Abstract
The detection of occult metastatic breast cancer cells by RT-PCR is limited by the poor specificity of most tumour mRNA markers. MAGE-A3 is a highly specific tumour mRNA marker that is not expressed in non-cancer cells. This study assesses MAGE-A3 mRNA as a molecular marker for the detection of tumour cells in the sentinel lymph nodes (SLN) of breast cancer patients. Serial frozen sections of SLN (n= 121) were obtained from 77 AJCC (American Joint Committee on Cancer) Stage I–IIIA breast cancer patients. MAGE-A3 mRNA analysis of SLN was performed by RT-PCR and Southern blot analysis. Tumour cells were detected in 48 of 121 (40%) SLN from 77 patients by H&E or IHC staining, and 35 of 77 (45%) patients, overall, had histopathologically (H&E and/or IHC) positive SLN. Among histopathologically negative SLN, 28 of 73 (38%) SLN were MAGE-A3 mRNA positive by RT-PCR. Overall, 41 of 77 (53%) patients and 50 of 121 (41%) SLN were positive for MAGE-A3. MAGE-A3 mRNA expression in the SLN occurred more frequently with infiltrating lobular carcinoma (P< 0.001) than with infiltrating ductal carcinoma, adding further evidence of possible phenotypic differences between these 2 subtypes of breast cancer. Due to its high specificity, MAGE-A3 mRNA is a potentially useful marker for detecting breast cancer cells in the SLN. One half of breast tumours expressed MAGE-A3 mRNA, which has important potential implications for antigen-specific targeted immunotherapy. © 2001 Cancer Research Campaign http://www.bjcancer.com
Similar content being viewed by others
Article PDF
Change history
16 November 2011
This paper was modified 12 months after initial publication to switch to Creative Commons licence terms, as noted at publication
References
Barth A, Craig PH and Silverstein MJ (1997) Predictors of axillary lymph node metastases in patients with T1 breast carcinoma. Cancer 79: 1918–1922
Bentz JS, Yassa N and Clayton F (1998) Pleomorphic lobular carcinoma of the breast: clinicopathologic features of 12 cases. Mod Pathol 11: 814–822
Bostick PJ, Chatterjee S, Chi DD, Huynh KT, Giuliano AE, Cote R and Hoon DS (1998a) Limitations of specific reverse-transcriptase polymerase chain reaction markers in the detection of metastases in the lymph nodes and blood of breast cancer patients. J Clin Oncol 16: 2632–2640
Bostick PJ, Hoon DS and Cote RJ (1998b) Detection of carcinoembryonic antigen messenger RNA in lymph nodes from patients with colorectal cancer [letter; comment]. N Engl J Med 339: 1643–1644
Bostick PJ, Huynh KT, Sarantou T, Turner RR, Qi K, Giuliano AE and Hoon DS (1998c) Detection of metastases in sentinel lymph nodes of breast cancer patients by multiple-marker RT-PCR. Int J Cancer 79: 645–651
Bostick PJ, Morton DL, Turner RR, Huynh KT, Wang HJ, Elashoff R, Essner R and Hoon DS (1999) Prognostic Significance of Occult Metastases Detected by Sentinel Lymphadenectomy and Reverse Transcriptase-Polymerase Chain Reaction in Early-Stage Melanoma Patients. J Clin Oncol 17: 3238–3244
Brugger W, Buhring HJ, Grunebach F, Vogel W, Kaul S, Muller R, Brummendorf TH, Ziegler BL, Rappold I, Brossart P, Scheding S and Kanz L (1999) Expression of MUC-1 epitopes on normal bone marrow: implications for the detection of micrometastatic tumor cells. J Clin Oncol 17: 1535–1544
Burchill SA, Bradbury MF, Pittman K, Southgate J, Smith B and Selby P (1995) Detection of epithelial cancer cells in peripheral blood by reverse transcriptase-polymerase chain reaction. Br J Cancer 71: 278–281
