Lectin binding to prostatic adenocarcinoma

The binding of different lectins (concanavalin A [Con A], triticum vulgaris [WGA], glycine maximum [SBA], dolichos bilflorus [DBA], ulex europaeus [UEA I], arachis hypogaea [PNA], and ricinus communis [RCA I]) to cells of normal prostate glands, hyperplastic glands and adenocarcinoma was studied. The Con A, WGA, DBA, PNA and RCA I bound to both normal and hyperplastic glands. The binding in the malignant glands differed from that of the benign conditions. The SBA, which was not bound by benign cells, was bound to the malignant glandular cells. Also, UEA I was bound to a part of the carcinoma cells. In addition, the binding pattern of Con A and WGA in the cells differed between the malignant and benign conditions. Based on the results of this study, it is suggested that lectin histochemical study might be useful in routine pathologic examination to detect malignant cells in cases which are doubtful with regard to malignancy by routine methods.     

Lectin histochemistry of normal and neoplastic nasopharyngeal epithelium.

The cell surface carbohydrate profile of formalin-fixed paraffin-embedded tissue sections of normal and neoplastic epithelium was evaluated using 9 plant lectins. Three lectins, namely Con A, RCA and WGA, showed a similar pattern and staining intensity from normal epithelium to metaplastic squamous epithelium and nasopharyngeal intraepithelial neoplasia (NPIN). However, a decrease in staining reactivity was observed in undifferentiated nasopharyngeal carcinoma. Significant differences in intensity and distribution were seen in UEA and cryptic PNA residue (after neuraminidase pretreatment) from normal nasopharyngeal epithelium to NPIN. Infiltrative undifferentiated carcinomas showed a heterogenous lectin binding pattern and altered intensity of lectin binding in one case of DBA and three cases of PNA (no neuraminidase pretreatment), suggesting a variation in expression of carbohydrate by tumour cells. These results indicate that neoplasia in nasopharyngeal epithelium is associated with alterations in terminal sialic acid, -Fucose residues and -Gal-D-GalNac residues present in the outer parts of glycoconjugates. SBA, VVL and BSL failed to stain any types of epithelia. Desialylation of tissues by preincubation with neuraminidase did not expose DBA, SBA, VVL and BSL binding sites. These findings may be used as a baseline for evaluation of lectin binding in preinvasive and invasive lesions of the nasopharynx.     

Lectin binding sites of cultured ovarian Sertoli cell tumors and follicular granulosa cells

A panel of seven alkaline phosphatase labeled lectins was used to probe nitrocellulose electroblots of SDS-PAGE separated proteins from a primary culture of normal ovarian granulosa cells and an ENU-induced Sertoli cell tumor cell line (SCTL-I). Several additional lectin binding proteins were observed in silver stained SDS-PAGE gels as well as with lectins in SCTL-I. Succinated concanavalin A (Suc. Con A), Ricin communis agglutinin (RCA-I), Ulex europaeus agglutinin (UEA 1), Soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA) and Peanut agglutinin (PNA) stained more intensely in SCTL-I than normal granulosa cells. The same lectins as above, labeled with fluorescein isothiocyanate (FITC), were used to study the distribution of specific binding sites of tissue cultured cells grown in chamber/slides. Both normal ovarian granulosa cells and SCT cells exhibited strong peninuclear cytoplasmic labeling with Con A UEA-1 and WGA exhibited predominantly a nuclear and granular cytoplasmic staining pattern. SBA and DBA exhibited a strong coarse granular cytoplasmic labeling in granulosa cells and moderate granular cytoplasmic in SCT cells. In granulosa cells, Golgi regions stained strongly with PNA but weakly in SCT cells. RCA-I staining was negative in both cultures. Labeling of tissue cultured cells with lectins provides more details than histological sections of lectins binding sites at cellular structural levels.     

STUDY ON GLYCOCONJUGATE CHANGES ON CELL SURFACE IN PROGRESSIVE DEVELOPMENT OF PULMONARY TUMOR

Aim: To investigate glycoconjugate changes on the cell surface of proliferative lesions and neoplasms of mice lungs at various stages of tumorigenesis, the relation between progressive development of mouse pulmonary tumors and expression of cell surface saccharide. Materials and methods: Thirty - one male A/J strain mice at 5 weeks of age were treated intraperitoneally with a single injection of 20 - methyl-cholanthrene (20-MC), 292 pulmonary lesions including 31 hyperplasias, 145 alveolar adenomas, 61 papillary adenomas, 55 papillary adenocarcinomas and their combined type were obtained. The binding affinities of cells in normal respiratory epithelia and in proliferative lesions to four peroxidases - conjugated lectins, Madura pomifera agglutinin (MPA), Arachis hypogea agglutinin (PNA), Ricinus communis agglutinin (RCA), and wheat germ agglutinin (WGA) were examined. Results: Cells of hyperplasia and alveolar adenoma showed fairly strong affinity to all the four lectins. However, part of papillary adeno     

Brenner瘤细胞分化的凝集素受体标记

研究７种生物素化的凝集素对卵巢Ｂｒｅｎｎｅｒ瘤不同分化方向和分化性质瘤细胞的标记特点，麦胚素、花生素受体在瘤细胞呈顶浆型表达是瘤细胞向腺上皮分化的标志；双花扁豆素受体瘤细胞膜阳性显示其向鳞状上皮分化倾向；ＷＧＡ、ＰＮＡ、兀鹰素（ＢＳ－１）和刀豆素在瘤细胞浆内弥漫性结合增强表明Ｂｒｅｎｎｅｒ瘤由良性、增生性向恶性转化。凝集素受体表达差异可做为Ｂｒｅｎｎｅｒ瘤细胞的分化标志，体现该瘤上皮成分分化，失分     

Histochemical study of colorectal adenocarcinoma by fluorescein isothiocyanate (FITC) conjugated lectins

Binding of FITC conjugated lectins to mucin in benign and malignant colon lesions was studied by fluorescence microscopy. In 145 cases of colonic adenocarcinoma, PNA (peanut agglutinin) was found to be bound to 75% of cases, while DBA (dolichos biflorus agglutinin) to 13% only. By contrast, DBA was bound to 94% of normal colon mucosa, while PNA to 15% only. Chronic colitis, simple adenoma and inflammatory polyps had the same binding pattern as normal mucosa, but villous adenoma, mixed polyps and multiple polyposis which are considered as premalignant lesions in colon had high positive rate of PNA binding and low binding percentage of DBA. These results indicate that an exposed carbohydrate structure Gal B1-2----GlNAc is expressed in the mucin produced by colonic adenocarcinoma. Meanwhile, some normal mucin component disappears from the malignant colonic epithelium. The mechanism of alteration of human colonic mucin present in malignant transformation is briefly discussed.     

Lectin binding in murine tumour lines with different malignant characteristics. Effect of enzyme (pronase, neuraminidase) treatment on lectin labelling

We used 5 syngeneic murine tumour systems for studying quantitative lectin surface binding of intact viable tumour cells. We also investigated, for 3 of the tumours, whether the lectin binding sites were susceptible to proteolytic enzyme (pronase) or neuraminidase treatment. There were significant differences between two of the tumour lines in the binding of wheat germ agglutinin (WGA), concanavalin A (ConA). peanut agglutinin (PNA), soybean agglutinin (SBA) and Ulex europeus agglutinin (UEA). There were also variations in lectin binding between the other tumor lines, but these differences were not statistically significant. Lectin binding showed no evident relationship to the malignancy or the metastasis-forming capacity of the respective tumour cell line. Proteolytic treatment, which drastically affects intravenously induced experimental metastasis formation by one of the tumours, caused a decreased binding of SBA, ConA and WGA. Neuraminidase treatment increased both PNA and SBA binding in three different cell lines, presumably by removing sialic acid masking D-galactose and N-acetyl-D-galactose-amine groups.     

