title: Characteristic Gene Expression Profiles of Human Fibroblasts and Breast Cancer Cells in a Newly Developed Bilateral Coculture System.
abstract: The microenvironment of cancer cells has been implicated in cancer development and progression. Cancer-associated fibroblast constitutes a major stromal component of the microenvironment. To analyze interaction between cancer cells and fibroblasts, we have developed a new bilateral coculture system using a two-sided microporous collagen membrane. Human normal skin fibroblasts were cocultured with three different human breast cancer cell lines: MCF-7, SK-BR-3, and HCC1937. After coculture, mRNA was extracted separately from cancer cells and fibroblasts and applied to transcriptomic analysis with microarray. Top 500 commonly up- or downregulated genes were characterized by enrichment functional analysis using MetaCore Functional Analysis. Most of the genes upregulated in cancer cells were downregulated in fibroblasts while most of the genes downregulated in cancer cells were upregulated in fibroblasts, indicating that changing patterns of mRNA expression were reciprocal between cancer cells and fibroblasts. In coculture, breast cancer cells commonly increased genes related to mitotic response and TCA pathway while fibroblasts increased genes related to carbohydrate metabolism including glycolysis, glycogenesis, and glucose transport, indicating that fibroblasts support cancer cell proliferation by supplying energy sources. We propose that the bilateral coculture system using collagen membrane is useful to study interactions between cancer cells and stromal cells by mimicking in vivo tumor microenvironment.
authors.affiliation: Department of Breast Surgery, Kyoto University Hospital, Kyoto 606 8507, Japan ; Department of Breast Surgery, Kyorin University Hospital, Tokyo 181 8611, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Ueno
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606 8501, Japan ; Cancer Institute, Japanese Foundation for Cancer Research, Tokyo 135 8550, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Utsumi
authors.affiliation: Department of Breast Surgery, Kyoto University Hospital, Kyoto 606 8507, Japan.
authors.forename: Masakazu
authors.initials: M
authors.lastname: Toi
authors.affiliation: Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606 8501, Japan.
authors.forename: Kazuharu
authors.initials: K
authors.lastname: Shimizu
mesh_terms: D001943:Breast Neoplasms / Q000378:metabolism*; D045744:Cell Line, Tumor; D018920:Coculture Techniques / Q000379:methods*; D015536:Down-Regulation; D005260:Female; D005347:Fibroblasts / Q000378:metabolism*; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D011506:Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D059467:Transcriptome / Q000235:genetics; D015854:Up-Regulation
chemical_list: D011506:Proteins