Skip to main content

Advertisement

SpringerLink
Log in

Non-small-cell lung carcinoma cells invade human bronchial mucosa In vitro

  • Growth, Differentiation And Senescence
  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Summary

To study invasion of lung cancer in vitro a novel three-dimensional coculture assay consisting of living human tissues has been developed. Multicellular spheroids initiated from a new large-cell lung carcinoma cell line (GaL23), found to be invasive in immunodeficient mice, were confronted with precultured bronchial fragments derived from mucosal biopsies obtained during routine fiberoptic bronchoscopy. The bronchial fragments consist of a stromal core with scattered fibroblasts covered by a continuous surface epithelium resting on a basal lamina. During the first 2 wk of confrontation, a gradual retraction of the bronchial epithelium with subsequent adhesion of the tumor cells to the underlying basal lamina occurred. The following week, a limited invasion of tumor cells into the bronchial stroma was seen. To facilitate the entrance of tumor cells through the mucosal surface, the surface epithelium was removed prior to coculture by ethylenediaminetetraacetic acid (EDTA) buffer treatment. Upon confrontation, GaL23 cells then rapidly attached to and migrated on the exposed basal lamina and an increasing number of tumor cells was seen in the stroma during the first week of culture. This model offers opportunities for studying mechanisms of lung cancer adhesion, migration, and invasion using human bronchial mucosa as the natural target tissue.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. The world health organization histological typing of lung tumors. Am. J. Clin. Pathol. 77:123–136; 1982.

    Google Scholar 

  2. Alberts, B.; Bray, D.; Lewis, J., et al. Molecular biology of the cell. 3rd ed. New York: Garland; 1994.

    Google Scholar 

  3. Baba, M.; Klein-Szanto, A. J. P.; Trono, D., et al. Preneoplastic and neoplastic growth of xenotransplanted lung-derived human cell lines using deepithelialized rat tracheas. Cancer Res. 47:573–578; 1987.

    PubMed  CAS  Google Scholar 

  4. Bjerkvig, R., ed. Spheroid culture in cancer research. Boca Raton: CRC Press; 1992.

    Google Scholar 

  5. Booth, C.; Schriven, S. J.; Harnden, P., et al. An in vitro experimental model for the study of tumor invasion in transitional cell carcinoma of the bladder. In: Proceedings of the 88th Annual Meeting of the American Association for Cancer Research. San Diego, CA; 1997.

  6. de Ridder, L.; Mareel, M.; Vakaet, L. Adhesion of malignant and nonmalignant cells to cultured embryonic substrates. Cancer Res. 35:3164–3171; 1975.

    PubMed  Google Scholar 

  7. de Ridder, L.; Mareel, M.; Vakaet, L. Invasion of malignant cells into cultured embryonic substrates. Arch. Geschwulstforsch. 47:7–27; 1977.

    PubMed  Google Scholar 

  8. de Vries, J. E.; Dinjens, W. N.; De Bruyne, G. K., et al. In vivo and in vitro invasion in relation to phenotypic characteristics of human colorectal carcinoma cells. Br. J. Cancer 71:271–277; 1995.

    PubMed  Google Scholar 

  9. Dingemans, K. P.; Mooi, W. J. Invasion of lung tissue by bronchogenic squamous-cell carcinomas: interaction of tumour cells and lung parenchyma in the tumor periphery. Int. J. Cancer 37:11–19; 1986.

    Article  PubMed  CAS  Google Scholar 

  10. Easty, D. M.; Easty, G. C. Measurement of the ability of cells to infiltrate normal tissues in vitro. Br. J. Cancer 29:36–49; 1974.

    PubMed  CAS  Google Scholar 

  11. Engebraaten, O.; Bjerkvig, R.; Berens, M. E. Effect of alkyl-lysophospholipid on glioblastoma cell invasion into fetal rat brain tissue in vitro. Cancer Res. 51:1713–1719; 1991.

