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Distinctive Features of Advancing Breast Cancer Cells and Interactions with Surrounding Stroma Observed Under the Scanning Electron Microscope

  • Jaafar, Hasnan (Department of Pathology, School of Medical Sciences, Health Campus, University Science Malaysia) ;
  • Sharif, Sharifah Emilia Tuan (Department of Pathology, School of Medical Sciences, Health Campus, University Science Malaysia) ;
  • Murtey, Mogana Das (Department of Pathology, School of Medical Sciences, Health Campus, University Science Malaysia)
  • Published : 2012.04.30

Abstract

Breast cancer cells undergo transformation when they spread into surrounding tissues. Studies have shown that cancer cells undergo surface alterations and interact with the surrounding microenvironment during the invasion process. The aim of the present study was to analyse these cancer cell surface alterations and interactions of cancer cells and stroma. Twenty 1-methyl-1-nitrosourea-induced breast cancer samples taken from five rats were fixed in McDowell-Trump fixative and then washed in 0.1 M phosphate buffer. The samples were then treated with osmium tetroxide before being washed in distilled water and subsequently dehydrated through graded ethanols. The dehydrated samples were immersed in hexamethyldisilazane (HMDS), then following removal of excess HMDS, the samples were air dried at room temperature in a dessicator. The dried samples were mounted onto specimen stubs and coated with gold coater before being viewed under a scanning electron microscope. We detected the presence of membrane ruffles on the surface of cancer cells and the formation of unique surface membrane protrusions to enhance movement and adhesion to the surrounding stroma during the process of invasion. Advancing cancer cells demonstrated formation of lamellipodia and invadopodia. The stroma at the advancing edge was desmoplastic with many collagen fibres laid down near the cancer cells. Our data suggest that all of these abnormalities could act as hallmarks of invasiveness for breast cancer.