Carter BA, Jensen RA, Simpson JF and Page DL (2000) Benign transport of breast epithelium into axillary lymph nodes after biopsy. Am J Clin Pathol 113: 259–265
Chi DD, Merchant RE, Rand R, Conrad AJ, Garrison D, Turner R, Morton DL and Hoon DS (1997) Molecular detection of tumor-associated antigens shared by human cutaneous melanomas and gliomas. Am J Pathol 150: 2143–2152
Datta YH, Adams PT, Drobyski WR, Ethier SP, Terry VH and Roth MS (1994) Sensitive detection of occult breast cancer by the reverse-transcriptase polymerase chain reaction. J Clin Oncol 12: 475–482
Finke J, Fritzen R, Ternes P, Lange W and Dolken G (1993) An improved strategy and a useful housekeeping gene for RNA analysis from formalin-fixed, paraffin-embedded tissues by PCR. Biotechniques 14: 448–453
Fitzgibbons PL, Page DL, Weaver D, Thor AD, Allred DC, Clark GM, Ruby SG, O’Malley F, Simpson JF, Connolly JL, Hayes DF, Edge SB, Lichter A and Schnitt SJ (2000) Prognostic factors in breast cancer. College of American Pathologists Consensus Statement 1999. Arch Pathol Lab Med 124: 966–978
Fujie T, Mori M, Ueo H, Sugimachi K and Akiyoshi T (1997) Expression of MAGE and BAGE genes in Japanese breast cancers. Ann Oncol 8: 369–372
Gaugler B, Van den Eynde B, van der Bruggen P, Romero P, Gaforio JJ, De Plaen E, Lethe B, Brasseur F and Boon T (1994) Human gene MAGE-3 codes for an antigen recognized on a melanoma by autologous cytolytic T lymphocytes. J Exp Med 179: 921–930
Giuliano AE, Kirgan DM, Guenther JM and Morton DL (1994) Lymphatic mapping and sentinel lymphadenectomy for breast cancer. Ann Surg 220: 391–398
Giuliano AE, Dale PS, Turner RR, Morton DL, Evans SW and Krasne DL (1995) Improved axillary staging of breast cancer with sentinel lymphadenectomy. Ann Surg 222: 394–399
Giuliano AE, Jones RC, Brennan M and Statman R (1997) Sentinel lymphadenectomy in breast cancer. J Clin Oncol 15: 2345–2350
Hammond ME, Fitzgibbons PL, Compton CC, Grignon DJ, Page DL, Fielding LP, Bostwick D and Pajak TF (2000) College of American Pathologists Conference XXXV: solid tumor prognostic factors-which, how and so what? Summary document and recommendations for implementation. Cancer Committee and Conference Participants. Arch Pathol Lab Med 124: 958–965
Hellman S (1994) Karnofsky Memorial Lecture: Natural history of small breast cancers. J Clin Oncol 12: 2229–2234
Hoon DS, Doi F, Giuliano AE, Schmid P and Conrad AJ (1995a) The detection of breast carcinoma micrometastases in axillary lymph nodes by means of reverse transcriptase-polymerase chain reaction [letter]. Cancer 76: 533–535
Hoon DS, Wang Y, Dale PS, Conrad AJ, Schmid P, Garrison D, Kuo C, Foshag LJ, Nizze AJ and Morton DL (1995b) Detection of occult melanoma cells in blood with a multiple-marker polymerase chain reaction assay. J Clin Oncol 13: 2109–2116
Hoon DS, Yuzuki D, Hayashida M and Morton DL (1995c) Melanoma patients immunized with melanoma cell vaccine induce antibody responses to recombinant MAGE-1 antigen. J Immunol 154: 730–737
Hoon DS, Sarantou T, Doi F, Chi DD, Kuo C, Conrad AJ, Schmid P, Turner R and Guiliano A (1996) Detection of metastatic breast cancer by beta-hCG polymerase chain reaction. Int J Cancer 69: 369–374
International (Ludwig) Breast Cancer Study Group (1990) Prognostic importance of occult axillary lymph node micrometastases from breast cancers. Lancet 335: 1565–1568
Lambrechts AC, Bosma AJ, Klaver SG, Top B, Perebolte L, van’t Veer LJ and Rodenhuis S (1999) Comparison of immunocytochemistry, reverse transcriptase polymerase chain reaction, and nucleic acid sequence-based amplification for the detection of circulating breast cancer cells. Breast Cancer Res Treat 56: 219–231