    PubMed  CAS  Google Scholar 

  12. Engebraaten, O.; Bjerkvig, R.; Lund-Johansen, M., et al. Interaction between human brain tumour biopsies and fetal rat brain tissue in vitro. Acta Neuropathol. 81:130–140; 1990.

    Article  PubMed  CAS  Google Scholar 

  13. Fidler, I. J. Rationale and methods for the use of nude mice to study the biology and therapy of human cancer metastasis. Cancer Metastasis Rev. 5:29–49; 1986.

    Article  PubMed  CAS  Google Scholar 

  14. Fjellbirkeland, L.; Bjerkvig, R.; Laerum, O. D. Tumour fragment spheroids from human non-small-cell lung cancer maintained in organ culture. Virchows Arch. 426:169–178; 1995.

    Article  PubMed  CAS  Google Scholar 

  15. Fjellbirkeland, L.; Bjerkvig, R.; Steinsvåg, S. K., et al. Nonadhesive stationary organ culture of human bronchial mucosa. Am. J. Respir. Cell Mol. Biol. 15:197–206; 1996.

    PubMed  CAS  Google Scholar 

  16. Fligiel, S. E. G.; Varani, J. In situ epithelial cell invasion in organ culture. Invasion & Metastasis 13:225–233; 1993.

    CAS  Google Scholar 

  17. Freshney, R. I. Culture of animal cells: a manual of basic techniques. 3rd ed. New York: Wiley-Liss; 1994.

    Google Scholar 

  18. Gazdar, A. F. The molecular and cellular basis of human lung cancer. Anticancer Res. 13:261–268; 1994.

    Google Scholar 

  19. Grümmer, R.; Hohn, H.-P.; Mareel, M. M., et al. Adhesion and invasion of three human choriocarcinoma cell lines into human endometrium in a three dimensional organ culture system. Placenta 15:411–429; 1994.

    PubMed  Google Scholar 

  20. Hiddemann, W.; Schumann, J.; Andreeff, M., et al. Convention on nomenclature for DNA cytometry. Cytometry 5:445–446; 1984.

    Article  Google Scholar 

  21. Honig, M. G.; Hume, R. I. Fluorescent carbocyanine dyes allow living neurons of identified origin to be studied in long-term cultures. J. Cell Biol. 103:171–187; 1986.

    Article  PubMed  CAS  Google Scholar 

  22. Hsu, S.-M.; Raine, L.; Fanger, H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J. Histochem. Cytochem. 29:577–580; 1981.

    PubMed  CAS  Google Scholar 

  23. Kluck, R. M.; McDougall, C. A.; Harmon, B. V., et al. Calcium chelators induce apoptosis—evidence that raised intracellular ionised calcium is not essential for apoptosis. Biochim. Biophys. Acta 1223:247–254; 1994.

    Article  PubMed  CAS  Google Scholar 

  24. Kohn, E. C.; Liotta, L. A. Molecular insights into cancer invasion: strategies for prevention and intervention. Cancer Res. 55:1856–1862; 1995.

    PubMed  CAS  Google Scholar 

  25. Liotta, L. A.; Kohn, E. C. Invasion and metastasis. In: Pine, J. W., Jr., ed. Cancer medicine. Vol. 1. 4th ed. Baltimore: Williams & Wilkins; 1997:165–180.

    Google Scholar 

  26. Liotta, L. A.; Lee, C. W.; Morakis, D. J. New method for preparing large surface of intact human basement membrane for tumor invasion studies. Cancer Lett. 11:141–152; 1980.

    Article  PubMed  CAS  Google Scholar 

  27. Liu, C.; Tsao, M.-S. Proto-oncogene and growth factor/receptor expression in the establishment of primary human non-small cell lung carcinoma cell lines. Am. J. Pathol. 142:413–423; 1993.

