Abstract
Purpose
Brain metastases are an increasingly common complication in breast cancer patients. The Metastasis Suppressor Genes (MSG) Nm23, KISS1, KAI1, BRMS1, and Mkk4 have been associated with the metastatic potential of breast cancer in vitro and in vivo.
Methods
The mRNA expression of Nm23, KISS1, KAI1, BRMS1, and Mkk4 in fresh frozen tissue samples of brain metastases from ductal invasive breast cancer specimens was examined in relation to primary tumors. In a first step, mRNA expression screening was carried out using a semi-quantitative RT-PCR approach, in a second step quantitative real-time RT-PCR was performed on selected specimens. By immunohistochemical staining, gene products were visualized on the protein level.
Results
Semi-quantitative RT-PCR revealed reduced mRNA expression of Nm23, KISS1, KAI1, BRMS, and Mkk4 in brain metastases. Results for KISS1, KAI1, BRMS, and Mkk4 were confirmed by real-time RT-PCR. In detail, mRNA expression reduction in breast cancer brain metastases was tenfold. Expression of MSG could be confirmed by immunohistochemical staining on protein level.
Conclusions
Our investigations revealed significantly reduced mRNA expression of metastases suppressor genes KISS1, KAI1, BRMS1, and Mkk4 in breast cancer brain metastasis. Particularly, in the case of KISS1 and Mkk4, an important role for future treatment of patients with breast cancer brain metastatic lesions can be assumed.
Similar content being viewed by others
References
Adachi M, Taki T, Leki Y, Huang CL, Higashiyama M, Miyake M (1996) Correlation of Kai1/CD82 gene expression with good prognosis in patients with non-small cell lung cancer. Cancer Res 56:1751–1755
Ahmad A, Hart IR (1997) Mechanism of metastasis. Crit Rev Oncol Hematol 26:163–173
Arbit E, Wronski M (1997) Clinical decision making in brain metastases. Neurosurg Clin N Am 7:447–457
Arnaud-Dabernat S, Bourbon PM, Dierich A, Le Meur M, Daniel JY (2003) Knockout mice as model system for studying Nm23/NDP kinase gene functions. Application to the Nm23-M1 gene. J Bioenerg Biomembr 35:19–30
Cavallaro U, Christofori G (2001) Cell adhesion in tumor invasion and metastasis: loss of the glue is not enough. BBA Rev Cancer 1552:39–45
Chekmareva MA, Kadkhodaian MM, Hollowell CM, Kim H, Yosdida BA, Luu HB, Stadler WM, Rinker-Schaeffer CW (1998) Chromosome 12-mediated dormancy of AT6.1 prostate cancer micrometastases. Cancer Res 58:4963–4969
Clayton AJ, Danson S, Jolly S, Ryder WD, Burt PA, Stewart AL, Wilkinson PM, Welch RS, Magee B, Wilson G, Howell A, Wardley AM (2004) Incidence of cerebral metastases in patients treated with trastuzumab for metastatic breast cancer. Br J Cancer [Epub ahead of print]
Cuenda A (2000) Mitogen-activated protein kinase kinase 4 (MKK4) Int J Biochem Cell Biol 32:581–587
Debies MT, Welch DR (2002) Genetic basis of human breast cancer metastasis. J Mamm Gliand Biol Neopl 6:441–451
Dong JT, Isaacs WB, Barrett JC, Isaacs JT (1997) Genomic organization of the human KAI1 metastasis-suppressor gene. Genomics 41:25–32
Dursun A, Akyurek N, Gunel N, Yamac D (2002) Prognostic implication of Nm23-H1 expression in colorectal carcinoma. Pathology 34:427–432
Dusonchet L, Corsale S, Migliavacca M, Calo V, Bazan V, Amato A, Cammareri P, Totaro MS, Agnese V, Cascio S, La Rocca G, Sisto PS, Dardanoni G, Valerio MR, Grassi N, Latteri S, Cajozzo M, Buscemi M, Castorina S, Morello V, Tomasino RM, Gebbia N, Russo A (2003) Nm23-H1 expression does not predict clinical survival in colorectal cancer patients. Oncol Rep 10:1257–1263
Freije JM, MacDonald NJ, Steeg PS (1998) Nm23 and tumour metastasis: basic and translational advances. Biochem Soc Symp 63:261–271
Harms JF, Welch DR, Miele ME (2003) KISS1 metastasis suppression and emergent pathways. Clin Exp Metastasis 20:11–18