Lee AH, Dublin EA, Bobrow LG and Poulsom R (1998) Invasive lobular and invasive ductal carcinoma of the breast show distinct patterns of vascular endothelial growth factor expression and angiogenesis. J Pathol 185: 394–401
Moinfar F, Man YG, Lininger RA, Bodian C and Tavassoli FA (1999) Use of keratin 35betaE12 as an adjunct in the diagnosis of mammary intraepithelial neoplasia-ductal type—benign and malignant intraductal proliferations. Am J Surg Pathol 23: 1048–1058
Mori M, Mimori K, Inoue H, Barnard GF, Tsuji K, Nanbara S, Ueo H and Akiyoshi T (1995) Detection of cancer micrometastases in lymph nodes by reverse transcriptase-polymerase chain reaction. Cancer Res 55: 3417–3420
Mori M, Inoue H, Mimori K, Shibuta K, Baba K, Nakashima H, Haraguchi M, Tsuji K, Ueo H, Barnard GF and Akiyoshi T (1996) Expression of MAGE genes in human colorectal carcinoma. Ann Surg 224: 183–188
Neville AM, Bettelheim R, Gelber RD, Save-Soderbergh J, Davis BW, Reed R, Torhorst J, Golouh R, Peterson HF and Price KN et al (1992) Factors predicting treatment responsiveness and prognosis in node-negative breast cancer. The International (Ludwig) Breast Cancer Study Group. J Clin Oncol 10: 696–705
Noguchi S, Aihara T, Motomura K, Inaji H, Imaoka S and Koyama H (1996) Detection of breast cancer micrometastases in axillary lymph nodes by means of reverse transcriptase-polymerase chain reaction. Comparison between MUC1 mRNA and keratin 19 mRNA amplification. Am J Pathol 148: 649–656
Novaes M, Bendit I, Garicochea B and del Giglio A (1997) Reverse transcriptase-polymerase chain reaction analysis of cytokeratin 19 expression in the peripheral blood mononuclear cells of normal female blood donors. Mol Pathol 50: 209–211
Oaks MK, Hanson JP Jr. and O’Malley DP (1994) Molecular cytogenetic mapping of the human melanoma antigen (MAGE) gene family to chromosome region Xq27-qter: implications for MAGE immunotherapy. Cancer Res 54: 1627–1629
Okamoto T, Kaneda Y, Yuzuki D, Huang SK, Chi DD and Hoon DS (1997) Induction of antibody response to human tumor antigens by gene therapy using a fusigenic viral liposome vaccine. Gene Ther 4: 969–976
Pantel K, Cote RJ and Fodstad O (1999) Detection and clinical importance of micrometastatic disease. J Natl Cancer Inst 91: 1113–1124
Prasad ML, Osborne MP and Hoda SA (1998) Observations on the histopathologic diagnosis of microinvasive carcinoma of the breast. Anat Pathol 3: 209–232
Reyes CV, Thompson KS, Jensen JD and Choudhury AM (1998) Metastasis of unknown origin: the role of fine-needle aspiration cytology. Diagn Cytopathol 18: 319–322
Richard F, Pacyna-Gengelbach M, Schlüns K, Fleige B, Winzer KJ, Szymas J, Dietel M, Petersen I and Schwendel A (2000) Patterns of chromosomal imbalances in invasive breast cancer. Int J Cancer 89: 305–310
Russo V, Traversari C, Verrecchia A, Mottolese M, Natali PG and Bordignon C (1995) Expression of the MAGE gene family in primary and metastatic human breast cancer: implications for tumor antigen-specific immunotherapy. Int J Cancer 64: 216–221
Schoenfeld A, Luqmani Y, Smith D, O’Reilly S, Shousha S, Sinnett HD and Coombes RC (1994) Detection of breast cancer micrometastases in axillary lymph nodes by using polymerase chain reaction. Cancer Res 54: 2986–2990
Sinha PS, Bendall S and Bates T (2000) Does routine grading of invasive lobular cancer of the breast have the same prognostic significance as for ductal cancers?. Eur J Surg Oncol 26: 733–737