    PubMed  CAS  Google Scholar 

  28. Mareel, M. M.; Van Roy, F. M.; Bracke, M. E. How and when do tumor cells metastasize? Crit. Rev. Oncog. 4:559–594; 1993.

    PubMed  CAS  Google Scholar 

  29. Minna, J. D. Tumor suppressor genes and oncogenes in lung cancer: potential clinical applications. Adv. Oncol. 12:3–7; 1996.

    Google Scholar 

  30. Momiki, S.; Baba, M.; Caamano, J., et al. In vivo and in vitro invasiveness of human lung carcinoma cell lines. Invasion & Metastasis 11:66–75; 1991.

    CAS  Google Scholar 

  31. Mountain, C. F. Value of the new TNM staging system for lung cancer. Chest (Suppl.) 96:47S-49S; 1989.

    CAS  Google Scholar 

  32. Noel, A. C.; Callé, A.; Emonard, H. P., et al. Invasion of reconstituted basement membrane matrix is not correlated to the malignant metastatic cell phenotype. Cancer Res. 51:405–414; 1991.

    PubMed  CAS  Google Scholar 

  33. Simon, N.; Noel, A.; Foidart, J.-M. Evaluation of in vitro reconstituted basement membrane assay to assess the invasiveness of tumor cells. Invasion & Metastasis 12:156–167; 1992.

    CAS  Google Scholar 

  34. Starkey, J. R.; Hosick, H. L.; Stanford, D. R., et al. Interaction of metastatic tumor cells with bovine lens capsule basement membrane. Cancer Res. 44:1585–1594; 1984.

    PubMed  CAS  Google Scholar 

  35. Terranova, V. P.; Hujanen, E. S.; Loeb, D. M., et al. Use of reconstituted basement membrane to measure cell invasiveness and select for highly invasive tumor cells. Proc. Natl. Acad. Sci. USA 83:465–469; 1986.

    Article  PubMed  CAS  Google Scholar 

  36. Trump, B. F.; Harris, C. C. Human tissues in biomedical research. Hum. Pathol. 10:245–248; 1979.

    Article  PubMed  CAS  Google Scholar 

  37. Tucker, G. C.; Delouvee, A.; Jouanneau, J., et al. Amplification of invasiveness in organotypic cultures after NBT-II rat bladder carcinoma stimulation with in vitro scattering factors. Invasion & Metastasis 11:297–309; 1991.

    CAS  Google Scholar 

  38. Uphoff, C. C.; Gignac, S. M.; Drexler, H. G. Mycoplasma contamination in human leukemia cell lines I. Comparison of various detection methods. J. Immunol. Methods 149:43–53; 1992.

    PubMed  CAS  Google Scholar 

  39. Wyllie, A. H. Apoptosis and the regulation of cell numbers in normal and neoplastic tissues: an overview. Cancer Metastasis Rev. 11:95–103; 1992.

    Article  PubMed  CAS  Google Scholar 

  40. Yuhas, J. M.; Li, A. P.; Martinez, A. O., et al. A simplified method for production and growth of multicellular tumor spheroids. Cancer Res. 37:3639–3643; 1977.

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Pathology, The Gade Institute, University of Bergen, N-5021, Bergen, Norway

    Lars Fjellbirkeland & Ole Didrik Laerum

  2. Department of Thoracic Medicine, University of Bergen, N-5021, Bergen, Norway

    Lars Fjellbirkeland

  3. Department of Anatomy and Cell Biology, University of Bergen, N-5009, Bergen, Norway

    Rolf Bjerkvig

Authors
  1. Lars Fjellbirkeland
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Rolf Bjerkvig
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ole Didrik Laerum
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Fjellbirkeland, L., Bjerkvig, R. & Laerum, O.D. Non-small-cell lung carcinoma cells invade human bronchial mucosa In vitro . In Vitro Cell.Dev.Biol.-Animal 34, 333–340 (1998). https://doi.org/10.1007/s11626-998-0010-4

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-998-0010-4

Key words

Search

Navigation