Heide I, Thiede C, Poppe K, de Kant E, Huhn D, Rochlitz C (1994) Expression and mutational analysis of Nm23-H1 in liver metastases of colorectal cancer. Br J Cancer 70:1267–1271
Heimann R, Ferguson D, Hellmann S (1998) The relationship between Nm23, angiogenesis, and the metastatic proclivity of node-negative breast cancer. Cancer Res 58:2766–2771
Howlett AR, Petersen OW, Steeg PS, Bissell MJ (1994) A novel function for Nm23: overexpression in human brast carcinoma cells leads to the formation of basement membrane and growth arrest. J Natl Cancer Inst 86:1838–2844
Jackson P, Puisieux A (2000) Is the KAI1 metastasis suppressor gene a cellular target of p53? A review of current evidence. Biochem Biophys Res Commun 278:499–502
Kim HL, Vander Griend DJ, Yang X, Benson DA, Dubauskas Z, Yoshida BA, Chekmareva MA, Ichikawa Y, Sokoloff MH, Zhan P, Karrison T, Lin A, Stadler WM, Ichikawa T, Rubin MA, Rinker-Schaeffer CW (2001) Mitogen-activated protein kinase kinase 4 metastasis suppressor gene expression is inversely related to histological pattern in advancing human prostatic cancers. Cancer Res 61:2833–1837
Kleihues P, Cavenee WK (2000) Pathology and genetics of tumours of the nervous system. IARC, Lyon
Kotani M, Detheux M, Vandenbogaerde A, Communi D, Vanderwinden JM, Le Poul E, Brezillon S, Tyldesley R, Suarez-Huerta N, Vandeput F, Blanpain C, Schiffmann SN, Vassart G, Parmentier M (2001) The metastasis suppressor gene KiSS-1 encodes kisspeptin, the natural ligand of the orphan G protein-coupled receptor GPR54. J Biol Chem 276:34631–34636
Lee JH, Welch DR (1997) Suppression of metastasis in human breast carcinoma MDA-MB-435 cells after transfection with the metastasis suppressor gene, KiSS1. Cancer Res 57:2384–2387
Leone A, Flatow U, van Houtte K, Steeg PS (1993) Transfection of human Nm23-H1 into the human MDA-MB-435 breast carcinoma cell line: effects on tumor metatstic potential, colonization, and enzymatic activity. Oncogene 8:2325–2333
Liu H, Mao H, Fu X (2001) Expression of Nm23 in breast cancer—correlation with distant metastasis and prognosis. Zhonghua Zhong Liu Za Zhi 23:224–227
Liu L, Wu DH, Li ZG, Yang GZ, Ding YQ (2003) Effects of Kai1/CD82 on biological behaviour of human colorectal carcinoma cell line. World J Gastroenterol 9:231–1236
Lombardi DP, Geradts J, Foley JF, Chiao C, Lamb PW, Barrett JC (1999) Loss of KAI1 expression in pregression of colorectal cancer. Cancer Res 59:5724–5731
MacDonald NJ, de la Rosa A, Steeg PS (1995) The potential roles of Nm23 in cancer metastasis and cellular differentiation. Eur J Cancer 31A:1096–1100
Mao H, Liu H, Fu X, Fang Z, Abrams J, Worsham MJ (2001) Loss of Nm23 expression predicts distal metastases and poorer survival for breast cancer. Int J Oncol 18:587–591
Mashimoto T, Wanabe M, Hirota S, Hosobe S, Miura K, Tegtmeyer PJ, Rinker-Shaeffer CW, Watabe K (1998) The expression of the Kai1 gene, a tumor metastasis suppressor, is directly activated by P53. Proc Natl Acad Sci USA 95:11307–11311
Maurer CA, Graber HU, Firess H, Beyermann B, Willi D, Netzer P, Zimmermann A, Buchler MW (1999) Reduced expression of the metastasis suppressor gene Kai1 in advanced colon cancer and its metastases. Surgery 126:869–880
Meehan WJ, Welch DR (2003) Breast cancer metastasis suppressor gene 1: update. Clin Exp Metastasis 20:45–50
Meyer T, Hart IR (1998) Mechanisms of tumour metastasis. Eur J Cancer 34:214–221
Paget S (1889) The distribution of secondary growths in cancer of the breast. Lancet 1:571–573
Philips KK, White AE, Hicks DJ, Welch DR, Barrett JC, Wei LL, Weissman BE (1998) Correlation between reduction of metastasis in the MDA-MB-435 model system and increased expression of the KAI1 protein. Mol Carcinog 21:111–120
