Skip to main content
British Journal of Cancer logoLink to British Journal of Cancer
. 2001 Sep;85(7):1014–1022. doi: 10.1054/bjoc.2001.2028

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

S A Brooks 1, D M S Hall 1, I Buley 2
PMCID: PMC2375105  PMID: 11592774

Abstract

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. © 2001 Cancer Research Campaignhttp://www.bjcancer.com

Keywords: Helix pomatia lectin, HPA, GalNAc glycoproteins, breast cancer, metastasis, cell lines

Full Text

The Full Text of this article is available as a PDF (143.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alam S. M., Whitford P., Cushley W., George W. D., Campbell A. M. Flow cytometric analysis of cell surface carbohydrates in metastatic human breast cancer. Br J Cancer. 1990 Aug;62(2):238–242. doi: 10.1038/bjc.1990.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Briand P., Petersen O. W., Van Deurs B. A new diploid nontumorigenic human breast epithelial cell line isolated and propagated in chemically defined medium. In Vitro Cell Dev Biol. 1987 Mar;23(3):181–188. doi: 10.1007/BF02623578. [DOI] [PubMed] [Google Scholar]
  3. Brooks S. A., Carter T. M. N-acetylgalactosamine, N-acetylglucosamine and sialic acid expression in primary breast cancers. Acta Histochem. 2001 Feb;103(1):37–51. doi: 10.1078/0065-1281-00576. [DOI] [PubMed] [Google Scholar]
  4. Brooks S. A., Leathem A. J., Camplejohn R. S., Gregory W. Markers of prognosis in breast cancer--the relationship between binding of the lectin HPA and histological grade, SPF, and ploidy. Breast Cancer Res Treat. 1993;25(3):247–256. doi: 10.1007/BF00689839. [DOI] [PubMed] [Google Scholar]
  5. Brooks S. A., Leathem A. J. Expression of alpha-GalNAc glycoproteins by breast cancers. Br J Cancer. 1995 May;71(5):1033–1038. doi: 10.1038/bjc.1995.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brooks S. A., Leathem A. J. Prediction of lymph node involvement in breast cancer by detection of altered glycosylation in the primary tumour. Lancet. 1991 Jul 13;338(8759):71–74. doi: 10.1016/0140-6736(91)90071-v. [DOI] [PubMed] [Google Scholar]
  7. Brooks S. A., Lymboura M., Schumacher U., Leathem A. J. Histochemistry to detect Helix pomatia lectin binding in breast cancer: methodology makes a difference. J Histochem Cytochem. 1996 May;44(5):519–524. doi: 10.1177/44.5.8627008. [DOI] [PubMed] [Google Scholar]
  8. Brooks S. A. The involvement of Helix pomatia lectin (HPA) binding N-acetylgalactosamine glycans in cancer progression. Histol Histopathol. 2000 Jan;15(1):143–158. doi: 10.14670/HH-15.143. [DOI] [PubMed] [Google Scholar]
  9. Cailleau R., Young R., Olivé M., Reeves W. J., Jr Breast tumor cell lines from pleural effusions. J Natl Cancer Inst. 1974 Sep;53(3):661–674. doi: 10.1093/jnci/53.3.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Calafat J., Janssen H. Binding of lectins to human mammary tumors: ultrastructural study. Breast Cancer Res Treat. 1984;4(3):169–179. doi: 10.1007/BF01806482. [DOI] [PubMed] [Google Scholar]
  11. Engel L. W., Young N. A., Tralka T. S., Lippman M. E., O'Brien S. J., Joyce M. J. Establishment and characterization of three new continuous cell lines derived from human breast carcinomas. Cancer Res. 1978 Oct;38(10):3352–3364. [PubMed] [Google Scholar]
  12. Fenlon S., Ellis I. O., Bell J., Todd J. H., Elston C. W., Blamey R. W. Helix pomatia and Ulex europeus lectin binding in human breast carcinoma. J Pathol. 1987 Jul;152(3):169–176. doi: 10.1002/path.1711520305. [DOI] [PubMed] [Google Scholar]
  13. Filmus J., Pollak M. N., Cailleau R., Buick R. N. MDA-468, a human breast cancer cell line with a high number of epidermal growth factor (EGF) receptors, has an amplified EGF receptor gene and is growth inhibited by EGF. Biochem Biophys Res Commun. 1985 Apr 30;128(2):898–905. doi: 10.1016/0006-291x(85)90131-7. [DOI] [PubMed] [Google Scholar]
  14. Fukutomi T., Hirohashi S., Tsuda H., Nanasawa T., Yamamoto H., Itabashi M., Shimosato Y. The prognostic value of tumor-associated carbohydrate structures correlated with gene amplifications in human breast carcinomas. Jpn J Surg. 1991 Sep;21(5):499–507. doi: 10.1007/BF02470985. [DOI] [PubMed] [Google Scholar]
