Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1998 Sep;78(6):710–717. doi: 10.1038/bjc.1998.566

Relationship of nm23 to proteolytic factors, proliferation and motility in breast cancer tissues and cell lines.

R L Russell 1, A N Pedersen 1, J Kantor 1, K Geisinger 1, R Long 1, N Zbieranski 1, A Townsend 1, B Shelton 1, N Brünner 1, T E Kute 1
PMCID: PMC2062960  PMID: 9743288

Abstract

Low expression of the antimetastatic gene nm23 has been associated with shorter overall survival in breast cancer. To better understand the mechanism(s) of action of this protein, we compared the levels of the nm23 protein in 152 breast cancer samples with other factors known to be involved in metastasis or related to prognosis. There was no significant relationship between either of the nm23 isoforms and cathepsin D (Cat-D), urokinase plasminogen activator (uPA), its inhibitor (PAI-1), steroid hormone receptors or ploidy status. A marginal inverse correlation was observed between per cent S-phase and nm23-H1 expression (r = -0.193, P = 0.047) and a positive correlation was observed between uPA receptor (uPAR) and both nm23-H1 (r = 0.263, P = 0.0018) and nm23-H2 (r = 0.230, P = 0.0064). The nm23-H1 gene was transfected into MDA-MB-231 human breast cancer cells and 12 clones were selected, of which two were characterized extensively. We found no significant differences in Cat-D, uPA, PAI-1 or uPAR, as a function of nm23 expression in either the MDA-MB-231 cells or the transfected clones. Compared with the parent cell line, we did observe a dose-dependent decrease in growth factor-stimulated motility and a decrease in metastatic potential in two clones with four- and eightfold elevated nm23-H1 expression, whereas the proliferative activities were similar. We conclude that the decreased metastatic potential might be related to down-regulation of growth factor-stimulated motility.

