Abstract
Purpose
Prostate cancer (PCa) becomes lethal when cancer cells develop into castration-resistant PCa, which remains incurable because of the poor understanding of their cell origin and characteristics. We aim to investigate the potential role of cancer stem cells (CSCs) in PCa progression.
Methods
Human PCa cell lines (LNCaP, 22RV1, DU145 and PC-3) were plated in serum-free suspension culture system allowed for tumorsphere forming. To evaluate the CSC characteristics of tumorspheres, the self-renewal, chemoresistance, tumorigenicity of the PCa tumorsphere cells, and the expression levels of stemness-related proteins in the PCa tumorsphere cells were assessed, comparing with the parental adherent cells.
Results
Tumorsphere cells from PCa cell lines displayed enhanced self-renewal, chemoresistance and tumor-initiating capacity when compared with the adherent cells. Additionally, these cells overexpressed CSC marker CD44. Also, the tumorsphere cells expressed high levels of “stemness” genes Gli1, ABCG2 and Bmi-1.
Conclusions
Collectively, these data demonstrated that tumorspheres derived from PCa cells possess chemoresistant and CSC properties. Our study suggests that the identification of PCa CSCs could provide new insight into the lethal phenotype of PCa and therapeutic implications.
Similar content being viewed by others
References
Ao A, Morrison BJ, Wang H, Lopez JA, Reynolds BA, Lu J (2011) Response of estrogen receptor-positive breast cancer tumorspheres to antiestrogen treatments. PLoS One 6(4):e18810. doi:10.1371/journal.pone.0018810
Balic M, Lin H, Young L, Hawes D, Giuliano A, McNamara G, Datar RH, Cote RJ (2006) Most early disseminated cancer cells detected in bone marrow of breast cancer patients have a putative breast cancer stem cell phenotype. Clin Cancer Res 12(19):5615–5621. doi:10.1158/1078-0432.CCR-06-0169
Bapat SA, Mali AM, Koppikar CB, Kurrey NK (2005) Stem and progenitor-like cells contribute to the aggressive behavior of human epithelial ovarian cancer. Cancer Res 65(8):3025–3029. doi:10.1158/0008-5472.CAN-04-3931
Bisson I, Prowse DM (2009) WNT signaling regulates self-renewal and differentiation of prostate cancer cells with stem cell characteristics. Cell Res 19(6):683–697. doi:10.1038/cr.2009.43
Bruggeman SW, Hulsman D, Tanger E, Buckle T, Blom M, Zevenhoven J, van Tellingen O, van Lohuizen M (2007) Bmi1 controls tumor development in an Ink4a/Arf-independent manner in a mouse model for glioma. Cancer Cell 12(4):328–341. doi:10.1016/j.ccr.2007.08.032
Cao L, Bombard J, Cintron K, Sheedy J, Weetall ML, Davis TW (2011a) BMI1 as a novel target for drug discovery in cancer. J Cell Biochem 112(10):2729–2741. doi:10.1002/jcb.23234
Cao L, Zhou Y, Zhai B, Liao J, Xu W, Zhang R, Li J, Zhang Y, Chen L, Qian H, Wu M, Yin Z (2011b) Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines. BMC Gastroenterol 11:71. doi:10.1186/1471-230X-11-71
Ding XW, Wu JH, Jiang CP (2010) ABCG2: a potential marker of stem cells and novel target in stem cell and cancer therapy. Life Sci 86(17–18):631–637. doi:10.1016/j.lfs.2010.02.012
Fang D, Nguyen TK, Leishear K, Finko R, Kulp AN, Hotz S, Van Belle PA, Xu X, Elder DE, Herlyn M (2005) A tumorigenic subpopulation with stem cell properties in melanomas. Cancer Res 65(20):9328–9337. doi:10.1158/0008-5472.CAN-05-1343
Feldman BJ, Feldman D (2001) The development of androgen-independent prostate cancer. Nat Rev Cancer 1(1):34–45. doi:10.1038/35094009
Fujii H, Honoki K, Tsujiuchi T, Kido A, Yoshitani K, Takakura Y (2009) Sphere-forming stem-like cell populations with drug resistance in human sarcoma cell lines. Int J Oncol 34(5):1381–1386
Ghods AJ, Irvin D, Liu G, Yuan X, Abdulkadir IR, Tunici P, Konda B, Wachsmann-Hogiu S, Black KL, Yu JS (2007) Spheres isolated from 9L gliosarcoma rat cell line possess chemoresistant and aggressive cancer stem-like cells. Stem Cells 25(7):1645–1653. doi:10.1634/stemcells.2006-0624
Gilbert CA, Ross AH (2009) Cancer stem cells: cell culture, markers, and targets for new therapies. J Cell Biochem 108(5):1031–1038. doi:10.1002/jcb.22350