  15. Fukutomi T., Itabashi M., Tsugane S., Yamamoto H., Nanasawa T., Hirota T. Prognostic contributions of Helix pomatia and carcinoembryonic antigen staining using histochemical techniques in breast carcinomas. Jpn J Clin Oncol. 1989 Jun;19(2):127–134. [PubMed] [Google Scholar]
  16. Hammarström S., Kabat E. A. Purification and characterization of a blood-group A reactive hemagglutinin from the snail Helix pomatia and a study of its combining site. Biochemistry. 1969 Jul;8(7):2696–2705. doi: 10.1021/bi00835a002. [DOI] [PubMed] [Google Scholar]
  17. Ikeda Y., Mori M., Adachi Y., Matsushima T., Sugimachi K. Prognostic value of the histochemical expression of helix pomatia agglutinin in advanced colorectal cancer. A univariate and multivariate analysis. Dis Colon Rectum. 1994 Feb;37(2):181–184. doi: 10.1007/BF02047545. [DOI] [PubMed] [Google Scholar]
  18. Kakeji Y., Tsujitani S., Mori M., Maehara Y., Sugimachi K. Helix pomatia agglutinin binding activity is a predictor of survival time for patients with gastric carcinoma. Cancer. 1991 Dec 1;68(11):2438–2442. doi: 10.1002/1097-0142(19911201)68:11<2438::aid-cncr2820681119>3.0.co;2-#. [DOI] [PubMed] [Google Scholar]
  19. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  20. Laidler P., Lityńska A. Tumor cell N-glycans in metastasis. Acta Biochim Pol. 1997;44(2):343–357. [PubMed] [Google Scholar]
  21. Laitinen L., Juusela H., Virtanen I. Binding of the blood group-reactive lectins to human adult kidney specimens. Anat Rec. 1990 Jan;226(1):10–17. doi: 10.1002/ar.1092260103. [DOI] [PubMed] [Google Scholar]
  22. Langlois A. J., Holder W. D., Jr, Iglehart J. D., Nelson-Rees W. A., Wells S. A., Jr, Bolognesi D. P. Morphological and biochemical properties of a new human breast cancer cell line. Cancer Res. 1979 Jul;39(7 Pt 1):2604–2613. [PubMed] [Google Scholar]
  23. Lasfargues E. Y., Coutinho W. G., Redfield E. S. Isolation of two human tumor epithelial cell lines from solid breast carcinomas. J Natl Cancer Inst. 1978 Oct;61(4):967–978. [PubMed] [Google Scholar]
  24. Leathem A. J., Brooks S. A. Predictive value of lectin binding on breast-cancer recurrence and survival. Lancet. 1987 May 9;1(8541):1054–1056. doi: 10.1016/s0140-6736(87)90482-x. [DOI] [PubMed] [Google Scholar]
  25. Macartney J. C. Lectin histochemistry of galactose and N-acetyl-galactosamine glycoconjugates in normal gastric mucosa and gastric cancer and the relationship with ABO and secretor status. J Pathol. 1986 Oct;150(2):135–144. doi: 10.1002/path.1711500208. [DOI] [PubMed] [Google Scholar]
  26. Maehara Y., Okuyama T., Kakeji Y., Endo K., Yamamoto M., Sugimachi K. A tumour-associated cell-surface glycoprotein accompanying p53 overexpression and higher growth potential for gastric cancer. Br J Cancer. 1995 May;71(5):999–1002. doi: 10.1038/bjc.1995.192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Mitchell B. S., Schumacher U. The use of the lectin Helix pomatia agglutinin (HPA) as a prognostic indicator and as a tool in cancer research. Histol Histopathol. 1999 Jan;14(1):217–226. doi: 10.14670/HH-14.217. [DOI] [PubMed] [Google Scholar]
  28. Mitchell B. S., Vernon K., Schumacher U., Habil M. Ultrastructural localization of Helix pomatia agglutinin (HPA)-binding sites in human breast cancer cell lines and characterization of HPA-binding glycoproteins by western blotting. Ultrastruct Pathol. 1995 Jan-Feb;19(1):51–59. doi: 10.3109/01913129509014603. [DOI] [PubMed] [Google Scholar]
  29. Noguchi M., Thomas M., Kitagawa H., Kinishita K., Kinami S., Takamura H., Miyazaki I., Mizukami Y. DNA ploidy and helix pomatia lectin binding as predictors of regional lymph node metastases and prognostic factors in breast cancer. Breast Cancer Res Treat. 1993;26(1):67–75. doi: 10.1007/BF00682701. [DOI] [PubMed] [Google Scholar]