Full text

PDF
710

Selected References

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

  1. Barnes D. M., Harris W. H., Smith P., Millis R. R., Rubens R. D. Immunohistochemical determination of oestrogen receptor: comparison of different methods of assessment of staining and correlation with clinical outcome of breast cancer patients. Br J Cancer. 1996 Nov;74(9):1445–1451. doi: 10.1038/bjc.1996.563. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barnes R., Masood S., Barker E., Rosengard A. M., Coggin D. L., Crowell T., King C. R., Porter-Jordan K., Wargotz E. S., Liotta L. A. Low nm23 protein expression in infiltrating ductal breast carcinomas correlates with reduced patient survival. Am J Pathol. 1991 Aug;139(2):245–250. [PMC free article] [PubMed] [Google Scholar]
  3. Brünner N., Thompson E. W., Spang-Thomsen M., Rygaard J., Danø K., Zwiebel J. A. lacZ transduced human breast cancer xenografts as an in vivo model for the study of invasion and metastasis. Eur J Cancer. 1992;28A(12):1989–1995. doi: 10.1016/0959-8049(92)90245-w. [DOI] [PubMed] [Google Scholar]
  4. Bunting K. D., Townsend A. J. De novo expression of transfected human class 1 aldehyde dehydrogenase (ALDH) causes resistance to oxazaphosphorine anti-cancer alkylating agents in hamster V79 cell lines. Elevated class 1 ALDH activity is closely correlated with reduction in DNA interstrand cross-linking and lethality. J Biol Chem. 1996 May 17;271(20):11884–11890. doi: 10.1074/jbc.271.20.11884. [DOI] [PubMed] [Google Scholar]
  5. Caligo M. A., Cipollini G., Fiore L., Calvo S., Basolo F., Collecchi P., Ciardiello F., Pepe S., Petrini M., Bevilacqua G. NM23 gene expression correlates with cell growth rate and S-phase. Int J Cancer. 1995 Mar 16;60(6):837–842. doi: 10.1002/ijc.2910600619. [DOI] [PubMed] [Google Scholar]
  6. Campbell P. G., Novak J. F., Yanosick T. B., McMaster J. H. Involvement of the plasmin system in dissociation of the insulin-like growth factor-binding protein complex. Endocrinology. 1992 Mar;130(3):1401–1412. doi: 10.1210/endo.130.3.1371448. [DOI] [PubMed] [Google Scholar]
  7. Christensen L., Wiborg Simonsen A. C., Heegaard C. W., Moestrup S. K., Andersen J. A., Andreasen P. A. Immunohistochemical localization of urokinase-type plasminogen activator, type-1 plasminogen-activator inhibitor, urokinase receptor and alpha(2)-macroglobulin receptor in human breast carcinomas. Int J Cancer. 1996 May 16;66(4):441–452. doi: 10.1002/(SICI)1097-0215(19960516)66:4<441::AID-IJC6>3.0.CO;2-W. [DOI] [PubMed] [Google Scholar]
  8. Duffy M. J., Reilly D., O'Sullivan C., O'Higgins N., Fennelly J. J., Andreasen P. Urokinase-plasminogen activator, a new and independent prognostic marker in breast cancer. Cancer Res. 1990 Nov 1;50(21):6827–6829. [PubMed] [Google Scholar]
  9. Freije J. M., Blay P., MacDonald N. J., Manrow R. E., Steeg P. S. Site-directed mutation of Nm23-H1. Mutations lacking motility suppressive capacity upon transfection are deficient in histidine-dependent protein phosphotransferase pathways in vitro. J Biol Chem. 1997 Feb 28;272(9):5525–5532. doi: 10.1074/jbc.272.9.5525. [DOI] [PubMed] [Google Scholar]
  10. Gilles A. M., Presecan E., Vonica A., Lascu I. Nucleoside diphosphate kinase from human erythrocytes. Structural characterization of the two polypeptide chains responsible for heterogeneity of the hexameric enzyme. J Biol Chem. 1991 May 15;266(14):8784–8789. [PubMed] [Google Scholar]
  11. Goodall R. J., Dawkins H. J., Robbins P. D., Hähnel E., Sarna M., Hähnel R., Papadimitriou J. M., Harvey J. M., Sterrett G. F. Evaluation of the expression levels of nm23-H1 mRNA in primary breast cancer, benign breast disease, axillary lymph nodes and normal breast tissue. Pathology. 1994 Oct;26(4):423–428. doi: 10.1080/00313029400169122. [DOI] [PubMed] [Google Scholar]
  12. Grøndahl-Hansen J., Christensen I. J., Rosenquist C., Brünner N., Mouridsen H. T., Danø K., Blichert-Toft M. High levels of urokinase-type plasminogen activator and its inhibitor PAI-1 in cytosolic extracts of breast carcinomas are associated with poor prognosis. Cancer Res. 1993 Jun 1;53(11):2513–2521. [PubMed] [Google Scholar]