Gulino A, Ferretti E, De Smaele E (2009) Hedgehog signalling in colon cancer and stem cells. EMBO Mol Med 1(6–7):300–302. doi:10.1002/emmm.200900042
Hong SP, Wen J, Bang S, Park S, Song SY (2009) CD44-positive cells are responsible for gemcitabine resistance in pancreatic cancer cells. Int J Cancer 125(10):2323–2331. doi:10.1002/ijc.24573
Jemal A, Siegel R, Xu J, Ward E (2010) Cancer statistics 2010. CA Cancer J Clin 60(5):277–300. doi:10.3322/caac.20073
Kelly K, Yin JJ (2008) Prostate cancer and metastasis initiating stem cells. Cell Res 18(5):528–537. doi:10.1038/cr.2008.50
Kim M, Turnquist H, Jackson J, Sgagias M, Yan Y, Gong M, Dean M, Sharp JG, Cowan K (2002) The multidrug resistance transporter ABCG2 (breast cancer resistance protein 1) effluxes Hoechst 33342 and is overexpressed in hematopoietic stem cells. Clin Cancer Res 8(1):22–28
Kondo T, Setoguchi T, Taga T (2004) Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line. Proc Natl Acad Sci USA 101(3):781–786. doi:10.1073/pnas.0307618100
Lang SH, Frame FM, Collins AT (2009) Prostate cancer stem cells. J Pathol 217(2):299–306. doi:10.1002/path.2478
Li C, Heidt DG, Dalerba P, Burant CF, Zhang L, Adsay V, Wicha M, Clarke MF, Simeone DM (2007) Identification of pancreatic cancer stem cells. Cancer Res 67(3):1030–1037. doi:10.1158/0008-5472.CAN-06-2030
Li H, Jiang M, Honorio S, Patrawala L, Jeter CR, Calhoun-Davis T, Hayward SW, Tang DG (2009) Methodologies in assaying prostate cancer stem cells. Methods Mol Biol 568:85–138. doi:10.1007/978-1-59745-280-9_7
Liu T, Xu F, Du X, Lai D, Zhao Y, Huang Q, Jiang L, Huang W, Cheng W, Liu Z (2010) Establishment and characterization of multi-drug resistant, prostate carcinoma-initiating stem-like cells from human prostate cancer cell lines 22RV1. Mol Cell Biochem 340(1–2):265–273. doi:10.1007/s11010-010-0426-5
Liu C, Kelnar K, Liu B, Chen X, Calhoun-Davis T, Li H, Patrawala L, Yan H, Jeter C, Honorio S, Wiggins JF, Bader AG, Fagin R, Brown D, Tang DG (2011) The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44. Nat Med 17(2):211–215. doi:10.1038/nm.2284
Maitland NJ, Collins AT (2008) Prostate cancer stem cells: a new target for therapy. J Clin Oncol 26(17):2862–2870. doi:10.1200/JCO.2007.15.1472
Marhaba R, Klingbeil P, Nuebel T, Nazarenko I, Buechler MW, Zoeller M (2008) CD44 and EpCAM: cancer-initiating cell markers. Curr Mol Med 8(8):784–804
Marshall GP 2nd, Reynolds BA, Laywell ED (2007) Using the neurosphere assay to quantify neural stem cells in vivo. Curr Pharm Biotechnol 8(3):141–145
O’Brien CA, Pollett A, Gallinger S, Dick JE (2007) A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature 445(7123):106–110. doi:10.1038/nature05372
Olsson E, Honeth G, Bendahl PO, Saal LH, Gruvberger-Saal S, Ringner M, Vallon-Christersson J, Jonsson G, Holm K, Lovgren K, Ferno M, Grabau D, Borg A, Hegardt C (2011) CD44 isoforms are heterogeneously expressed in breast cancer and correlate with tumor subtypes and cancer stem cell markers. BMC Cancer 11(1):418. doi:10.1186/1471-2407-11-418
Orian-Rousseau V (2010) CD44, a therapeutic target for metastasising tumours. Eur J Cancer 46(7):1271–1277. doi:10.1016/j.ejca.2010.02.024
Patrawala L, Calhoun T, Schneider-Broussard R, Li H, Bhatia B, Tang S, Reilly JG, Chandra D, Zhou J, Claypool K, Coghlan L, Tang DG (2006) Highly purified CD44+ prostate cancer cells from xenograft human tumors are enriched in tumorigenic and metastatic progenitor cells. Oncogene 25(12):1696–1708. doi:10.1038/sj.onc.1209327
Pfeiffer MJ, Schalken JA (2010) Stem cell characteristics in prostate cancer cell lines. Eur Urol 57(2):246–254. doi:10.1016/j.eururo.2009.01.015
Pienta KJ, Bradley D (2006) Mechanisms underlying the development of androgen-independent prostate cancer. Clin Cancer Res 12(6):1665–1671. doi:10.1158/1078-0432.CCR-06-0067
Ponti D, Costa A, Zaffaroni N, Pratesi G, Petrangolini G, Coradini D, Pilotti S, Pierotti MA, Daidone MG (2005) Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res 65(13):5506–5511. doi:10.1158/0008-5472.CAN-05-0626
Rappa G, Mercapide J, Anzanello F, Prasmickaite L, Xi Y, Ju J, Fodstad O, Lorico A (2008) Growth of cancer cell lines under stem cell-like conditions has the potential to unveil therapeutic targets. Exp Cell Res 314(10):2110–2122. doi:10.1016/j.yexcr.2008.03.008
Rich JN, Bao S (2007) Chemotherapy and cancer stem cells. Cell Stem Cell 1(4):353–355. doi:10.1016/j.stem.2007.09.011
Ruiz i Altaba A, Mas C, Stecca B (2007) The Gli code: an information nexus regulating cell fate, stemness and cancer. Trends Cell Biol 17(9):438–447. doi:10.1016/j.tcb.2007.06.007
Sharom FJ (2008) ABC multidrug transporters: structure, function and role in chemoresistance. Pharmacogenomics 9(1):105–127. doi:10.2217/14622416.9.1.105
Singh SK, Clarke ID, Terasaki M, Bonn VE, Hawkins C, Squire J, Dirks PB (2003) Identification of a cancer stem cell in human brain tumors. Cancer Res 63(18):5821–5828
Singh SK, Hawkins C, Clarke ID, Squire JA, Bayani J, Hide T, Henkelman RM, Cusimano MD, Dirks PB (2004) Identification of human brain tumour initiating cells. Nature 432(7015):396–401. doi:10.1038/nature03128
Song Z, Yue W, Wei B, Wang N, Li T, Guan L, Shi S, Zeng Q, Pei X, Chen L (2011) Sonic hedgehog pathway is essential for maintenance of cancer stem-like cells in human gastric cancer. PLoS One 6(3):e17687. doi:10.1371/journal.pone.0017687
Su YJ, Lai HM, Chang YW, Chen GY, Lee JL (2011) Direct reprogramming of stem cell properties in colon cancer cells by CD44. EMBO J 30(15):3186–3199. doi:10.1038/emboj.2011.211emboj2011211
Takebe N, Harris PJ, Warren RQ, Ivy SP (2011) Targeting cancer stem cells by inhibiting Wnt, Notch, and Hedgehog pathways. Nat Rev Clin Oncol 8(2):97–106. doi:10.1038/nrclinonc.2010.196
Visvader JE, Lindeman GJ (2008) Cancer stem cells in solid tumours: accumulating evidence and unresolved questions. Nat Rev Cancer 8(10):755–768. doi:10.1038/nrc2499
Wang X, Venugopal C, Manoranjan B, McFarlane N, O’Farrell E, Nolte S, Gunnarsson T, Hollenberg R, Kwiecien J, Northcott P, Taylor MD, Hawkins C, Singh SK (2011) Sonic hedgehog regulates Bmi1 in human medulloblastoma brain tumor-initiating cells. Oncogene. doi:10.1038/onc.2011.232onc2011232
Wicha MS (2006) Cancer stem cells and metastasis: lethal seeds. Clin Cancer Res 12(19):5606–5607. doi:10.1158/1078-0432.CCR-06-1537
Yu CC, Lo WL, Chen YW, Huang PI, Hsu HS, Tseng LM, Hung SC, Kao SY, Chang CJ, Chiou SH (2011) Bmi-1 regulates snail expression and promotes metastasis ability in head and neck squamous cancer-derived ALDH1 positive cells. J Oncol 2011, art no 609259. doi:10.1155/2011/609259
Zen Y, Fujii T, Yoshikawa S, Takamura H, Tani T, Ohta T, Nakanuma Y (2007) Histological and culture studies with respect to ABCG2 expression support the existence of a cancer cell hierarchy in human hepatocellular carcinoma. Am J Pathol 170(5):1750–1762. doi:10.2353/ajpath.2007.060798
Zhang Y, Li B, Ji ZZ, Zheng PS (2010) Notch1 regulates the growth of human colon cancers. Cancer 116(22):5207–5218. doi:10.1002/cncr.25449
Zhao C, Chen A, Jamieson CH, Fereshteh M, Abrahamsson A, Blum J, Kwon HY, Kim J, Chute JP, Rizzieri D, Munchhof M, VanArsdale T, Beachy PA, Reya T (2009) Hedgehog signalling is essential for maintenance of cancer stem cells in myeloid leukaemia. Nature 458(7239):776–779. doi:10.1038/nature07737
Zheng X, Shen G, Yang X, Liu W (2007) Most C6 cells are cancer stem cells: evidence from clonal and population analyses. Cancer Res 67(8):3691–3697. doi:10.1158/0008-5472.CAN-06-3912
Zhong Y, Guan K, Guo S, Zhou C, Wang D, Ma W, Zhang Y, Li C, Zhang S (2010) Spheres derived from the human SK-RC-42 renal cell carcinoma cell line are enriched in cancer stem cells. Cancer Lett 299(2):150–160. doi:10.1016/j.canlet.2010.08.013
Acknowledgments
This study was supported by a grant from National Natural Science Foundation of China (No. 30801152 to LL Zhang).
Conflict of interest
We declare that we have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Zhang, L., Jiao, M., Li, L. et al. Tumorspheres derived from prostate cancer cells possess chemoresistant and cancer stem cell properties. J Cancer Res Clin Oncol 138, 675–686 (2012). https://doi.org/10.1007/s00432-011-1146-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-011-1146-2