Flow cytometric analysis of cell surface carbohydrates in metastatic human breast cancer

Alam, Sadaf; Whitford, P.; Cushley, William; George, W.D.; Campbell, Ailsa M.

doi:10.1038/bjc.1990.267

Cited by 40 publications

(25 citation statements)

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“…Leathem et al (1984Leathem et al ( , 1985Leathem et al ( , 1987 found that lectin from the albumin gland of the Roman snail, Helix pomatia agglutinin (HPA), which recognises N-acetyl-galactosaminyl residues, binds to a population of breast-cancer cells associated with AX metastases. Other investigators (Fenlon et al, 1987;Alam et al, 1990;Brooks & Leathem, 1991) also reported that the presence of HPA binding correlated with AX metastases, whereas it has also been reported (Fukutomi et al, 1989;Galea et al, 1991) Brooks et al (1991) and our may have some bearing on the significance of the HPA staining, although the incidence of HPA staining (45%) was similar with the incidence of AX metastases (46%) in this study. Since it is troublesome for clinicians to calculate the discriminant function for each patient, and one of the principal factors included in the discriminant function was clinical axillary status, however, the usefulness of the discriminant function would be limited.…”

Section: Discussioncontrasting

confidence: 55%

“…Lectins are proteins from different origins which bind to carbohydrate groups. Among these proteins, Brooks et al (1991) suggested that Helix pomatia agglutinin (HPA) might provide valuable information in breast cancer patients in whom AX dissection has not been preformed, because there is a significant relationship between the HPA staining and AX metastases (Leathem et al, 1984;Fenlon et al, 1987;Alam et al, 1990 Postoperatively, lymph nodes were removed from the resected specimens. The average total number of dissected axillary lymph nodes per patients was 26 nodes, with a range of 12-105.…”

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confidence: 99%

“…The second major site of regional metastases is the internal mammary lymph node (IM) chain. It is well known that histologic presence or absence of AX and/or IM is a crucial diagnostic parameter for the prognosis of the breast cancer patients (Veronesi et al, 1985; Noguchi et al, 1991 (Leathem et al, 1984;Fenlon et al, 1987;Alam et al, 1990 Postoperatively, lymph nodes were removed from the resected specimens. The average total number of dissected axillary lymph nodes per patients was 26 nodes, with a range of 12-105.…”

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confidence: 99%

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Further analysis of predictive value of Helix pomatia lectin binding to primary breast cancer for axillary and internal mammary lymph node metastases

Noguchi

Thomas²,

Kitagawa³

et al. 1993

Br J Cancer

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The axillary lymph node (AX) region is the principal site for metastatic spread from carcinoma of the breast. The second major site of regional metastases is the internal mammary lymph node (IM) chain. It is well known that histologic presence or absence of AX and/or IM is a crucial diagnostic parameter for the prognosis of the breast cancer patients (Veronesi et al., 1985; Noguchi et al., 1991 (Leathem et al., 1984;Fenlon et al., 1987;Alam et al., 1990 Postoperatively, lymph nodes were removed from the resected specimens. The average total number of dissected axillary lymph nodes per patients was 26 nodes, with a range of 12-105. At least three sections were made of each lymph node for histological examination with Hematoxylin and Eosin staining. There were 53 patients with negative AX, 20 with one to three positive AX, and 25 with four or more positive AX. IM were positive in 17 patients and negative in 81 patients.The method for preparation of the paraffin sections for HPA staining has been described elsewhere (Fukutomi et al., 1989). Most cases were clearly either intensively positive or completely negative. In a few cases we used the scoring defined by Brooks et al. (1991). Finally cases were classified as positive or negative staining.Statistically, a comparison was made using the chi-square test. In the univariate study, overall or disease-free survival was studied by the Kaplan-Meier method (Kaplan & Meier, 1958), and a log-rank test was used to assess statistical significance. In the multivariate study, Cox's regression test was used to examine several parameters simultaneously, followed by multiple regression analysis to determine which variables are important for predicting AX or IM metastases. From the coefficients of variables selected by a stepwise forward selection method, we were able to construct the following discriminant function. Discriminant score (Z) = a. + a,X, +. ..... + apXp, (a.; constant; al, a2.ap: discriminant coefficient; XI, X2. Xp: explanatory variables). The probability (P) of positive AX or IM was calculated by the following logistic function: P = ez/(1 + ez) = 1/(1 + e-z), e = 2.718.-). For predicting the AX or IM metastases, Z>0 (P>0.5) was regarded as positive AX or IM, and Z.0 (P<0.5) as negative AX or IM.Results Correspondence: M. Noguchi, Operation Center, Kanazawa University Hospital, School of Medicine, Kanazawa University, Takaramachi, 13-1, Kanazawa, 920, Japan. * Recipient of a fellowship from the Japanese-Germany Center, Berlin, Germany. Received 30 June 1992; and in revised form 6 January 1993.Forty-four (45%) of the 98 breast cancers were HPA-positive, and the remaining 54 (55%), HPA-negative. The positive rates of HPA staining were significantly higher as the disease progressed, with 7% for Stage 1, 42% for Stage 2, and 69% for Stage 3 (P<0.01). Moreover, HPA staining was found to be significantly associated with tumour size Br.

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Section: Discussioncontrasting

confidence: 55%

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confidence: 99%

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confidence: 99%

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Further analysis of predictive value of Helix pomatia lectin binding to primary breast cancer for axillary and internal mammary lymph node metastases

Noguchi

Thomas²,

Kitagawa³

et al. 1993

Br J Cancer

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“…The lectin from Helix pomatia (HPA) has emerged as an interesting marker of altered glycosylation in cancer, where expression of glycoconjugates recognised by the lectin appear to be associated with poor patient prognosis (see Mitchell and Schumacher, 1999;Brooks, 2000 for review). This association has been reported in breast cancer (Fenlon et al, 1987;Leathem and Brooks, 1987;Fukutomi et al, 1989Fukutomi et al, , 1991Alam et al, 1990;Noguchi et al, 1993aNoguchi et al, , 1993bNoguchi et al, , 1994Thomas et al, 1993), and also in adenocarcinomas at other common sites, including gastric cancer (Macartney, 1986;Kakeji et al, 1991;Maehara et al, 1995;Okuyama et al, 1998), carcinoma of the oesophagus (Yoshida et al, 1993(Yoshida et al, , 1994a colorectal carcinoma (Ikeda et al, 1994;Schumacher MD et al, 1994a, carcinoma of the thyroid (Sasano et al, 1989) and prostate cancer (Shiraishi et al, 1992). Only one published study has failed to confirm this relationship (Gusterson et al, 1993), and it appears that this may have been the fault of the inappropriate methodology employed .…”

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confidence: 80%

GalNAc glycoprotein expression by breast cell lines, primary breast cancer and normal breast epithelial membrane

2001

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Summary Over-expression of N-acetylgalactosamine glycoproteins as detected by binding of the lectin from Helix pomatia (HPA), is associated with metastatic competence and poor patient prognosis in a range of human adenocarcinomas. These glycoproteins remain poorly characterised, and their functional role has yet to be elucidated. This study describes characterisation of a range of human breast/breast cancer cell lines for the expression of the N-acetylgalactosaminylated glycoproteins of interest, and their comparison with normal breast epithelium and a range of clinical breast carcinoma samples. Confocal and light microscopy studies revealed cytochemical HPA-binding patterns consistent with a fundamental disruption in normal glycobiosynthetic pathways attending increasing metastatic potential. We report the most complete comparative analysis of HPA-binding ligands from cultured breast cells, clinical breast carcinoma samples and normal breast epithelium to date. Lectin blotting identified 11 major HPA-binding glycoprotein bands common to both clinical tumour samples and breast cell lines and 6 of these bands were also expressed by samples of normal breast epithelium, albeit at much lower levels. Moreover, very marked quantitative but not qualitative differences in levels of expression consistent with metastatic capability were noted.

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“…In particular, fluorescentlylabeled lectins, in combination with micro arrays and flow cytometry, have been used to analyze cell surface carbohydrates in several tissue cell lines [17,18]. In this current study, plant derived lectins were used to clarify the mechanism by which prebiotic oligosaccharides compete with cognate ligand in the surface of HEp-2 cells.…”

Section: Discussionmentioning

confidence: 99%

Inhibition of lectin adherence to tissue culture cells by prebiotic carbohydrates

Maldonado¹,

Fangman²,

Pinto³

et al. 2013

ABB

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Prebiotic carbohydrates, in addition to their ability to influence the colonic microbiota, are also able to inhibit attachment of pathogenic bacteria to epithelial cells. This effect is mediated via their structural similarity to the carbohydrate ligands, located on the mucosal cell surface to which bacterial lectins attach. However, the mechanism for this inhibition is not well understood. The goal of this research was to measure the effect of two prebiotic carbohydrates, galactooligosaccharide and polydextrose, on the binding kinetics of plant lectins, having known ligand specificity, to tissue culture cells. To measure adherence, competetion experiments were conducted with HEp-2 cells exposed to nine fluorescent-labeled lectins and either the cognate ligand or a prebiotic. Fluorescence microscopy and image analysis were used to quantify adherence. Lectins that were able to bind to target cells were significantly inhibited in the presence of the cognate ligands. When prebiotics were added, inhibittion of lectin binding was observed, depending on the structural similarity between the prebiotic and the cognate ligands. In general, PDX did not inhibit lectin attachment, whereas GOS significantly inhibited most lectins. This research suggests that receptor sites located on the surface of epithelial HEp-2 cells are structurally similar to GOS.

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Flow cytometric analysis of cell surface carbohydrates in metastatic human breast cancer

Cited by 40 publications

References 10 publications

Further analysis of predictive value of Helix pomatia lectin binding to primary breast cancer for axillary and internal mammary lymph node metastases

Further analysis of predictive value of Helix pomatia lectin binding to primary breast cancer for axillary and internal mammary lymph node metastases

GalNAc glycoprotein expression by breast cell lines, primary breast cancer and normal breast epithelial membrane

Inhibition of lectin adherence to tissue culture cells by prebiotic carbohydrates

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