title: A bioengineered microenvironment to quantitatively measure the tumorigenic properties of cancer-associated fibroblasts in human prostate cancer.
abstract: Stromal-epithelial cell interactions play an important role in cancer and the tumor stroma is regarded as a therapeutic target. In vivo xenografting is commonly used to study cellular interactions not mimicked or quantified in conventional 2D culture systems. To interrogate the effects of tumor stroma (cancer-associated fibroblasts or CAFs) on epithelia, we created a bioengineered microenvironment using human prostatic tissues. Patient-matched CAFs and non-malignant prostatic fibroblasts (NPFs) from men with moderate (Gleason 7) and aggressive (Gleason 8-9 or castrate-resistant) prostate cancer were cultured with non-tumorigenic BPH-1 epithelial cells. Changes in the morphology, motility and phenotype of BPH-1 cells in response to CAFs and NPFs were analyzed using immunofluorescence and quantitative cell morphometric analyses. The matrix protein gene expression of CAFs, with proven tumorigenicity in vivo, had a significantly different gene expression profile of matrix proteins compared to patient matched NPFs. In co-culture with CAFs (but not NPFs), BPH-1 cells had a more invasive, elongated phenotype with increased motility and a more directed pattern of cell migration. CAFs from more aggressive tumors (Gleason 8-9 or CRPC) were not quantitatively different to moderate grade CAFs. Overall, our bioengineered microenvironment provides a novel 3D in vitro platform to systematically investigate the effects of tumor stroma on prostate cancer progression.
authors.affiliation: Department of Anatomy and Developmental Biology, School of Biomedical Sciences, Monash University, Clayton, Melbourne, Vic 3800, Australia.
authors.forename: Ashlee K
authors.initials: AK
authors.lastname: Clark
authors.forename: Anna V
authors.initials: AV
authors.lastname: Taubenberger
authors.forename: Renea A
authors.initials: RA
authors.lastname: Taylor
authors.forename: Birunthi
authors.initials: B
authors.lastname: Niranjan
authors.forename: Zhen Y
authors.initials: ZY
authors.lastname: Chea
authors.forename: Elena
authors.initials: E
authors.lastname: Zotenko
authors.forename: Shirly
authors.initials: S
authors.lastname: Sieh
authors.forename: John S
authors.initials: JS
authors.lastname: Pedersen
authors.forename: Sam
authors.initials: S
authors.lastname: Norden
authors.forename: Mark
authors.initials: M
authors.lastname: Frydenberg
authors.forename: Jeremy P
authors.initials: JP
authors.lastname: Grummet
authors.forename: David W
authors.initials: DW
authors.lastname: Pook
authors.forename: Clare
authors.initials: C
authors.lastname: Stirzaker
authors.forename: Susan J
authors.initials: SJ
authors.lastname: Clark
authors.forename: Mitchell G
authors.initials: MG
authors.lastname: Lawrence
authors.forename: Stuart J
authors.initials: SJ
authors.lastname: Ellem
authors.forename: Dietmar W
authors.initials: DW
authors.lastname: Hutmacher
authors.forename: Gail P
authors.initials: GP
authors.lastname: Risbridger
mesh_terms: D000328:Adult; D000368:Aged; D000818:Animals; D057005:Bioengineering / Q000379:methods*; D045744:Cell Line, Tumor; D002465:Cell Movement; D048430:Cell Shape; D002471:Cell Transformation, Neoplastic / Q000473:pathology*; D018920:Coculture Techniques; D018450:Disease Progression; D005347:Fibroblasts / Q000378:metabolism / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic; D052138:Genes, Neoplasm / Q000235:genetics; D006801:Humans; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D011470:Prostatic Hyperplasia / Q000378:metabolism / Q000473:pathology; D011471:Prostatic Neoplasms / Q000235:genetics / Q000473:pathology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017154:Stromal Cells / Q000473:pathology; D059008:Time-Lapse Imaging; D059016:Tumor Microenvironment