Reciprocal changes in gene expression profiles of cocultured breast epithelial cells and primary fibroblasts.
The importance of epithelial-stroma interaction in normal breast development and tumor progression has been recognized. To identify genes that were regulated by these reciprocal interactions, we cocultured a nonmalignant (MCF10A) and a breast cancer derived (MDA-MB231) basal cell lines, with fibroblasts isolated from breast benign-disease adjacent tissues (NAF) or with carcinoma-associated fibroblasts (CAF), in a transwell system. Gene expression profiles of each coculture pair were compared with the correspondent monocultures, using a customized microarray. Contrariwise to large alterations in epithelial cells genomic profiles, fibroblasts were less affected. In MDA-MB231 highly represented genes downregulated by CAF derived factors coded for proteins important for the specificity of vectorial transport between ER and golgi, possibly affecting cell polarity whereas the response of MCF10A comprised an induction of genes coding for stress responsive proteins, representing a prosurvival effect. While NAF downregulated genes encoding proteins associated to glycolipid and fatty acid biosynthesis in MDA-MB231, potentially affecting membrane biogenesis, in MCF10A, genes critical for growth control and adhesion were altered. NAFs responded to coculture with MDA-MB231 by a decrease in the expression of genes induced by TGFbeta1 and associated to motility. However, there was little change in NAFs gene expression profile influenced by MCF10A. CAFs responded to the presence of both epithelial cells inducing genes implicated in cell proliferation. Our data indicate that interactions between breast fibroblasts and basal epithelial cells resulted in alterations in the genomic profiles of both cell types which may help to clarify some aspects of this heterotypic signaling.
Rozenchan
Patricia Bortman
PB
Disciplina de Oncologia, Departamento de Radiologia, Faculdade de Medicina da Universidade de São Paulo, Hospital A.C. Camargo, Av. Dr. Arnaldo, 455, Sala 4112, São Paulo, SP CEP 01246-903, Brazil.
Carraro
Dirce Maria
DM
Brentani
Helena
H
de Carvalho Mota
Louise Danielle
LD
Bastos
Elen Pereira
EP
e Ferreira
Elisa Napolitano
EN
Torres
Cesar H
CH
Katayama
Maria Lúcia Hirata
ML
Roela
Rosimeire Aparecida
RA
Lyra
Eduardo C
EC
Soares
Fernando Augusto
FA
Folgueira
Maria Aparecida Azevedo Koike
MA
Góes
João Carlos Guedes Sampaio
JC
Brentani
Maria Mitzi
MM
D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism; D015153:Blotting, Western; D001940:Breast / Q000166:cytology; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology*; D049109:Cell Proliferation; D018920:Coculture Techniques; D004847:Epithelial Cells / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism*; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D009363:Neoplasm Proteins / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D012333:RNA, Messenger