Puduvalli VK (2001) Brain metastases: biology and the role of the brain microenvironment. Curr Oncol Rep 3:467–475
Robinson VL, Hickson JA, Vander Griend DJ, Dubauskas Z, Rinker-Schaeffer CW (2003) MKK4 and metastasis suppression: a marriage of signal transduction and metastasis research. Clin Exp Metastasis 20:25–30
Saunders MM, Seraj MJ, Zhongyong L, Zhou Z, Winter CR, Welch DR, Donahue HJ (2001) Breast cancer metastatic potential correlates with a breakdown in homospecific and heterospecific gap junctional intercellular communication. Cancer Res 61:1765–1767
Seraj MJ, Samant RS, Verderame MF, Welch DR (2000) Functional evidence for a novel human breast carcinoma metastasis suppressor, BRMS1, encoded at chromosome 11q13. Cancer Res 60:2764–2769
Shevde LA, Samant RS, Goldberg SF, Sikaneta T, Alessandrini A, Donahue HJ, Mauger DT, Welch DR (2002) Suppression of human melanoma metastasis by the metastasis suppressor gene, BRMS1. Exp Cell Res 273:229–239
Shinohara T, Nishimura N, Hanibuchi M, Nokihara H, Miki T, Hamada H, Sone S (2001) Transduction of KAI1/CD82 cDNA promotes hematogenous spread of human lung-cancer cells in natural killer cell-depleted SCID mice. Int J Cancer 94:16–23
Shirasaki F, Takata M, Hatta N, Takehara K (2001) Loss of expression of the metastasis suppressor gene KiSS1 during melanoma progression and its association with LOH of chromosome 6q16.3-q23. Cancer Res 61:7422–7425
Steeg PS, Bevilacqua G, Kopper L, Thorgeirsson UP, Talmadge JE, Liotta LA,Sobel ME (1998) Evidence for a novel gene associated with low tumor metastatic potential. J Natl Cancer Inst 80:200–204
Wang L, Pan Y, Dai JL (2004) Evidence of Mkk4 pro-oncogenic activity in breast and pancreatic tumors. Oncogene [Epub ahead of print]
Welch DR, Steeg PS, Rinker-Schaeffer CW (2000) Molecular biology of breast cancer metastasis—Genetic regulation of human breast carcinoma metastasis. Breast Cancer Res 2:408–416
Wronski M, Arbit E, McCormick B (1997) Surgical treatment of 70 patients with brain metastases from breast carcinoma. Cancer 80:1746–54
Yamada SD, Hickson JA, Hrobowski Y, Vander Griend DJ, Benson D, Montag A, Karrison T, Huo D, Rutgers J, Adams S, Rinker-Schaeffer CW (2002) Mitigen-activated protein kinase kinase 4 (MKK4) acts as a metastasis suppressor gene in human ovarian carcinoma. Cancer Res 62:6717–6723
Yang X, Welch DR, Phillips KK, Weissman BE, Wei LL (1997) KAI1, a putative marker for metastatic potential in human breast cancer. Cancer Lett 119:149–155
Yang X, Wei L, Tang C, Slack R, Montgomery E, Lippman M (2000) Kai1 protein is down-regulated during the progression of human breast cancer. Clin Cancer Res 6:3424–3429
Yang X, Wei L, Tang C, Slack R, Mueller S, Lippman ME (2001) Overexpression of KAI1 suppresses in vitro invasiveness and in vivo metastasis in breast cancer cells. Cancer Res 61:284–5288
Yoshida BA, Dubauskas Z, Chekmareva MA, Christiano TR, Stadler WM, Rinker-Schaeffer CW (1999) Mitogen-activated protein kinase kinase 4/stress-activated protein/Erk kinase 1 (MKK4/SEK1), a prostate cancer metastasis suppressor gene encoded by human chromosome 17. Cancer Res 59:5483–5487
Acknowledgement
We are indebted to Frank Rösel, Department of Obstetrics and Gynecology University of Schleswig-Holstein Medical Center, Campus Kiel for the handling of tumor specimen. We also thank Brigitte Rehmke, Neurosurgical Research Laboratories, Dept. of Neurosurgery, University of Schleswig-Holstein Medical Center, Campus Kiel, for her technical assistance
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Stark, A.M., Tongers, K., Maass, N. et al. Reduced metastasis-suppressor gene mRNA-expression in breast cancer brain metastases. J Cancer Res Clin Oncol 131, 191–198 (2005). https://doi.org/10.1007/s00432-004-0629-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-004-0629-9