Thurner B, Haendle I, Roder C, Dieckmann D, Keikavoussi P, Jonuleit H, Bender A, Maczek C, Schreiner D, von den Driesch P, Brocker EB, Steinman RM, Enk A, Kampgen E and Schuler G (1999) Vaccination with mage-3A1 peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma. J Exp Med 190: 1669–1678
Toikkanen S, Pylkkanen L and Joensuu H (1997) Invasive lobular carcinoma of the breast has better short-and long-term survival than invasive ductal carcinoma. Br J Cancer 76: 1234–1240
Turner RR, Ollila DW, Krasne DL and Giuliano AE (1997) Histopathologic validation of the sentinel lymph node hypothesis for breast carcinoma. Ann Surg 226: 271–276
Turner RR, Ollila DW, Stern S and Giuliano AE (1999) Optimal histopathologic examination of the sentinel lymph node for breast carcinoma staging [see comments]. Am J Surg Pathol 23: 263–267
Van der Bruggen P, Traversari C, Chomez P, Lurquin C, De Plaen E, Van den Eynde B, Knuth A and Boon T (1991) A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma. Science 254: 1643–1647
Weber JS, Hua FL, Spears L, Marty V, Kuniyoshi C and Celis E (1999) A phase I trial of an HLA-A1 restricted MAGE-3 epitope peptide with incomplete Freund’s adjuvant in patients with resected high-risk melanoma. J Immunother 22: 431–440
Xu X, Roberts SA, Pasha TL and Zhang PJ (2000) Undesirable cytokeratin immunoreactivity of native nonepithelial cells in sentinel lymph nodes from patients with breast carcinoma. Arch Pathol Lab Med 124: 1310–1313
Yun K, Gunn J, Merrie AE, Phillips LV and McCall JL (1997) Keratin 19 mRNA is detectable by RT-PCR in lymph nodes of patients without breast cancer [letter; comment]. Br J Cancer 76: 1112–1113
Zeng Z, Melamed J, Symmans PJ, Cangiarella JF, Shapiro RL, Peralta H and Symmans WF (1999) Benign proliferative nipple duct lesions frequently contain CAM 5.2 and anti-cytokeratin 7 immunoreactive cells in the overlying epidermis. Am J Surg Pathol 23: 1349–1355
Zippelius A, Kufer P, Honold G, Kollermann MW, Oberneder R, Schlimok G, Riethmuller G and Pantel K (1997) Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow. J Clin Oncol 15: 2701–2708
Author information
Authors and Affiliations
Rights and permissions
From twelve months after its original publication, this work is licensed under the Creative Commons Attribution-NonCommercial-Share Alike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
About this article
Cite this article
Wascher, R., Bostick, P., Huynh, K. et al. Detection of MAGE-A3 in breast cancer patients’ sentinel lymph nodes. Br J Cancer 85, 1340–1346 (2001). https://doi.org/10.1054/bjoc.2001.2079
Received:
Revised:
Accepted:
Published:
Issue date:
DOI: https://doi.org/10.1054/bjoc.2001.2079
Keywords
This article is cited by
-
Melanoma associated antigen (MAGE)-A3 promotes cell proliferation and chemotherapeutic drug resistance in gastric cancer
Cellular Oncology (2016)
-
ABCG2 is a potential marker of tumor-initiating cells in breast cancer
Tumor Biology (2015)
-
The melanoma-associated antigen-A3, -A4 genes: relation to the risk and clinicopathological parameters in breast cancer patients
Molecular and Cellular Biochemistry (2011)
-
Sentinel lymph node mapping for primary breast cancer
Current Oncology Reports (2005)
-
Detection of lymph node metastases of papillary thyroid cancer—comparison of the results of histopathology, immunohistochemistry and reverse transcription-polymerase chain reaction—a preliminary report
Langenbeck's Archives of Surgery (2005)


