected. Ley modification stabilized EGFR upon ligand activation. Conversely, absence of Ley glycosylation accelerated EGFR degradation. In summary, these results indicate that increased expression of Ley in OSCC cells is able to promote cell migration by modifying EGFR which in turn stabilizes EGFR expression and downstream signaling. Targeting Ley on EGFR could have a potential therapeutic effect on oral cancer.

Topics: Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Movement; Cells, Cultured; ErbB Receptors; Extracellular Signal-Regulated MAP Kinases; Glycosylation; Humans; Keratinocytes; Lewis Blood Group Antigens; Male; Mouth Neoplasms; Protein Processing, Post-Translational; Proto-Oncogene Proteins c-akt

Expression and implication of carbohydrate antigens in squamous cell carcinomas (SCCs) in oral cavity was examined. In the cell lines, type 2H and Lewis y antigens were markedly expressed. In the tissues from SCC patients and benign disorders, type 2H was highly expressed in hyperplasia (96.4 %), displasia (92.9 %) and SCC (100 %). Lewis y was, in turn, expressed mainly in cancer tissues (91.3 %), suggesting that Lewis y is a cancer-associated antigen. Normal oral mucosa showed no expression of these blood group antigens. Surprisingly, Lewis y antigen disappeared in the invasion sites where Ki-67 was definitely stained. Over-expression of Lewis y with manipulation of a fucosyltransferase cDNA resulted in suppression of cell growth and invasion, and knockdown of Lewis y also brought about increased cell growth and invasion. In either situations, no changes in the expression of sialyl-Lewis x could be found. Lowered tumor growth and invasion into surrounding tissues were also shown in Lewis y-positive SCC grafts in nu/nu mice. All these results together with alternative staining between Lewis y and Ki-67 in cancer tissues and FUT1 transfectants suggested that loss of Lewis y is a crucial event for the late stage of SCCs.

Topics: Animals; Carcinoma, Squamous Cell; Cell Line, Tumor; Fucosyltransferases; Galactoside 2-alpha-L-fucosyltransferase; Gene Expression Regulation, Neoplastic; Humans; Ki-67 Antigen; Lewis Blood Group Antigens; Mice; Mice, Inbred BALB C; Mouth Mucosa; Mouth Neoplasms; Neoplasm Invasiveness; Neoplasm Transplantation; Oligosaccharides; Organ Specificity; Sialyl Lewis X Antigen

B3 antibody specifically binds the LewisY-related carbohydrate antigen of many carcinomas, and it is used as a model antibody in this study. In a previous study, the Fab fragment of the antibody was fused to a 38 kDa truncated form of Pseudomonas exotoxin A, PE38, to make Fab- PE38, where PE38 is fused to the Fd fragment of the Fab domain. This parent monomer molecule, Fab-PE38, had no cysteine in the hinge region, and it could not make a disulfide bond to form a disulfide bond bridged homodimer. In this study, we constructed three different kinds of divalent Fab-toxin fusion homodimers where the toxin is fused to the light chain of Fab, (Fab-PE38fl)2. In addition to the PE38 toxin fused to the light chain, these three molecules have different hinge sequences h1, h2, and h3 making Fabh1-, Fabh2-, and Fabh3-PE38fl monomers, respectively. These hinges contain only one cysteine on different positions of the hinge sequence. The disulfide bond between the hinge region of two monomers forms homodimers (Fabh1-PE38fl)2, (Fabh2-PE38fl)2, and (Fabh3- PE38fl)2. The refolding yields of these dimers were 5- 16-fold higher than a previously constructed dimer where the PE38 was fused to the Fd fragment (Fabh1-PE38)2. Our data suggest that the steric repulsion between the two PE38s in (Fabh1-PE38)2 during disulfide bridge formation is relieved by fusing it at the end of the light chain. The best cytotoxicity value of these dimers showed about 2.5-fold higher on an MCF7 cell line than that of the monovalent reference molecule in ng/ml scale, which is 15-fold higher in pM scale.

Topics: Antibodies, Monoclonal; Bacterial Proteins; Biotechnology; Breast Neoplasms; Carcinoma, Squamous Cell; Cell Line, Tumor; Cell Survival; Disulfides; Escherichia coli; Humans; Immunoglobulin Fab Fragments; Immunoglobulin Light Chains; Immunotoxins; Lewis Blood Group Antigens; Plasmids; Protein Folding; Recombinant Fusion Proteins

The majority of cancer cells derived from epithelial tissue express Lewis-Y (LeY) type difucosylated oligosaccharides on their plasma membrane. This results in the modification of cell surface receptors by the LeY antigen. We used the epidermal growth factor (EGF) receptor family members ErbB1 and ErbB2 as model systems to investigate whether the sugar moiety can be exploited to block signaling by growth factor receptors in human tumor cells (i.e., SKBR-3 and A431, derived from a breast cancer and a vulval carcinoma, respectively). The monoclonal anti-LeY antibody ABL364 and its humanized version IGN311 immunoprecipitated ErbB1 and ErbB2 from detergent lysates of A431 and SKBR-3, respectively. ABL364 and IGN311 blocked EGF- and heregulin-stimulated phosphorylation of mitogen-activated protein kinase [MAPK = extracellular signal-regulated kinase 1/2] in SKBR-3 and A431 cells. The effect was comparable in magnitude with that of trastuzumab (Herceptin) and apparently noncompetitive with respect to EGF. Stimulation of MAPK by ErbB was dynamin dependent and contingent on receptor internalization. ABL364 and IGN311 changed the intracellular localization of fluorescent EGF-containing endosomes and accelerated recycling of intracellular [(125)I]EGF to the plasma membrane. Taken together, these observations show that antibodies directed against carbohydrate side chains of ErbB receptors are capable of inhibiting ErbB-mediated signaling. The ability of these antibodies to reroute receptor trafficking provides a mechanistic explanation for their inhibitory action.

Topics: Antibodies, Monoclonal; Breast Neoplasms; Carcinoma, Squamous Cell; Cell Line, Tumor; Epidermal Growth Factor; ErbB Receptors; Female; Humans; Iodine Radioisotopes; Kinetics; Lewis Blood Group Antigens; MAP Kinase Signaling System; Mitogen-Activated Protein Kinases; Phosphorylation; Precipitin Tests; Receptor, ErbB-2; Tunicamycin; Vulvar Neoplasms

Tumor procoagulant is associated with cancer at advanced stages of malignancy such as infiltration and metastasis. In the present study, we investigated the role of Ley glycolipid in the mechanism of cancer metastasis. Ley glycolipid acts as an important cofactor in the expression of the blood-coagulating activity of cancer cell-derived coagulating activity 1 (CCA-1), which is one of the known tumor procoagulants. Monoclonal antibody (MoAb) FS01, which serves as the Ley-recognizing epitope, inhibits the procoagulant activity of CCA-1 was found to dose-dependently inhibit the procoagulant activity of normal plasma induced by the human lung adenocarcinoma cell line, HAL8, which shows a high level of Ley expression. It did not, however, inhibit the procoagulant activity of the human colon cancer cell line, RPMI4788, which does not express Ley. Administration of FS01 MoAb inhibited lung metastasis of HAL8 cells, but not that of RPMI4788. The absence of antibody-dependent cellular cytotoxicity and complement-mediated cytotoxicity of FS01 MoAb against the HAL8 cell line suggests that the inhibition of HAL8 metastasis by FS01 MoAb derives from the inhibition of blood-coagulating activity of the latter. These findings indicate that Ley glycolipid plays an important role in the mechanism of cancer metastasis via the procoagulant activity of CCA-1.

Topics: Adenocarcinoma; Animals; Antibodies, Monoclonal; Blood Coagulation Factors; Blood Coagulation Tests; Carcinoma, Squamous Cell; Colorectal Neoplasms; Cysteine Endopeptidases; Drug Screening Assays, Antitumor; Flow Cytometry; Glycolipids; Lewis Blood Group Antigens; Lung Neoplasms; Mice; Mice, Nude; Neoplasm Proteins; Tumor Cells, Cultured

The progression from uncontrolled cell proliferation to invasion and metastasis of epithelial tumors is partially understood. Alteration of epithelial mucin expression have been described in different malignant localizations but only few attempts have been made to identify mucin expression in malignant laryngeal tumors. In the present report, results are shown of studies on the expression of mucins and carbohydrate related antigens in laryngeal cancer and on the isolation of MUC1 mucin from this tumor tissue. Malignant laryngeal specimens were processed for immunohistochemical analysis and for extranuclear membrane fractions (ENM) which were obtained by ultracentrifugation. Subsequently, ENM samples were centrifuged in density-gradient; the analysis of fractions was performed by means of SDS-PAGE and Western-blotting. The panel of monoclonal antibodies (MAbs) included anti MUC1 mucin, anti Lewis x, anti sialyl Lewis x, anti Lewis y, anti MUC-5B, anti oral mucin (gp230), anti Tn hapten, anti p53 and anti cytokeratins. By immunohistochemistry, it was possible to detect MUC1 mucin, Lewis x and Lewis y showing strong reactions while sialyl-Lewis x and Tn antigen only reacted weakly in a few cells; cytokeratins were detected in all samples. In ENM derived fractions obtained by CsCl centrifugation, MUC1 was demonstrated by Western blotting.. (1) laryngeal cancer antigenic expression comprises mostly MUC1 mucin, Lewis x, Lewis y as well as Tn antigen and (2) the methodology here employed is useful to isolate MUC1 from tumor samples.

Topics: Aged; Antigens, Tumor-Associated, Carbohydrate; Blotting, Western; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; Laryngeal Neoplasms; Lewis Blood Group Antigens; Lewis X Antigen; Male; Mucin-1; Mucins

To investigate the prognostic significance of the carbohydrate epitopes H and Le(y) and their relationship with proliferation and apoptosis.. Eighty randomly selected patients with T1-T4 oral tongue squamous cell carcinoma (SCC) were studied. Serial sections were cut from diagnostic, formalin-fixed, paraffin-embedded specimens.. Sections were stained immunohistochemically for H antigen and Le(y).. Expression of H antigen was associated positively with Le(y) expression (P = .0001). Expressions of H antigen or Le(y) correlated with the proliferative markers Ki67 (P = .0442 and P = .0003, respectively) and pAgNOR > 1 (P = .0674 and P = .0047, respectively), but not with apoptotic markers such as Bax expression or the apoptotic index (AI). Tumors that expressed H antigen and high levels of Le(y) (> 50%) had a poor prognosis (P = .0006 and P = .0056, respectively). Combinations of expression of H antigen and Le(y), and either proliferative or apoptotic markers revealed an enhanced prognostic potential (P < .0001). The combination of pAgNOR score greater than 1 and H-antigen expression appeared to be the best combination to predict good prognosis.. The expression of H antigen and Le(y), especially their combination with proliferative or apoptotic markers, has prognostic value in tongue SCC.

Topics: ABO Blood-Group System; Apoptosis; Biomarkers, Tumor; Carcinoma, Squamous Cell; Female; Humans; Lewis Blood Group Antigens; Male; Middle Aged; Prognosis; Random Allocation; Tongue Neoplasms

We semiquantitatively analyzed expression of PCNA and LeY in seborrheic keratosis (SK), actinic keratosis (AK), Bowen's disease (BD), and squamous cell carcinoma (SCC), using immunocytochemically stained tissue sections. PCNA expression increased in a stepwise fashion from low levels in normal skin to higher expressions within SK, AK, BD, and SCC. The levels of LeY protein also increased in this order. The PCNA expression pattern shifted from expression limited to the basal and suprabasal cell layers (in normal skin and SK) to expression extending to the upper squamous and granular layers (in AK, BD, and SCC). On the other hand, the pattern of LeY expression shifted from the granular (in normal skin) to the upper squamous (in SK and AK) and suprabasal layers (in BD and SCC). These findings suggest that PCNA expression is related to the degree of cell proliferation and that LeY expression is related to the degree of differentiation or keratinization of tumor cells. In addition, PCNA and LeY show a reciprocal relationship in their expression.

Topics: Bowen's Disease; Carcinoma in Situ; Carcinoma, Squamous Cell; Humans; Immunohistochemistry; Keratosis, Seborrheic; Lewis Blood Group Antigens; Proliferating Cell Nuclear Antigen; Skin Neoplasms

To determine whether keratoacanthoma (KA) is unique or a variant of squamous cell carcinoma (SCC), the expressions of proliferating cell nuclear antigen (PCNA) and Le(Y) in 16 KA cases (11 fully developed and 5 involutional types; two of the involutional types transformed from the fully developed types) and 11 cases of well-differentiated SCC were investigated by immunohistochemistry. The monoclonal antibodies used were PC-10 for PCNA and BM-1/JIMRO for Le(Y). Le(Y) is one of the Lewis-type antigens, and is thought to be related to apoptosis. In most of the cases of fully developed KA, the PCNA expression was linear or band-like and was limited to the basal and suprabasal layers, while the Le(Y) expression was seen in a diffuse pattern throughout the epidermis, except for the basal layer. However, in 5 of these cases, there were some areas in the basal and suprabasal layer where PCNA-positive cells were few, while Le(Y) expression was very strong. In the involutional Ka cases, the PCNA expression was very weak in the basal and suprabasal layers, and the Le(Y) expression was strong throughout the epidermis. In the SCC cases, the PCNA and Le(Y) expressions were very strong and diffuse throughout the tumor masses except for the lack of PCNA in the horny layer and the lack of Le(Y) in the basal layer. These findings indicate that there are some differences between KA and SCC in the expression pattern of PCNA and Le(Y), and that apoptosis may occur in the Le(Y)-positive areas in the basal and suprabasal layers and thus cause the involution of KA.

Topics: Aged; Aged, 80 and over; Apoptosis; Carcinoma, Squamous Cell; Diagnosis, Differential; Female; Humans; Immunoenzyme Techniques; Keratoacanthoma; Lewis Blood Group Antigens; Male; Middle Aged; Proliferating Cell Nuclear Antigen; Skin Neoplasms