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. 2001 Aug;85(4):590–599. doi: 10.1054/bjoc.2001.1967

Prostate epithelial cell lines form spheroids with evidence of glandular differentiation in three-dimensional Matrigel cultures

S H Lang 1, R M Sharrard 1, M Stark 2, J M Villette 3, N J Maitland 1
PMCID: PMC2364090  PMID: 11506501

Abstract

Normal (PNT2-C2) and metastatic (PC-3) prostate cell lines were grown in Matrigel to observe the effects on morphology and phenotype in comparison to monolayer culture. In monolayer cultures, PNT2-C2 showed typical round/cuboidal epithelial morphology, with tight cell associations, whereas in Matrigel they formed smooth spheroids, tightly packed with cells. In both monolayer and Matrigel, PNT2-C2 had a differentiated luminal epithelial phenotype with high expression of cytokeratin 8, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), E-cadherin and desmoglein. In contrast, PC-3 cells possessed an epithelial/mesenchyme morphology in monolayer with loose cell to cell contact and pseudopodial extensions. Immunohistochemical phenotyping indicated the cells were undifferentiated, expressing high levels of vimentin, β1 integrin, CD44 and low expression of cytokeratin 8. In Matrigel they formed smooth and irregular spheroids, which had a lumen surrounded by a single cell layer. Matrigel also influenced the expression of PSA, PSMA and CD44. These results indicate that Matrigel culture can induce morphological differentiation of prostate cancer cells which initially had a basal phenotype. © 2001 CancerResearch Campaign http://www.bjcancer.com

Keywords: Matrigel, differentiation, prostate spheroids

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Selected References

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  1. Bae S. N., Arand G., Azzam H., Pavasant P., Torri J., Frandsen T. L., Thompson E. W. Molecular and cellular analysis of basement membrane invasion by human breast cancer cells in Matrigel-based in vitro assays. Breast Cancer Res Treat. 1993;24(3):241–255. doi: 10.1007/BF01833264. [DOI] [PubMed] [Google Scholar]
  2. Barcellos-Hoff M. H., Aggeler J., Ram T. G., Bissell M. J. Functional differentiation and alveolar morphogenesis of primary mammary cultures on reconstituted basement membrane. Development. 1989 Feb;105(2):223–235. doi: 10.1242/dev.105.2.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bayne C. W., Donnelly F., Chapman K., Bollina P., Buck C., Habib F. A novel coculture model for benign prostatic hyperplasia expressing both isoforms of 5 alpha-reductase. J Clin Endocrinol Metab. 1998 Jan;83(1):206–213. doi: 10.1210/jcem.83.1.4486. [DOI] [PubMed] [Google Scholar]
  4. Bouwens L., De Blay E. Islet morphogenesis and stem cell markers in rat pancreas. J Histochem Cytochem. 1996 Sep;44(9):947–951. doi: 10.1177/44.9.8773559. [DOI] [PubMed] [Google Scholar]
  5. Chang S. S., Reuter V. E., Heston W. D., Bander N. H., Grauer L. S., Gaudin P. B. Five different anti-prostate-specific membrane antigen (PSMA) antibodies confirm PSMA expression in tumor-associated neovasculature. Cancer Res. 1999 Jul 1;59(13):3192–3198. [PubMed] [Google Scholar]
  6. Chen L. H., Bissell M. J. A novel regulatory mechanism for whey acidic protein gene expression. Cell Regul. 1989 Nov;1(1):45–54. doi: 10.1091/mbc.1.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Cussenot O., Berthon P., Berger R., Mowszowicz I., Faille A., Hojman F., Teillac P., Le Duc A., Calvo F. Immortalization of human adult normal prostatic epithelial cells by liposomes containing large T-SV40 gene. J Urol. 1991 Sep;146(3):881–886. doi: 10.1016/s0022-5347(17)37953-3. [DOI] [PubMed] [Google Scholar]
  9. Fraga C. H., True L. D., Kirk D. Enhanced expression of the mesenchymal marker, vimentin, in hyperplastic versus normal human prostatic epithelium. J Urol. 1998 Jan;159(1):270–274. doi: 10.1016/s0022-5347(01)64080-1. [DOI] [PubMed] [Google Scholar]
  10. Knox J. D., Cress A. E., Clark V., Manriquez L., Affinito K. S., Dalkin B. L., Nagle R. B. Differential expression of extracellular matrix molecules and the alpha 6-integrins in the normal and neoplastic prostate. Am J Pathol. 1994 Jul;145(1):167–174. [PMC free article] [PubMed] [Google Scholar]
  11. Lang S. H., Clarke N. W., George N. J., Testa N. G. Scatter factor influences the formation of prostate epithelial cell colonies on bone marrow stroma in vitro. Clin Exp Metastasis. 1999 Jun;17(4):333–340. doi: 10.1023/a:1006696002497. [DOI] [PubMed] [Google Scholar]
  12. Lang S. H., Stower M., Maitland N. J. In vitro modelling of epithelial and stromal interactions in non-malignant and malignant prostates. Br J Cancer. 2000 Feb;82(4):990–997. doi: 10.1054/bjoc.1999.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Liu A. Y., True L. D., LaTray L., Nelson P. S., Ellis W. J., Vessella R. L., Lange P. H., Hood L., van den Engh G. Cell-cell interaction in prostate gene regulation and cytodifferentiation. Proc Natl Acad Sci U S A. 1997 Sep 30;94(20):10705–10710. doi: 10.1073/pnas.94.20.10705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mitchell S., Abel P., Ware M., Stamp G., Lalani E. Phenotypic and genotypic characterization of commonly used human prostatic cell lines. BJU Int. 2000 May;85(7):932–944. doi: 10.1046/j.1464-410x.2000.00606.x. [DOI] [PubMed] [Google Scholar]
  15. Morton R. A., Ewing C. M., Nagafuchi A., Tsukita S., Isaacs W. B. Reduction of E-cadherin levels and deletion of the alpha-catenin gene in human prostate cancer cells. Cancer Res. 1993 Aug 1;53(15):3585–3590. [PubMed] [Google Scholar]
  16. Murant S. J., Handley J., Stower M., Reid N., Cussenot O., Maitland N. J. Co-ordinated changes in expression of cell adhesion molecules in prostate cancer. Eur J Cancer. 1997 Feb;33(2):263–271. doi: 10.1016/s0959-8049(96)00418-2. [DOI] [PubMed] [Google Scholar]
  17. Nagle R. B., Hao J., Knox J. D., Dalkin B. L., Clark V., Cress A. E. Expression of hemidesmosomal and extracellular matrix proteins by normal and malignant human prostate tissue. Am J Pathol. 1995 Jun;146(6):1498–1507. [PMC free article] [PubMed] [Google Scholar]
  18. Paradis V., Eschwège P., Loric S., Dumas F., Ba N., Benoît G., Jardin A., Bedossa P. De novo expression of CD44 in prostate carcinoma is correlated with systemic dissemination of prostate cancer. J Clin Pathol. 1998 Nov;51(11):798–802. doi: 10.1136/jcp.51.11.798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Pisters L. L., Troncoso P., Zhau H. E., Li W., von Eschenbach A. C., Chung L. W. c-met proto-oncogene expression in benign and malignant human prostate tissues. J Urol. 1995 Jul;154(1):293–298. [PubMed] [Google Scholar]
  20. Robinson E. J., Neal D. E., Collins A. T. Basal cells are progenitors of luminal cells in primary cultures of differentiating human prostatic epithelium. Prostate. 1998 Nov 1;37(3):149–160. doi: 10.1002/(sici)1097-0045(19981101)37:3<149::aid-pros4>3.0.co;2-e. [DOI] [PubMed] [Google Scholar]
  21. Sherwood E. R., Theyer G., Steiner G., Berg L. A., Kozlowski J. M., Lee C. Differential expression of specific cytokeratin polypeptides in the basal and luminal epithelia of the human prostate. Prostate. 1991;18(4):303–314. doi: 10.1002/pros.2990180404. [DOI] [PubMed] [Google Scholar]
  22. Smits A., van Grieken D., Hartman M., Lendahl U., Funa K., Nistér M. Coexpression of platelet-derived growth factor alpha and beta receptors on medulloblastomas and other primitive neuroectodermal tumors is consistent with an immature stem cell and neuronal derivation. Lab Invest. 1996 Jan;74(1):188–198. [PubMed] [Google Scholar]
  23. Smola H., Stark H. J., Thiekötter G., Mirancea N., Krieg T., Fusenig N. E. Dynamics of basement membrane formation by keratinocyte-fibroblast interactions in organotypic skin culture. Exp Cell Res. 1998 Mar 15;239(2):399–410. doi: 10.1006/excr.1997.3910. [DOI] [PubMed] [Google Scholar]
  24. Sommers C. L., Byers S. W., Thompson E. W., Torri J. A., Gelmann E. P. Differentiation state and invasiveness of human breast cancer cell lines. Breast Cancer Res Treat. 1994;31(2-3):325–335. doi: 10.1007/BF00666165. [DOI] [PubMed] [Google Scholar]
  25. Streuli C. H., Bissell M. J. Expression of extracellular matrix components is regulated by substratum. J Cell Biol. 1990 Apr;110(4):1405–1415. doi: 10.1083/jcb.110.4.1405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Timms B. G., Lee C. W., Aumüller G., Seitz J. Instructive induction of prostate growth and differentiation by a defined urogenital sinus mesenchyme. Microsc Res Tech. 1995 Mar 1;30(4):319–332. doi: 10.1002/jemt.1070300407. [DOI] [PubMed] [Google Scholar]
  27. Weaver V. M., Fischer A. H., Peterson O. W., Bissell M. J. The importance of the microenvironment in breast cancer progression: recapitulation of mammary tumorigenesis using a unique human mammary epithelial cell model and a three-dimensional culture assay. Biochem Cell Biol. 1996;74(6):833–851. doi: 10.1139/o96-089. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Webber M. M., Bello D., Kleinman H. K., Hoffman M. P. Acinar differentiation by non-malignant immortalized human prostatic epithelial cells and its loss by malignant cells. Carcinogenesis. 1997 Jun;18(6):1225–1231. doi: 10.1093/carcin/18.6.1225. [DOI] [PubMed] [Google Scholar]

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