  13. Grøndahl-Hansen J., Peters H. A., van Putten W. L., Look M. P., Pappot H., Rønne E., Dano K., Klijn J. G., Brünner N., Foekens J. A. Prognostic significance of the receptor for urokinase plasminogen activator in breast cancer. Clin Cancer Res. 1995 Oct;1(10):1079–1087. [PubMed] [Google Scholar]
  14. Han S., Yun I. J., Noh D. Y., Choe K. J., Song S. Y., Chi J. G. Abnormal expression of four novel molecular markers represents a highly aggressive phenotype in breast cancer. Immunohistochemical assay of p53, nm23, erbB-2, and cathepsin D protein. J Surg Oncol. 1997 May;65(1):22–27. doi: 10.1002/(sici)1096-9098(199705)65:1<22::aid-jso5>3.0.co;2-q. [DOI] [PubMed] [Google Scholar]
  15. Hennessy C., Henry J. A., May F. E., Westley B. R., Angus B., Lennard T. W. Expression of the antimetastatic gene nm23 in human breast cancer: an association with good prognosis. J Natl Cancer Inst. 1991 Feb 20;83(4):281–285. doi: 10.1093/jnci/83.4.281. [DOI] [PubMed] [Google Scholar]
  16. Howlett A. R., Petersen O. W., Steeg P. S., Bissell M. J. A novel function for the nm23-H1 gene: overexpression in human breast carcinoma cells leads to the formation of basement membrane and growth arrest. J Natl Cancer Inst. 1994 Dec 21;86(24):1838–1844. doi: 10.1093/jnci/86.24.1838. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ji L., Arcinas M., Boxer L. M. The transcription factor, Nm23H2, binds to and activates the translocated c-myc allele in Burkitt's lymphoma. J Biol Chem. 1995 Jun 2;270(22):13392–13398. doi: 10.1074/jbc.270.22.13392. [DOI] [PubMed] [Google Scholar]
  18. Jänicke F., Schmitt M., Pache L., Ulm K., Harbeck N., Höfler H., Graeff H. Urokinase (uPA) and its inhibitor PAI-1 are strong and independent prognostic factors in node-negative breast cancer. Breast Cancer Res Treat. 1993;24(3):195–208. doi: 10.1007/BF01833260. [DOI] [PubMed] [Google Scholar]
  19. Kantor J. D., McCormick B., Steeg P. S., Zetter B. R. Inhibition of cell motility after nm23 transfection of human and murine tumor cells. Cancer Res. 1993 May 1;53(9):1971–1973. [PubMed] [Google Scholar]
  20. Kute T. E., Grøndahl-Hansen J., Shao S. M., Long R., Russell G., Brünner N. Low cathepsin D and low plasminogen activator type 1 inhibitor in tumor cytosols defines a group of node negative breast cancer patients with low risk of recurrence. Breast Cancer Res Treat. 1998 Jan;47(1):9–16. doi: 10.1023/a:1005882520982. [DOI] [PubMed] [Google Scholar]
  21. Kute T. E., Shao Z. M., Sugg N. K., Long R. T., Russell G. B., Case L. D. Cathepsin D as a prognostic indicator for node-negative breast cancer patients using both immunoassays and enzymatic assays. Cancer Res. 1992 Oct 1;52(19):5198–5203. [PubMed] [Google Scholar]
  22. Leone A., Flatow U., King C. R., Sandeen M. A., Margulies I. M., Liotta L. A., Steeg P. S. Reduced tumor incidence, metastatic potential, and cytokine responsiveness of nm23-transfected melanoma cells. Cell. 1991 Apr 5;65(1):25–35. doi: 10.1016/0092-8674(91)90404-m. [DOI] [PubMed] [Google Scholar]
  23. Leone A., Flatow U., VanHoutte K., Steeg P. S. Transfection of human nm23-H1 into the human MDA-MB-435 breast carcinoma cell line: effects on tumor metastatic potential, colonization and enzymatic activity. Oncogene. 1993 Sep;8(9):2325–2333. [PubMed] [Google Scholar]
  24. Liotta L. A., Steeg P. S., Stetler-Stevenson W. G. Cancer metastasis and angiogenesis: an imbalance of positive and negative regulation. Cell. 1991 Jan 25;64(2):327–336. doi: 10.1016/0092-8674(91)90642-c. [DOI] [PubMed] [Google Scholar]
  25. Lyons R. M., Gentry L. E., Purchio A. F., Moses H. L. Mechanism of activation of latent recombinant transforming growth factor beta 1 by plasmin. J Cell Biol. 1990 Apr;110(4):1361–1367. doi: 10.1083/jcb.110.4.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. MacDonald N. J., Freije J. M., Stracke M. L., Manrow R. E., Steeg P. S. Site-directed mutagenesis of nm23-H1. Mutation of proline 96 or serine 120 abrogates its motility inhibitory activity upon transfection into human breast carcinoma cells. J Biol Chem. 1996 Oct 11;271(41):25107–25116. doi: 10.1074/jbc.271.41.25107. [DOI] [PubMed] [Google Scholar]
