Morpho-physiological heterogeneity of cells within two rat prostate carcinoma cell lines AT-2 and MAT-LyLu differing in the degree of malignancy observed by cell cloning and the effects of caffeine, theophylline and papaverine upon a proportion of the clones

Musialik,Ewa; Ryszawy,Damian; Madeja,Zbigniew; Korohoda,Wlodzimierz

doi:10.3892/or.2013.2323

Morpho-physiological heterogeneity of cells within two rat prostate carcinoma cell lines AT-2 and MAT-LyLu differing in the degree of malignancy observed by cell cloning and the effects of caffeine, theophylline and papaverine upon a proportion of the clones

Authors:
- Ewa Musialik
- Damian Ryszawy
- Zbigniew Madeja
- Wlodzimierz Korohoda
View Affiliations

Affiliations: Department of Cell Biology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, 30-387 Krakow, Poland
Published online on: March 5, 2013 https://doi.org/10.3892/or.2013.2323
Pages: 1789-1796

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Analysis of the heterogeneity of clones of single cells from two established lines of the Dunning series of rat prostate carcinoma differing in the degree of malignancy, AT-2 (moderately malignant) and MAT-LyLu (highly malignant), was conducted. The results showed that not only the original tumors and primary cell cultures were heterogeneous but also the established cell lines of tumor origin. In the MAT-LyLu cell line, the clones of cells with morpho-physiological features characteristic of malignant cells dominated, whereas diverse types of clones were present in the AT-2 cell line. Differences in cell morphology (EMT), multi-layering and release from contact inhibition followed by active migration were observed and found to be correlated with increased expression of proteins involved in cancer cell invasiveness: connexin 43 and transcription factor Snail. Caffeine, theophylline and papaverine, reported to show anticancer activity in vivo, were found to decrease the proportion of clones displaying malignant cell features in the AT-2 cell line. At the tested concentrations, these compounds reversibly retarded cell growth but did not inhibit it. The results showed that the heterogeneity of cell populations within the cell lines should be taken into account in experiments carried out in vitro on established model cancer cell lines.

View Figures

View References

1	Wang N, Wilkin A, Böcking A and Tribukait B: Evaluation of tumor heterogeneity of prostate carcinoma by flow and image DNA cytometry and histopathological grading. Anal Cell Physiol. 20:49–62. 2000.PubMed/NCBI
2	Singh RK and Talmadge JE: The evolution of diversity within tumors and metastases. Selected Aspects of Cancer Progression: Metastasis, Apoptosis and Immune Response. Kaiser HE and Nasir A: Springer Science-Business Media BV; Dordrecht: pp. 59–90. 2008
3	Marusyk A and Polyak K: Tumor heterogeneity: causes and consequences. Biochim Biophys Acta. 1805:105–117. 2010.PubMed/NCBI
4	Dirks P: Cancer stem cells. Invitation to a second round. Nature. 466:40–41. 2010. View Article : Google Scholar : PubMed/NCBI
5	Carter HB and Coffey DS: Cell surface charge in predicting metastatic potential of aspirated cells from the Dunning rat prostate adenocarcinoma model. J Urol. 140:173–175. 1988.PubMed/NCBI
6	Carter HB, Partin AW and Coffey DS: Cell surface charge in predicting metastatic potential in an animal model of prostate cancer: flow cytometric quantification of cell surface charge. J Urol. 142:1338–1341. 1989.PubMed/NCBI
7	Fraser SP, Ding Y, Liu A, Foster CS and Djamgoz MBA: Tetrodotoxin suppress morphological enhancement of the metastatic MAT-LyLu rat prostate cell line. Cell Tissue Res. 295:505–512. 1999. View Article : Google Scholar : PubMed/NCBI
8	Bironaite D, Nesland JM, Dalen H, Risberg B and Bryne M: N-Glycans influence the in vitro adhesive and invasive behaviour of three metastatic cell lines. Tumor Biol. 200:165–175. 2000. View Article : Google Scholar : PubMed/NCBI
9	Djamgoz MBA, Mycielska M, Madeja Z, Fraser SP and Korohoda W: Directional movement of rat prostate cancer cells in direct-current electric field. Involvement of voltage gated Na⁺ channel activity J Cell Sci. 114:2697–2705. 2001.PubMed/NCBI
10	Pokorna E, Zicha D, Chaloupkova A, Matousková E and Vesely P: Two dynamic morphotypes of sarcoma cells, asymmetric stellate and triangle with leading lamella, are related to malignancy. Folia Biol. 49:33–39. 2003.PubMed/NCBI
11	Sanford KK, Likely GD and Earle WR: The development of variations in transplantability and morphology within a clone of mouse fibroblasts transformed to sarcoma-producing cells in vitro. J Natl Cancer Inst. 15:215–237. 1954.PubMed/NCBI
12	Abercrombie M and Ambrose EJ: The surface properties of cancer cells: a review. Cancer Res. 22:332–345. 1962.
13	Moscona A, Trowell QA and Willmer EN: Methods. (Chap. 2). Cell and Tissues in Culture. 1. Willmer EN: Academic Press; London: pp. 1–98. 1965
14	Burger MM: The significance of surface structure changes for growth control under crowded conditions. Growth Control in Cell Cultures. Wolstenhome GEW and Knight J: Churchill Livingstone; London: pp. 45–62. 1971
15	Wyckoff JB, Segall JE and Condeelis JS: The collection of the motile population of cells from a living tumor. Cancer Res. 60:5401–5404. 2000.PubMed/NCBI
16	Thiery JP: Epithelial-mesenchymal transitions in development and pathogenesis. Curr Opin Cell Biol. 15:740–746. 2003. View Article : Google Scholar : PubMed/NCBI
17	Savagner P: The epithelial-mesenchymal transition (EMT) phenomenon. Ann Oncol. 21(Suppl 7): vii89–vii92. 2010. View Article : Google Scholar : PubMed/NCBI
18	Graham TR, Zhau H, Odera-Marah VA, Osunkoya AO, Kimbrok S, Tighiouart M, Liu T, Simons JW and O'Regan RM: Insulin-like growth factor-I-dependent up-regulation of ZEB1 drives epithelial-to-mesenchymal transition in human prostate cancer cells. Cancer Res. 68:2479–2488. 2008. View Article : Google Scholar : PubMed/NCBI
19	Klarmann GJ, Hurt EM, Matheus LA, Zhang X, Duhagon MA, Mistree T, Thomas SB and Farrar WL: Invasive prostate cancer cells are tumor initiating cells that have a stem cell-like genomic signature. Clin Exp Metastasis. 26:433–446. 2009. View Article : Google Scholar : PubMed/NCBI
20	Tate AW, Lung T, Radhakrishnan A, Lim SD, Lin X and Edlund M: Changes in gap junctional connexin isoforms during prostate cancer progression. Prostate. 66:19–31. 2006. View Article : Google Scholar : PubMed/NCBI
21	Kanczuga-Koda L, Sulkowski S, Lenczewski A, Koda M, Wincewicz A, Baltaziak M and Sulkowska M: Increased expression of connexins 26 and 43 in lymph node metastases of breast cancer. J Clin Pathol. 59:429–433. 2006. View Article : Google Scholar : PubMed/NCBI
22	Baritaki S, Chapman A, Yeung K, Spandidos DA, Palladino M and Bonavida B: Inhibition of epithelial to mesenchymal transition in metastatic prostate cancer cells by the novel proteasome inhibitor, NPI-0052: pivotal roles of Snail repression and RKIP induction. Oncogene. 28:3573–3585. 2009. View Article : Google Scholar
23	Czyz J, Szpak K and Madeja Z: The role of connexins in prostate cancer promotion and progression. Nat Rev Urol. 9:274–282. 2012. View Article : Google Scholar : PubMed/NCBI
24	Lentini A, Mattioli P, Nicolini L, Pietrini A, Abbruzzese A and Beninati S: Anti-invasive effects of theophylline on experimental B16–F10 melanoma lung metastasis. Cancer J. 10:274–278. 1997.PubMed/NCBI
25	Spangler JG: Bone biology and physiology: implications for novel osteoblastic osteosarcoma treatments. Med Hypotheses. 70:281–286. 2008. View Article : Google Scholar : PubMed/NCBI
26	Holick CN, Smith SG, Giovannucci E and Michaud DS: Coffee, tea, caffeine intake, and risk of adult glioma in three prospective cohort studies. Cancer Epidemiol Biomarkers Prev. 19:39–47. 2010. View Article : Google Scholar : PubMed/NCBI
27	Wilson KM, Kasperzyk JL, Rider JR, Kenfield S, Van Dam RM, Stampfer MJ, Giovannucci E and Mucci LA: Coffee consumption and prostate risk and progression in the health professionals follow up study. J Natl Cancer Inst. 103:876–884. 2011. View Article : Google Scholar
28	Isaacs JT: The R-3327 system of rat prostatic cancers. Urol Oncol. 2:115–116. 1996. View Article : Google Scholar : PubMed/NCBI
29	Ramaekers FC, Verhagen AP, Isaacs JT, Feitz WF, Moesker O, Schaart G, Schalken JA and Voijs GP: Intermediate filament expression and the progression of prostatic cancer as studied in the Dunning R-3327 rat prostatic carcinoma system. Prostate. 14:323–339. 1989. View Article : Google Scholar : PubMed/NCBI
30	Erickson CA and Nuccitelli R: Embryonic fibroblast motility and orientation can be influenced by physiological electric fields. J Cell Biol. 98:296–307. 1984. View Article : Google Scholar : PubMed/NCBI
31	Korohoda W and Madeja Z: Contact of sarcoma cells with aligned fibroblasts accelerated their displacement: computer-assisted analysis of tumour cell locomotion in co-culture. Biochem Cell Biol. 75:263–276. 1997. View Article : Google Scholar
32	Waligorska A, Wianecka-Skoczen M, Nowak P and Korohoda W: Some difficulties in research into cell motile activity under isotropic conditions. Folia Biol. 55:9–16. 2007. View Article : Google Scholar : PubMed/NCBI
33	Daniel-Wojcik A, Misztal K, Bechyne I, Sroka J, Miekus K, Madeja Z and Czyz J: Cell motility affects the intensity of gap junctional coupling in prostate carcinoma and melanoma cell populations. Int J Oncol. 33:309–315. 2009.PubMed/NCBI
34	Miekus K, Czernik M, Sroka J, Czyz J and Madeja Z: Contact stimulation of prostate cancer cell migration: the role of gap junctional coupling and migration stimulated by heterotypic cell-to-cell contacts in determination of the metastatic phenotype of Dunning rat prostate cancer cells. Biol Cell. 97:893–903. 2005. View Article : Google Scholar
35	Burke JM, Skumasz CMB, Irwing PE and McKay BS: Phenotypic heterogeneity of retinoid pigment epithelial cells in vitro and in situ. Exp Eye Res. 62:63–73. 1996. View Article : Google Scholar : PubMed/NCBI
36	Forrester JA: Microelectrophoresis of normal and polyoma virus transformed hamster kidney fibroblasts. Cell Electrophoresis. Ambrose EJ: J. & A. Churchill Ltd; London: pp. 115–124. 1965
37	Korohoda W and Czyz J: Efficacy of the Frame and Hu mathematical model for the quantitative analysis of agents influencing growth of chick embryo fibroblasts. Folia Histochem Cytobiol. 32:113–118. 1994.PubMed/NCBI
38	Prasad KN and Sheppard JR: Inhibitor of cyclic-nucleotide phosphodiesterase tumor cells induce morphological differentiation of mouse neuroblastoma cell culture. Exp Cell Res. 73:436–440. 1972. View Article : Google Scholar
39	Weiss L and Subjeck JR: Electrical heterogeneity of the surfaces of Ehrlich ascites tumor cells. Ann NY Acad Sci. 238:352–361. 1974. View Article : Google Scholar : PubMed/NCBI