  30. Noguchi M., Thomas M., Kitagawa H., Kinoshita K., Ohta N., Nagamori M., Miyazaki I. Further analysis of predictive value of Helix pomatia lectin binding to primary breast cancer for axillary and internal mammary lymph node metastases. Br J Cancer. 1993 Jun;67(6):1368–1371. doi: 10.1038/bjc.1993.253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Okuyama T., Maehara Y., Kakeji Y., Tsuijitani S., Korenaga D., Sugimachi K. Interrelation between tumor-associated cell surface glycoprotein and host immune response in gastric carcinoma patients. Cancer. 1998 Apr 15;82(8):1468–1475. doi: 10.1002/(sici)1097-0142(19980415)82:8<1468::aid-cncr6>3.0.co;2-5. [DOI] [PubMed] [Google Scholar]
  32. Roth J. Cytochemical localization of terminal N-acetyl-D-galactosamine residues in cellular compartments of intestinal goblet cells: implications for the topology of O-glycosylation. J Cell Biol. 1984 Feb;98(2):399–406. doi: 10.1083/jcb.98.2.399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sasano H., Rojas M., Silverberg S. G. Analysis of lectin binding in benign and malignant thyroid nodules. Arch Pathol Lab Med. 1989 Feb;113(2):186–189. [PubMed] [Google Scholar]
  34. Schumacher U., Adam E., Brooks S. A., Leathem A. J. Lectin-binding properties of human breast cancer cell lines and human milk with particular reference to Helix pomatia agglutinin. J Histochem Cytochem. 1995 Mar;43(3):275–281. doi: 10.1177/43.3.7868857. [DOI] [PubMed] [Google Scholar]
  35. Schumacher U., Adam E., Flavell D. J., Boehm D., Brooks S. A., Leathem A. J. Glycosylation patterns of the human colon cancer cell line HT-29 detected by Helix pomatia agglutinin and other lectins in culture, in primary tumours and in metastases in SCID mice. Clin Exp Metastasis. 1994 Nov;12(6):398–404. doi: 10.1007/BF01755883. [DOI] [PubMed] [Google Scholar]
  36. Schumacher U., Higgs D., Loizidou M., Pickering R., Leathem A., Taylor I. Helix pomatia agglutinin binding is a useful prognostic indicator in colorectal carcinoma. Cancer. 1994 Dec 15;74(12):3104–3107. doi: 10.1002/1097-0142(19941215)74:12<3104::aid-cncr2820741207>3.0.co;2-0. [DOI] [PubMed] [Google Scholar]
  37. Shiraishi T., Atsumi S., Yatani R. Comparative study of prostatic carcinoma bone metastasis among Japanese in Japan and Japanese Americans and whites in Hawaii. Adv Exp Med Biol. 1992;324:7–16. doi: 10.1007/978-1-4615-3398-6_2. [DOI] [PubMed] [Google Scholar]
  38. Soule H. D., Vazguez J., Long A., Albert S., Brennan M. A human cell line from a pleural effusion derived from a breast carcinoma. J Natl Cancer Inst. 1973 Nov;51(5):1409–1416. doi: 10.1093/jnci/51.5.1409. [DOI] [PubMed] [Google Scholar]
  39. Streets A. J., Brooks S. A., Dwek M. V., Leathem A. J. Identification, purification and analysis of a 55 kDa lectin binding glycoprotein present in breast cancer tissue. Clin Chim Acta. 1996 Oct 15;254(1):47–61. doi: 10.1016/0009-8981(96)06363-2. [DOI] [PubMed] [Google Scholar]
  40. Thomas M., Noguchi M., Fonseca L., Kitagawa H., Kinoshita K., Miyazaki I. Prognostic significance of Helix pomatia lectin and c-erbB-2 oncoprotein in human breast cancer. Br J Cancer. 1993 Sep;68(3):621–626. doi: 10.1038/bjc.1993.397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Virtanen I. Helix pomatia agglutinin binds specifically to the Golgi apparatus in cultured human fibroblasts and reveals two Golgi apparatus-specific glycoproteins. Histochemistry. 1990;94(4):397–401. doi: 10.1007/BF00266446. [DOI] [PubMed] [Google Scholar]
  43. Wessel D., Flügge U. I. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem. 1984 Apr;138(1):141–143. doi: 10.1016/0003-2697(84)90782-6. [DOI] [PubMed] [Google Scholar]
  44. Yoshida Y., Okamura T., Shirakusa T. An immunohistochemical study of helix pomatia agglutinin binding on carcinomas of the esophagus. Surg Gynecol Obstet. 1993 Sep;177(3):299–302. [PubMed] [Google Scholar]
  45. Yoshida Y., Okamura T., Yano K., Ezaki T. Silver stained nucleolar organizer region proteins and Helix pomatia agglutinin immunostaining in esophageal carcinoma: correlated prognostic factors. J Surg Oncol. 1994 Jun;56(2):116–121. doi: 10.1002/jso.2930560215. [DOI] [PubMed] [Google Scholar]
  46. Zak I., Lewandowska E., Gnyp W. Selectin glycoprotein ligands. Acta Biochim Pol. 2000;47(2):393–412. [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES