A comprehensive panel of three-dimensional models for studies of prostate cancer growth, invasion and drug responses.
Prostate epithelial cells from both normal and cancer tissues, grown in three-dimensional (3D) culture as spheroids, represent promising in vitro models for the study of normal and cancer-relevant patterns of epithelial differentiation. We have developed the most comprehensive panel of miniaturized prostate cell culture models in 3D to date (n = 29), including many non-transformed and most currently available classic prostate cancer (PrCa) cell lines. The purpose of this study was to analyze morphogenetic properties of PrCa models in 3D, to compare phenotypes, gene expression and metabolism between 2D and 3D cultures, and to evaluate their relevance for pre-clinical drug discovery, disease modeling and basic research. Primary and non-transformed prostate epithelial cells, but also several PrCa lines, formed well-differentiated round spheroids. These showed strong cell-cell contacts, epithelial polarization, a hollow lumen and were covered by a complete basal lamina (BL). Most PrCa lines, however, formed large, poorly differentiated spheroids, or aggressively invading structures. In PC-3 and PC-3M cells, well-differentiated spheroids formed, which were then spontaneously transformed into highly invasive cells. These cell lines may have previously undergone an epithelial-to-mesenchymal transition (EMT), which is temporarily suppressed in favor of epithelial maturation by signals from the extracellular matrix (ECM). The induction of lipid and steroid metabolism, epigenetic reprogramming, and ECM remodeling represents a general adaptation to 3D culture, regardless of transformation and phenotype. In contrast, PI3-Kinase, AKT, STAT/interferon and integrin signaling pathways were particularly activated in invasive cells. Specific small molecule inhibitors targeted against PI3-Kinase blocked invasive cell growth more effectively in 3D than in 2D monolayer culture, or the growth of normal cells. Our panel of cell models, spanning a wide spectrum of phenotypic plasticity, supports the investigation of different modes of cell migration and tumor morphologies, and will be useful for predictive testing of anti-cancer and anti-metastatic compounds.
Härmä
Ville
V
Medical Biotechnology Knowledge Centre, VTT Technical Research Centre of Finland, Turku, Finland.
Virtanen
Johannes
J
Mäkelä
Rami
R
Happonen
Antti
A
Mpindi
John-Patrick
JP
Knuuttila
Matias
M
Kohonen
Pekka
P
Lötjönen
Jyrki
J
Kallioniemi
Olli
O
Nees
Matthias
M
D000970:Antineoplastic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002471:Cell Transformation, Neoplastic / Q000235:genetics / Q000473:pathology; D003094:Collagen / Q000494:pharmacology; D004338:Drug Combinations; D004847:Epithelial Cells / Q000187:drug effects / Q000473:pathology; D004848:Epithelium / Q000187:drug effects / Q000473:pathology; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors; D007797:Laminin / Q000494:pharmacology; D008297:Male; D008648:Mesoderm / Q000187:drug effects / Q000473:pathology; D008954:Models, Biological; D009361:Neoplasm Invasiveness; D009363:Neoplasm Proteins / Q000378:metabolism; D010641:Phenotype; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D025341:Principal Component Analysis; D011467:Prostate / Q000187:drug effects / Q000473:pathology; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000201:enzymology / Q000235:genetics / Q000473:pathology*; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors; D011509:Proteoglycans / Q000494:pharmacology; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D018874:Spheroids, Cellular / Q000187:drug effects / Q000201:enzymology / Q000473:pathology; D058570:TOR Serine-Threonine Kinases; D014407:Tumor Cells, Cultured
D000970:Antineoplastic Agents; D003094:Collagen; D004338:Drug Combinations; D047908:Intracellular Signaling Peptides and Proteins; D007797:Laminin; D009363:Neoplasm Proteins; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017346:Protein Serine-Threonine Kinases; D011509:Proteoglycans; D051057:Proto-Oncogene Proteins c-akt; D012333:RNA, Messenger; D058570:TOR Serine-Threonine Kinases; C059485:matrigel; C546842:MTOR protein, human
J Cell Biol. 2009 May 4;185(3):374-5
19414604
Breast Cancer Res Treat. 2010 Jul;122(1):35-43
19701706
J Biol Chem. 2009 Jan 9;284(2):1018-30
19001380
BMC Cancer. 2009 Nov 30;9:415
19948022
J Cell Biol. 1991 Dec;115(5):1383-95
1955479
Nat Methods. 2007 Apr;4(4):359-65
17396127
Cancer Res. 2006 Jul 15;66(14):7095-102
16849555
Genome Biol. 2008;9(5):R83
18492237
Stem Cells. 2009 Sep;27(9):2059-68
19544473
Development. 2001 Aug;128(16):3117-31
11688561
Mol Oncol. 2007 Jun;1(1):84-96
18516279
Oncogene. 2005 Nov 14;24(50):7465-74
16288293
Prostate. 2009 Nov 1;69(15):1683-93
19644960
Cell. 2008 May 16;133(4):704-15
18485877
PLoS Comput Biol. 2010 Feb 26;6(2):e1000684
20195492
Urology. 2006 Mar;67(3):647-53
16527597
Oncogene. 2009 Nov 5;28(44):3926-36
19684618
Prostate. 2003 Nov 1;57(3):205-25
14518029
J Mammary Gland Biol Neoplasia. 2010 Mar;15(1):65-71
20107877
Front Biosci. 2007 May 01;12:3468-74
17485314
Urol Oncol. 2010 Sep-Oct;28(5):473-9
19272800
Nat Clin Pract Oncol. 2008 May;5(5):280-90
18349857
J Clin Invest. 2009 Jun;119(6):1420-8
19487818
Prostate. 2009 Mar 1;69(4):428-42
19107869
Nat Rev Cancer. 2003 May;3(5):362-74
12724734
Cell. 2009 Nov 13;139(4):693-706
19878981
Cell. 2009 Aug 21;138(4):645-659
19682730
Cancer Res. 2008 May 15;68(10):3645-54
18483246
EMBO J. 2008 Nov 5;27(21):2829-38
18843297
Nat Cell Biol. 2007 Dec;9(12):1392-400
18037882
Int J Cancer. 2008 Dec 15;123(12):2774-81
18798265
Nat Cell Biol. 2009 Mar;11(3):257-68
19198601
Br J Cancer. 2009 Apr 7;100(7):1165-74
19277043
Cell Res. 2009 Jun;19(6):683-97
19365403
Cancer Res. 2006 May 1;66(9):4549-52
16651402
Nat Med. 2009 Sep;15(9):1010-2
19734877
Semin Cell Dev Biol. 2009 Oct;20(8):931-41
19682592
Cancer Res. 2007 Apr 1;67(7):2922-6
17409395
Cancer Res. 2008 Mar 1;68(5):1378-87
18316601
Breast Cancer Res. 2004;6(1):1-11
14680479
J Cell Biol. 1995 May;129(3):591-603
7730398
Cell Adh Migr. 2008 Oct-Dec;2(4):240-2
19262160
J Mammary Gland Biol Neoplasia. 2005 Jul;10(3):261-72
16807805
Methods Mol Biol. 2009;568:151-60
19582425
Dev Cell. 2008 Jun;14(6):818-29
18539112
J Cell Biol. 2003 Jan 20;160(2):267-77
12527751
PLoS One. 2008 Aug 20;3(8):e2994
18714348
Cancer Res. 2007 Sep 1;67(17):8131-8
17804725
Proc Natl Acad Sci U S A. 2009 Jan 6;106(1):268-73
19116269
Nature. 2008 Dec 18;456(7224):957-61
19078960
Cancer Res. 2006 Sep 1;66(17):8598-607
16951173
PLoS One. 2008;3(10):e3368
18852876
Cancer Res. 2007 Apr 1;67(7):3153-61
17409422
Clin Exp Metastasis. 2009;26(5):433-46
19221883
Proc Natl Acad Sci U S A. 2006 Mar 14;103(11):4134-9
16537497
J Cell Biol. 2010 Jan 11;188(1):11-9
19951899
Cancer Cell. 2002 Apr;1(3):279-88
12086864
Nat Genet. 2008 May;40(5):499-507
18443585
Cancer Res. 2008 Sep 15;68(18):7247-9
18794108
Cell Prolif. 2003 Oct;36 Suppl 1:59-72
14521516
Genome Biol. 2008;9(9):R139
18803840
J Natl Cancer Inst. 2006 Dec 20;98(24):1755-7
17179471
Nat Med. 2004 Jan;10(1):26-7
14702629
Gynecol Oncol. 2005 Apr;97(1):155-65
15790452
Cell. 2008 Oct 31;135(3):510-23
18984162