Lectin histochemistry of adenomatous polyps. Not a predictor of metachronous lesions

The binding characteristics of fluorescein isothiocyanate conjugated lectins to normal colonic mucosa, and 43 adenomatous polyps were studied by fluorescence microscopy. The lectin, Dolichos biflorus agglutinin (DBA) stained intensely to upper crypt cells of the sigmoid colon and rectum but to a lesser degree to proximal colonic crypts or lower crypt cells distally. Peanut agglutinin (PNA) and Ulex europaeus agglutinin (UEA) did not bind to the theca of proximal or distal colonic crypts. The lectin Griffonia simplicafolia agglutinin (GSA1) bound intensely to upper and lower crypt cells of both regions. PNA binding was noted in 56% of adenomatous polyps, occurred more often in polyps of the distal colorectum, and increased with polyp size and villous histology. UEA bound to 26% of adenomatous polyps, 42% of proximal polyps, and 17% of distal polyps. DBA staining was noted in 72% of polyps without regional preference. GSA1 stained all polyp specimens. To determine if the lectin binding characteristics of an index (initial) polyp might serve as a predictor of metachronous lesions, 20 patients (29 polyps) without a history of polyps or cancer and who had at least one surveillance colonoscopy 1 to 3 years after the initial polypectomy were studied. The presence or absence of PNA, UEA, or DBA binding in an index polyp did not predict the occurrence of metachronous lesions. Five of the six patients with more than one index polyp had metachronous polyps at follow-up surveillance colonoscopy.     

A STUDY OF LECTINS IN HUMAN BREAST CANCER AND PRECANCEROUS LESIONS

This paper report the binding pattern of 12 different kinds of lectin in human breast lesions. Of the 12 kinds of lectins, WGA showed the highest binding activity to the cells of the breast tissue derived; the binding of BSL, SBA and DBA were localized to membrane or cytoplasm of cancer cells and to the lumina membrane border of the normal and benign lesions; PNA receptor is related with the differentiation of the breast; cancer; we didn't find any relationship between the lectin receptor and the tendency of metastasis of breast cancer.     

Lectin binding in human breast cancer: clinical and pathologic correlations with fluorescein-conjugated peanut, wheat germ and concanavalin A binding

Cell surface glycoconjugates of human breast cancer tissue were investigated using FITC peanut (PNA), wheat germ (WGA) and jackbean (concanavalin A; Con A) agglutinins. Although PNA and WGA binding patterns differed when normal and malignant breast tissues were compared, the specificity of this finding was poor and neither PNA nor WGA binding correlated with oestrogen receptor status or clinical outcome. Con A binding on the other hand was not seen in normal breast tissue. The percentage of Con-A-positive (Con A+) tumours increased progressively with advancing stage (16% Con A+ in stage I, 40% Con A+ in stage III) and there was a trend towards longer disease-free survival of patients with Con A- as compared to those with Con A+ tumours. However, in a multivariate analysis it was found that these differences were explained by the association with the stage of the disease.     

Lectin binding glycoproteins in human melanoma cell lines with high or low tumorigenicity

Lectin binding glycoproteins of 5 human malignant melanoma cell lines (HMMCL), differing in their ability to grow subcutaneously in athymic nude mice, were compared by electrophoresis of total cellular proteins and subsequent incubation of SDS-poly-acrylamide gel with 125I-labelled lectins. Despite the similarity between the protein profiles of the different HMMCL, Concanavalia ensiformis agglutinin (ConA), wheat-germ agglutinin (WGA) and peanut agglutinin (PNA) revealed differences in their glycoprotein expression, in contrast with Ulex europaeus agglutinin I (UEA I). A great diversity was observed in the electrophoretic mobilities and/or staining intensities of ConA and WGA binding glycoproteins of HMMCL. However, neither ConA-reactive glycoproteins nor WGA-reactive glycoproteins could be detected that were characteristic of HMMCL with high tumorigenicity (HT) or low tumorigenicity (LT). In contrast, the expression of two cell-surface PNA binding glycoproteins appeared to be related to the tumorigenic phenotype of HMMCL. One of them, with an apparent molecular weight of 190 kDa, was only detected in the LT cell lines. The other, with an apparent molecular weight of 60 kDa, was detected in all HMMCL but became strongly labelled after neuraminidase treatment only in the HT cell lines. Thus, the expression of glycoproteins rich in terminal galactose residues may characterize human melanoma cells with different tumorigenic behavior.     

恶性淋巴瘤凝集素受体的研究

对32例恶性淋巴瘤(ML)的凝集素受体研究结果表明:ML组织中含有多种特异性糖蛋白,可被多种凝集素标记。刀豆凝集素(ConA),麦胚凝集素(WGA),豌豆凝集素(PSA),蓖麻凝集素(RCA)有较强的敏感性,可作为稳定可靠的标志。凝集素受体不仅有胞浆型、胞膜型定位,尚有核膜型定位。ConA受体在肿瘤组与非肿瘤对照组的核膜型定位上表现出很大的不同(P值<0.01),这具有实际应用意义。PSA的显色反应提示了该种凝集素受体的分布可能与淋巴瘤分化程度有关。RCA对R—S细胞的标记结果,不支持R—S细胞来源于组织细胞的学说。     

Lectin-binding sites in preneoplastic and neoplastic lesions of human and rodent respiratory tracts

The localization of fluorescein-labeled lectins, i.e., concanavalin A (Con A), Ricinus communis-120 (RCA), and wheat-germ agglutinin (WGA), were studied histologically in F344 rat epithelial lesions produced in the course of chemical carcinogenesis. WGA could not be demonstrated in these lesions. Although all lesions showed positive-binding sites when high concentrations of either Con A or RCA were used, a dilution study showed that the epithelial lesions had different affinities for lectins. With both Con A and RCA, dysplastic and neoplastic lesions showed the strongest intensity of fluorescence and squamous metaplasia showed the weakest. Normal and hyperplastic epithelia showed intermediate intensity. In the dilution study, RCA showed eight times more affinity and Con A showed two times more affinity for dysplastic and neoplastic epithelia than for normal or hyperplastic epithelium. Similar affinity patterns were observed in human lesions and tumors. With Con A, 58% of tumors showed much stronger fluorescence than did normal epithelium, and 44% of the tumors showed positive fluorescence with RCA. Although both lectins exhibited a stronger affinity for all the dysplastic-neoplastic lesions than for normal or hyperplastic epithelium, RCA proved to be the most adequate marker for preneoplastic lesions.     

Heterogeneity of human natural killer cells with respect to lectin-binding ability

The binding ability of peanut agglutinin (PNA), lentil agglutinin (LEN), soybean agglutinin (SBA), wheat germ agglutinin (WGA), and asparagus pea agglutinin (ASP) to human natural killer (NK) cells with the use of the double-marker immunofluorescence technique was studied. For identification of NK cells, VEP 13 (DC 16) monoclonal antibody was used. The receptor for PNA lectin was shown exclusively after neuraminidase treatment of cells, and VEP 13 antigen was neuraminidase resistant. The majority of VEP 13+ cells showed coexpression of lectin receptors for PNA, LEN, and WGA. Our results suggested that VEP 13 antigen and PNA receptor are two distinct membrane structures, whereas there is some competitive binding between LEN as well as WGA lectin and VEP 13 antibody. In double-marker experiments using PNA and LEN lectin, the small fraction of VEP 13+ cells lacking receptors for these lectins was found. In spite of neuraminidase treatment of the cells, no binding of SBA and ASP was shown. These results indicated apparent heterogeneity of NK cells with respect to lectin receptor expression. WGA lectin, which bound to all VEP 13+ cells, could probably be useful for isolation of NK cells.     

Binding of peanut agglutinin (PNA) and Dolichos biflorus agglutinin (DBA) to human colonic carcinomas, adenomas, and polyps: histochemical staining by the lectin-antilectin-PEP method

Binding of 2 lectins--peanut agglutinin (PNA) and Dolichos biflorus agglutinin (DBA)--with human colonic carcinomas, adenomas, juvenile and hyperplastic polyps, and polyps in Cronkhite-Canada syndrome was examined histochemically by the lectin-antilectin-PAP method. With PNA, brush borders of the carcinoma cell were stained(90.9%), while the cytoplasm of the adenomas and polyps were stained granularly (88.0%). With DBA, the cytoplasm of the carcinomas was stained diffusely (72.7%), though 25% of the cases showed the fringed type cytoplasmic staining similar to adenomas, polyps and normal crypts. It appears important to recognize the differences in the staining patterns, especially in PNA, between carcinomas and other benign lesions, which apparently reflect changes of sugar residues during cancerization.     

Lectin binding patterns in diffuse large cell lymphoma

The staining reaction of a panel of lectins in paraffin embedded lymph node specimens of diffuse large cell lymphoma was studied in relation to survival. In 47 of 49 patients, varying degrees of lectin binding were observed with Ricinus communis agglutinin (RCA), crude extract of Arachis hypogaea (c-PNA), Concanavalin ensiformis A (Con A), Triticum vulgaris A (WGA) and Phaseolus vulgaris A (PHA). Binding was either absent or only minimal with Pisum sativum A (PSA) and Lens culinaris A (LCA). Two categories of binding were observed: cell surface and cytoplasmic. Cell surface binding was seen in tumor cells, while cytoplasmic binding was observed in macrophage-histiocytes. Varying numbers of tumor cells were stained with RCA, WGA, c-PNA or PHA; but with Con A virtually no tumor cells were stained. Stromal macrophage-histiocytes were stained with RCA, WGA, or Con A in all but one case, frequently with all three lectins; c-PNA binding macrophage-histiocytes were absent in one third of the cases. With PHA the staining of stromal macrophage-histiocytes was extremely rare. Tumor cells that stained with RCA but not with c-PNA were observed in 9 of 15 patients who survived more than 2 years after diagnosis. In all 15 long-term survivors, stromal macrophage-histiocytes were positive for c-PNA. Tumor cells that reacted with c-PNA but not with RCA were seen in five patients who survived less than two years. All 16 patients whose tumors lacked c-PNA binding stromal macrophage-histiocytes in the presence of RCA binding macrophage-histiocytes were short-term survivors. These observations suggest the heterogeneity of stromal macrophage-histiocytes as well as that of tumor cells. Furthermore, the variation of lectin binding might be useful in assessing prognosis.     

Dual-parameter flow cytometry of transitional cell carcinomas. Quantitation of DNA content and binding of carbohydrate ligands in cellular subpopulations

Quantitative DNA measurements and estimates of blood group-related carbohydrate antigen expression have been used as predictive parameters in transitional cell carcinomas (Ca). To obtain an accurate quantitative characterization of cellular subpopulations on the basis of these parameters, the authors developed a dual-parameter flow cytometric method using a fluorescence-activated cell sorter. With this method single-cell suspensions from 26 transitional cell carcinomas were analyzed by means of propidium iodide (red fluorescence) as DNA ligand, and peanut agglutinin (PNA), wheat germ agglutinin (WGA), and anti-blood group A antibody (aBGA) as carbohydrate ligands. The latter ligands were visualized directly or indirectly by FITC (green fluorescence). The carbohydrate ligand binding was correlated to the DNA content of cell populations in the way that aneuploid populations showed a higher PNA binding (P less than 0.0002) and a lower WGA (P less than 0.01) and aBGA (P less than 0.04) binding than did diploid cell populations. The binding of PNA to aneuploid populations could be further increased (P less than 0.004) by neuraminidase treatments. Thus, aneuploid cells express both neuraminic acid substituted and unsubstituted PNA receptors. The carbohydrate ligand binding was cell cycle-dependent, as it was reduced (less than 0.008) in the G2-M phase. A low WGA (P less than 0.004) or aBGA (P less than 0.02) binding was correlated to tissue invasion. Immunohistochemistry with the carbohydrate ligands showed a good correlation between aBGA (P less than 0.0005) and PNA (P less than 0.004) binding to tumor cells and flow cytometric assay of these, as well as a correlation (P less than 0.003) between cellular location of WGA receptors and flow cytometric assay of these. It seems that dual-parameter flow cytometry represents an important tool in the characterization of bladder tumors.     

Lectin histochemistry of the thyroid gland

The authors carried out a histochemical study with lectins (Ulex europaeus agglutinin-I [UEA-I], Triticum vulgaris [WGA], Glycine max [SBA], Dolichos biflorus [DBA], and Arachis hypogaea [PNA]) in different thyroid gland conditions (17 benign nodular goiters, three diffuse hyperplasias, five Hashimoto's thyroiditis, 20 follicular adenomas, 14 well-differentiated papillary carcinomas, five well-differentiated follicular carcinomas, and 30 normal thyroids) in order to determine if specific lectin patterns are developed during neoplastic transformation. The results showed that (1) in normal thyroid glands, the lectin, UEA-I, is able to discriminate between follicular cells and C-cells; (2) pathologic follicular epithelium had an increased expression of UEA-I, SBA, and WGA receptors; (3) no lectin or group of lectins allow a distinction between follicular carcinoma and papillary carcinoma; (4) when benign and malignant tumors are compared for UEA-I affinity there is a significantly greater frequency of malignant tumour with UEA-I receptor; and (5) although all investigated lectins have shown receptors in endothelial cells at least in one case, the most constant findings have been obtained with UEA-I and WGA. These findings suggest that lectins are not useful in routine diagnostic pathologic examination; however, in particular cases of follicular carcinoma, UEA-I may be a useful tool for the recognition of small vessels invaded by tumoral cells and the demonstration of fucose residues in malignant tumor cells.     

Peanut lectin binding in breast carcinoma. Lack of correlation with estrogen receptor content

Previous reports have shown binding of peanut agglutinin (PNA) by immunoperoxidase techniques in normal, benign proliferative, and malignant breast epithelia. Correlation of binding with maturity, secretory activity, hormonal milieu, and tissue hormone receptor content has been described. To investigate the relationship between PNA staining by the avidin-biotin complex technique and estrogen receptor (ER) content, 79 breast tumors of different types and known tissue ER content were studied. Thirty-eight of 50 ER-positive cases were PNA-positive. Twenty of 29 ER-negative cases were PNA-positive. Statistical analysis shows the two factors to be independent (0.5 less than P less than 0.9). The literature on blood group antigens in breast carcinoma and histochemical applications of PNA is reviewed.     

Analyses of protein extracts of human breast cancers: changes in glycoprotein content linked to the malignant phenotype

The protein and glycoprotein composition of Triton X-100 extracts of breast biopsies from 17 women with benign breast disease and from 11 women with invasive breast carcinoma were investigated using electrophoresis in SDS-containing gradient polyacrylamide gels, followed by Coomassie Blue (CB) staining and the binding of radio-iodinated wheat germ agglutinin (WGA). Patterns were analysed after the CB-step for differences in protein composition, and after the WGA-step for differences in glycoprotein composition. Tissue extracts from patients with benign breast disease have less CB stained bands than similar extracts from the cancer patients. A particular consistent change was the appearance of an extra band at 58 Kdaltons in the cancer extracts. In contrast to the CB results, WGA detected less major bands, in the 40-60 Kd region, in the cancer extracts than at similar locations in benign extracts. Analysis of blood sera using the above techniques suggested that certain serum proteins could account for some of the WGA changes, but not the changes after CB staining. However, residual contamination of the specimens by blood proteins seemed unlikely because of the washing procedure used, unless these components were very strongly associated with the tissue. Differential synthesis of serum proteins by benign and malignant breast tissue may also explain some of our findings. Examination of the histopathology adjacent to the extracted tissue suggested that the degree of reduction in WGA-binding may be related to the extent of local invasiveness. Other animal and human studies suggest that reduced glycosylation of tumour-associated proteins may be linked to increased malignancy. The current findings may reflect a general pattern of change in tumour glycoprotein composition linked to malignant expression.