Induction of Epithelial to Mesenchymal Transition in PMC42-LA Human Breast Carcinoma Cells by Carcinoma-associated Fibroblast Secreted Factors

Overview

Journal Breast Cancer Res

Specialty Oncology

Date 2007 Feb 22

PMID 17311675

Citations 51

Authors

Stephanie C Lebret

Donald F Newgreen

Erik W Thompson

M Leigh Ackland

Affiliations

Soon will be listed here.

Abstract

Background: Breast carcinoma is accompanied by changes in the acellular and cellular components of the microenvironment, the latter typified by a switch from fibroblasts to myofibroblasts.

Methods: We utilised conditioned media cultures, Western blot analysis and immunocytochemistry to investigate the differential effects of normal mammary fibroblasts (NMFs) and mammary cancer-associated fibroblasts (CAFs) on the phenotype and behaviour of PMC42-LA breast cancer cells. NMFs were obtained from a mammary gland at reduction mammoplasty, and CAFs from a mammary carcinoma after resection.

Results: We found greater expression of myofibroblastic markers in CAFs than in NMFs. Medium from both CAFs and NMFs induced novel expression of alpha-smooth muscle actin and cytokeratin-14 in PMC42-LA organoids. However, although conditioned media from NMFs resulted in distribution of vimentin-positive cells to the periphery of PMC42-LA organoids, this was not seen with CAF-conditioned medium. Upregulation of vimentin was accompanied by a mis-localization of E-cadherin, suggesting a loss of adhesive function. This was confirmed by visualizing the change in active beta-catenin, localized to the cell junctions in control cells/cells in NMF-conditioned medium, to inactive beta-catenin, localized to nuclei and cytoplasm in cells in CAF-conditioned medium.

Conclusion: We found no significant difference between the influences of NMFs and CAFs on PMC42-LA cell proliferation, viability, or apoptosis; significantly, we demonstrated a role for CAFs, but not for NMFs, in increasing the migratory ability of PMC42-LA cells. By concentrating NMF-conditioned media, we demonstrated the presence of factor(s) that induce epithelial-mesenchymal transition in NMF-conditioned media that are present at higher levels in CAF-conditioned media. Our in vitro results are consistent with observations in vivo showing that alterations in stroma influence the phenotype and behaviour of surrounding cells and provide evidence for a role for CAFs in stimulating cancer progression via an epithelial-mesenchymal transition. These findings have implications for our understanding of the roles of signalling between epithelial and stromal cells in the development and progression of mammary carcinoma.

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References

Luo J, Lubaroff D, Hendrix M . Suppression of prostate cancer invasive potential and matrix metalloproteinase activity by E-cadherin transfection. Cancer Res. 1999; 59(15):3552-6. View

Olumi A, Grossfeld G, Hayward S, Carroll P, Tlsty T, Cunha G . Carcinoma-associated fibroblasts direct tumor progression of initiated human prostatic epithelium. Cancer Res. 1999; 59(19):5002-11. PMC: 3300837. DOI: 10.1186/bcr138. View

Bhowmick N, Neilson E, Moses H . Stromal fibroblasts in cancer initiation and progression. Nature. 2004; 432(7015):332-7. PMC: 3050735. DOI: 10.1038/nature03096. View

Sadlonova A, Novak Z, Johnson M, Bowe D, Gault S, Page G . Breast fibroblasts modulate epithelial cell proliferation in three-dimensional in vitro co-culture. Breast Cancer Res. 2005; 7(1):R46-59. PMC: 1064098. DOI: 10.1186/bcr949. View

Barcellos-Hoff M, Medina D . New highlights on stroma-epithelial interactions in breast cancer. Breast Cancer Res. 2005; 7(1):33-6. PMC: 1064117. DOI: 10.1186/bcr972. View

Van Roozendaal K, Klijn J, van Ooijen B, Claassen C, Eggermont A, Henzen-Logmans S . Differential regulation of breast tumor cell proliferation by stromal fibroblasts of various breast tissue sources. Int J Cancer. 1996; 65(1):120-5. DOI: 10.1002/(SICI)1097-0215(19960103)65:1<120::AID-IJC20>3.0.CO;2-J. View

Chen H, Paradies N, Brackenbury R . E-cadherin mediates adhesion and suppresses cell motility via distinct mechanisms. J Cell Sci. 1997; 110 ( Pt 3):345-56. DOI: 10.1242/jcs.110.3.345. View

Yashiro M, Ikeda K, Tendo M, Ishikawa T, Hirakawa K . Effect of organ-specific fibroblasts on proliferation and differentiation of breast cancer cells. Breast Cancer Res Treat. 2005; 90(3):307-13. DOI: 10.1007/s10549-004-5364-z. View

Xian W, Schwertfeger K, Vargo-Gogola T, Rosen J . Pleiotropic effects of FGFR1 on cell proliferation, survival, and migration in a 3D mammary epithelial cell model. J Cell Biol. 2005; 171(4):663-73. PMC: 2171554. DOI: 10.1083/jcb.200505098. View

10.

Bellovin D, Bates R, Muzikansky A, Rimm D, Mercurio A . Altered localization of p120 catenin during epithelial to mesenchymal transition of colon carcinoma is prognostic for aggressive disease. Cancer Res. 2005; 65(23):10938-45. DOI: 10.1158/0008-5472.CAN-05-1947. View

11.

Kuroda H, Tamaru J, Takeuchi I, Ohnisi K, Sakamoto G, Adachi A . Expression of E-cadherin, alpha-catenin, and beta-catenin in tubulolobular carcinoma of the breast. Virchows Arch. 2006; 448(4):500-5. DOI: 10.1007/s00428-005-0096-y. View

12.

Lebret S, Newgreen D, Waltham M, Price J, Thompson E, Ackland M . Myoepithelial molecular markers in human breast carcinoma PMC42-LA cells are induced by extracellular matrix and stromal cells. In Vitro Cell Dev Biol Anim. 2007; 42(10):298-307. DOI: 10.1290/0601004.1. View

13.

Darcy K, Zangani D, Shoemaker S, Lee P, Mead L, Mudipalli A . Mammary fibroblasts stimulate growth, alveolar morphogenesis, and functional differentiation of normal rat mammary epithelial cells. In Vitro Cell Dev Biol Anim. 2001; 36(9):578-92. DOI: 10.1007/BF02577526. View

14.

Ackland M, Michalczyk A, Whitehead R . PMC42, a novel model for the differentiated human breast. Exp Cell Res. 2001; 263(1):14-22. DOI: 10.1006/excr.2000.5106. View

15.

Petersen O, Lind Nielsen H, Gudjonsson T, Villadsen R, Ronnov-Jessen L, Bissell M . The plasticity of human breast carcinoma cells is more than epithelial to mesenchymal conversion. Breast Cancer Res. 2001; 3(4):213-7. PMC: 138684. DOI: 10.1186/bcr298. View

16.

Nielsen B, Rank F, Lopez J, Balbin M, Vizoso F, Lund L . Collagenase-3 expression in breast myofibroblasts as a molecular marker of transition of ductal carcinoma in situ lesions to invasive ductal carcinomas. Cancer Res. 2001; 61(19):7091-100. View

17.

Berx G, VAN Roy F . The E-cadherin/catenin complex: an important gatekeeper in breast cancer tumorigenesis and malignant progression. Breast Cancer Res. 2001; 3(5):289-93. PMC: 138690. DOI: 10.1186/bcr309. View

18.

Hayward S, Wang Y, Cao M, Hom Y, Zhang B, Grossfeld G . Malignant transformation in a nontumorigenic human prostatic epithelial cell line. Cancer Res. 2001; 61(22):8135-42. View

19.

Bissell M, Radisky D . Putting tumours in context. Nat Rev Cancer. 2002; 1(1):46-54. PMC: 2975572. DOI: 10.1038/35094059. View

20.

Thiery J . Epithelial-mesenchymal transitions in tumour progression. Nat Rev Cancer. 2002; 2(6):442-54. DOI: 10.1038/nrc822. View