Keywords

References

  1. Abbineni G, Modali S, Safiejko-Mroczka B, Petrenko V A, Mao C (2010). Evolutionary selection of new breast cancer celltargeting peptides and phages with the cell-targeting peptides fully displayed on the major coat and their effects on actin dynamics during cell internalization. Mol Pharm, 7, 1629-42. https://doi.org/10.1021/mp100052y
  2. Ada-Nguema A S, Xenias H, Hofman J M, et al (2006). The small GTPase R-Ras regulates organization of actin and drives membrane protrusions through the activity of PLCepsilon. J Cell Sci, 119, 1307-19. https://doi.org/10.1242/jcs.02835
  3. Araujo J C, Teran FC, Oliveira R A et al (2003). Comparison of hexamethyldisilazane and critical point drying treatments for SEM analysis of anaerobic biofilms and granular sludge. J Electron Microsc (Tokyo), 52, 429-33. https://doi.org/10.1093/jmicro/52.4.429
  4. Bailly M, Condeelis JS, Segall JE (1998). Chemoattractantinduced lamellipod extension. Microsc Res Tech, 43, 433-43. https://doi.org/10.1002/(SICI)1097-0029(19981201)43:5<433::AID-JEMT9>3.0.CO;2-2
  5. Benbow U, Schoenermark MP, Orndorff KA, Givan AL, Brinckerhoff CE (1999). Human breast cancer cells activate procollagenase-1 and invade type I collagen: invasion is inhibited by all-trans retinoic acid. Clin Exp Metastasis, 17, 231-8. https://doi.org/10.1023/A:1006639214618
  6. Brinkley B R, Beall P T, Wible LJ, et al (1980) Variations in cell form and cytoskeleton in human breast carcinoma cells in vitro. Cancer Res, 40, 3118-29.
  7. Dysktra MJ (1992). Biological Electron Microscopy, Theory, Techniques and Troubleshooting. Plenum Press NY, ?, 16-49.
  8. Huang J, Hardy JD, Sun Y, Shively JE (1999). Essential role of biliary glycoprotein (CD66a) in morphogenesis of the human mammary epithelial cell line MCF10F. J Cell Sci, 112, 4193-205.
  9. Jaafar H F, Idris M, Mohd Nafi S N, (2009). The association between phenotype and size of breast tumors induced by 1-methyl-1-nitrosourea (MNU) injection in rats. Med Sci Monit, 15, 129-34.
  10. Jiang P, Enomoto A, Takahashi M (2009). Cell biology of the movement of breast cancer cells: intracellular signalling and the actin cytoskeleton. Cancer Lett, 284, 122-30. https://doi.org/10.1016/j.canlet.2009.02.034
  11. Jing J, Tarbutton E, Wilson G, Prekeris R (2009). Rab11-FIP3 is a Rab11-binding protein that regulates breast cancer cell motility by modulating the actin cytoskeleton. Eur J Cell Biol, 88, 325-41. https://doi.org/10.1016/j.ejcb.2009.02.186
  12. Johnston C L, Cox H C, Gomm J J, Coombes R C (1995). bFGF and aFGF induce membrane ruffling in breast cancer cells but not in normal breast epithelial cells: FGFR-4 involvement. Biochem J, 306, 609-16.
  13. Kataoka S, Tsuruo T (1996). Physician Education: Apoptosis. Oncologist, 1, 399-401.
  14. Kaul-Ghanekar R, Singh S, Mamgain H, et al (2009). Tumor suppressor protein SMAR1 modulates the roughness of cell surface: combined AFM and SEM study. BMC Cancer, 9, 350. https://doi.org/10.1186/1471-2407-9-350
  15. Kelly T, Yan Y, Osborne RL, et al (1998). Proteolysis of extracellular matrix by invadopodia facilitates human breast cancer cell invasion and is mediated by matrix metalloproteinases. Clin Exp Metastasis, 16, 501-12. https://doi.org/10.1023/A:1006538200886
  16. Kusafuka K, Muramatsu K, Kasami M, et al (2008). Cartilaginous features in matrix-producing carcinoma of the breast: four cases report with histochemical and immunohistochemical analysis of matrix molecules. Mod Pathol, 21, 1282-92. https://doi.org/10.1038/modpathol.2008.120
  17. Kramer R H, Bensch KG, Wong J (1986). Invasion of reconstituted basement membrane matrix by metastatic human tumor cells. Cancer Res, 46, 1980-9.
  18. Luparello C, Sheterline P, Pucci-Minafra I, Minafra S (1991). A comparison of spreading and motility behaviour of 8701-BC breast carcinoma cells on type I, I-trimer and type V collagen substrata. Evidence for a permissive effect of type I-trimer collagen on cell locomotion. J Cell Sci, 100, 179-85.
  19. Martin S L, McDowell A, Lynas JF, Nelson J, Walker B (2001). A study of the anti-invasive properties of N-alphaphthalimidomethyl- ketomethylene tripeptide-based metalloprotease inhibitors. J Pharm Pharmacol, 53, 333-43. https://doi.org/10.1211/0022357011775569
  20. McHardy L M, Warabi K, Andersen R J, Roskelley C D, Roberge M (2005). Strongylophorine-26, a Rho-dependent inhibitor of tumor cell invasion that reduces actin stress fibers and induces nonpolarized lamellipodial extensions. Mol Cancer Ther, 4, 772-8. https://doi.org/10.1158/1535-7163.MCT-04-0310
  21. Meira DD, Marinho-Carvalho MM, Teixeira CA, et al (2004). Clotrimazole decreases human breast cancer cells viability through alterations in cytoskeleton-associated glycolytic enzymes. Molecular Genetics and Metabolism, 84, 354-62.
  22. Nikkhah M, Strobl JS, Peddi B (2009). Cytoskeletal role in differential adhesion patterns of normal fibroblasts and breast cancer cells inside silicon microenvironments. Biomed Microdevices, 11, 585-95. https://doi.org/10.1007/s10544-008-9268-2
  23. Provenzano P P, Eliceiri KW, Campbell JM, et al (2006). Collagen reorganization at the tumor-stromal interface facilitates local invasion. BMC Med, 4, 38. https://doi.org/10.1186/1741-7015-4-38
  24. Provenzano PP, Inman DR, Eliceiri K W, et al (2008) Collagen density promotes mammary tumor initiation and progression. BMC Med, 6, 11. https://doi.org/10.1186/1741-7015-6-11
  25. Rajah T T, Rambo DJ, Dmytryk JJ (2001). Influence of antiestrogens on NIH-3T3-fibroblast-induced motility of breast cancer cells. Chemotherapy, 47, 56-69. https://doi.org/10.1159/000048502
  26. Rizk N N, El-Rakhawy MT (1981). Tissue culture and scanning electron microscopy of breast carcinoma 'cell line MCF-7' from pleural effusion. Acta Anat (Basel), 109, 70-4. https://doi.org/10.1159/000145367
  27. Russo J, Tait L, Russo IH (1991). Morphological expression of cell transformation induced by c-Ha-ras oncogene in human breast epithelial cells. J Cell Sci, 99, 453-63.
  28. Sapino A, Pietribiasi F, Bussolati G, Marchisio P C (1986). Estrogen- and tamoxifen-induced rearrangement of cytoskeletal and adhesion structures in breast cancer MCF-7 cells. Cancer Res, 46, 2526-31.
  29. Schindler M, Ahmed I, Kamal J, et al (2005). A synthetic nanofibrillar matrix promotes in vivo-like organization and morphogenesis for cells in culture. Biomaterials, 26, 5624-31. https://doi.org/10.1016/j.biomaterials.2005.02.014
  30. Stoica G E, Franke T F, Moroni M, et al (2003). Effect of estradiol on estrogen receptor-alpha gene expression and activity can be modulated by the ErbB2/PI 3-K/Akt pathway. Oncogene, 22, 7998-8011. https://doi.org/10.1038/sj.onc.1206769
  31. Sugiura T, Berditchevski F (1999). Function of alpha3beta1- tetraspanin protein complexes in tumor cell invasion. Evidence for the role of the complexes in production of matrix metalloproteinase 2 (MMP-2). J Cell Biol, 146, 1375-89. https://doi.org/10.1083/jcb.146.6.1375
  32. Wang P S, Chou F S, Porchia L, Saji M, Pinzone J J (2008). Troglitazone inhibits cell migration, adhesion, and spreading by modulating cytoskeletal rearrangement in human breast cancer cells. Mol Carcinog, 47, 905-15. https://doi.org/10.1002/mc.20429
  33. Wolf K, Friedl P (2009). Mapping proteolytic cancer cellextracellular matrix interfaces. Clin Exp Metastasis, 26, 289-98. https://doi.org/10.1007/s10585-008-9190-2
  34. Xanthopoulos J M, Romano AE, Majumdar SK (2005). Response of Mouse Breast Cancer Cells to Anastrozole, Tamoxifen, and the Combination. J Biomed Biotechnol, 1, 10-9.
  35. Yamada H, Abe T, Li S, et al (2009). Dynasore, a dynamin inhibitor, suppresses lamellipodia formation and cancer cell invasion by destabilizing actin filaments. Biochem Biophys Res Commun, 390, 1142-8. https://doi.org/10.1016/j.bbrc.2009.10.105
  36. Yan J, Vassil A, Hait WN (2001). Involvement of phosphotidylinositol-3-kinase in membrane ruffling induced by P-glycoprotein substrates in multidrug-resistant carcinoma cells. Biochem Pharmacol, 63, 959-66.

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