  27. Ossowski L. Invasion of connective tissue by human carcinoma cell lines: requirement for urokinase, urokinase receptor, and interstitial collagenase. Cancer Res. 1992 Dec 15;52(24):6754–6760. [PubMed] [Google Scholar]
  28. Postel E. H. NM23/Nucleoside diphosphate kinase as a transcriptional activator of c-myc. Curr Top Microbiol Immunol. 1996;213(Pt 2):233–252. doi: 10.1007/978-3-642-61109-4_11. [DOI] [PubMed] [Google Scholar]
  29. Rochefort H. Cathepsin D in breast cancer. Breast Cancer Res Treat. 1990 Jul;16(1):3–13. doi: 10.1007/BF01806570. [DOI] [PubMed] [Google Scholar]
  30. Rosenquist C., Thorpe S. M., Danø K., Grøndahl-Hansen J. Enzyme-linked immunosorbent assay of urokinase-type plasminogen activator (uPA) in cytosolic extracts of human breast cancer tissue. Breast Cancer Res Treat. 1993 Dec;28(3):223–229. doi: 10.1007/BF00666583. [DOI] [PubMed] [Google Scholar]
  31. Royds J. A., Stephenson T. J., Rees R. C., Shorthouse A. J., Silcocks P. B. Nm23 protein expression in ductal in situ and invasive human breast carcinoma. J Natl Cancer Inst. 1993 May 5;85(9):727–731. doi: 10.1093/jnci/85.9.727. [DOI] [PubMed] [Google Scholar]
  32. Russell R. L., Geisinger K. R., Mehta R. R., White W. L., Shelton B., Kute T. E. nm23--relationship to the metastatic potential of breast carcinoma cell lines, primary human xenografts, and lymph node negative breast carcinoma patients. Cancer. 1997 Mar 15;79(6):1158–1165. doi: 10.1002/(sici)1097-0142(19970315)79:6<1158::aid-cncr14>3.0.co;2-z. [DOI] [PubMed] [Google Scholar]
  33. Rønne E., Høyer-Hansen G., Brünner N., Pedersen H., Rank F., Osborne C. K., Clark G. M., Danø K., Grøndahl-Hansen J. Urokinase receptor in breast cancer tissue extracts. Enzyme-linked immunosorbent assay with a combination of mono- and polyclonal antibodies. Breast Cancer Res Treat. 1995 Mar;33(3):199–207. doi: 10.1007/BF00665944. [DOI] [PubMed] [Google Scholar]
  34. Sastre-Garau X., Lacombe M. L., Jouve M., Véron M., Magdelénat H. Nucleoside diphosphate kinase/NM23 expression in breast cancer: lack of correlation with lymph-node metastasis. Int J Cancer. 1992 Feb 20;50(4):533–538. doi: 10.1002/ijc.2910500406. [DOI] [PubMed] [Google Scholar]
  35. Sato Y., Rifkin D. B. Inhibition of endothelial cell movement by pericytes and smooth muscle cells: activation of a latent transforming growth factor-beta 1-like molecule by plasmin during co-culture. J Cell Biol. 1989 Jul;109(1):309–315. doi: 10.1083/jcb.109.1.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sawan A., Lascu I., Veron M., Anderson J. J., Wright C., Horne C. H., Angus B. NDP-K/nm23 expression in human breast cancer in relation to relapse, survival, and other prognostic factors: an immunohistochemical study. J Pathol. 1994 Jan;172(1):27–34. doi: 10.1002/path.1711720107. [DOI] [PubMed] [Google Scholar]
  37. Stahl J. A., Leone A., Rosengard A. M., Porter L., King C. R., Steeg P. S. Identification of a second human nm23 gene, nm23-H2. Cancer Res. 1991 Jan 1;51(1):445–449. [PubMed] [Google Scholar]
  38. Steeg P. S., Bevilacqua G., Kopper L., Thorgeirsson U. P., Talmadge J. E., Liotta L. A., Sobel M. E. Evidence for a novel gene associated with low tumor metastatic potential. J Natl Cancer Inst. 1988 Apr 6;80(3):200–204. doi: 10.1093/jnci/80.3.200. [DOI] [PubMed] [Google Scholar]
  39. Stracke M. L., Liotta L. A. Multi-step cascade of tumor cell metastasis. In Vivo. 1992 Jul-Aug;6(4):309–316. [PubMed] [Google Scholar]
  40. Venturelli D., Martinez R., Melotti P., Casella I., Peschle C., Cucco C., Spampinato G., Darzynkiewicz Z., Calabretta B. Overexpression of DR-nm23, a protein encoded by a member of the nm23 gene family, inhibits granulocyte differentiation and induces apoptosis in 32Dc13 myeloid cells. Proc Natl Acad Sci U S A. 1995 Aug 1;92(16):7435–7439. doi: 10.1073/pnas.92.16.7435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. de la Rosa A., Williams R. L., Steeg P. S. Nm23/nucleoside diphosphate kinase: toward a structural and biochemical understanding of its biological functions. Bioessays. 1995 Jan;17(1):53–62. doi: 10.1002/bies.950170111. [DOI] [PubMed] [Google Scholar]

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

RESOURCES