Abstract
It is widely recognized that stromal fibroblasts significantly influence biological properties of multiple tumors including breast cancer. However, these epithelial–mesenchymal interactions seem to be essential in tumor biology and it is not fully clear whether this interaction is tumor type-specific or has a more general non-specific character. To elucidate this question, we tested the effect of cancer-associated fibroblasts (CAFs) isolated from different types of tumors (breast cancer skin metastasis, cutaneous basal cell carcinoma and melanoma, squamous cell carcinoma arising from oral cavity mucous membrane) on the EM-G3 breast cancer cell line. The results were compared with control experiments using normal human dermal fibroblasts, 3T3 mouse fibroblasts, and 3T3 fibroblasts influenced by the fibroblasts prepared from the basal cell carcinoma. Our results demonstrated that expression of luminal marker keratin 8 was influenced only by CAFs prepared from any tested tumors. In contrast, all tested types of fibroblasts showed a strong stimulatory effect on the expression of basal/myoepithelial marker keratin 14. The CAFs also elevated the number of cells with positivity for both keratins 8 and 14 that are similar to ductal originated precursor cells. The expression of proliferation marker Ki67 was not influenced by any of the tested fibroblasts. In conclusion, our data indicate that CAFs are able to influence the phenotype of a breast cancer cell line and this effect is based on a tumor type-unspecific mechanism. Finally, a clear functional difference between normal and CAFs was demonstrated.
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Acknowledgments
This study was supported by grant No. 304/12/1333 from the Grant Agency of the Czech Republic and by the Charles University project of University Center of Excellence and for support of Specific University Research No. 260510. Authors are grateful to Iva Burdová, Radana Kavková, and Vít Hajdúch for excellent technical assistance and to Dr. Šárka Takáčová for English revision.
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418_2012_918_MOESM1_ESM.tif
Suppl Fig. 1 Fibroblasts prepared from breast cancer skin metastasis as an example of employed fibroblasts. Cells with nuclear DNA marked by DAPI (A1) strongly exhibit vimentin (Vim, A2), and they are negative for keratins (K, A3). Bar represents 50 μm. (TIFF 167 kb)
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Suppl. Fig. 2 EM-G3 co-cultured with MELFF exhibit no K19 (red signal) after amplification of the signal. Bar represents 50 μm. (TIFF 384 kb)
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Suppl. Fig. 3 Detection of proliferation marker Ki67 (red signal) at EM-G3 cells positive for keratins (green signal) cultured without feeder cells (A) and in the presence of fibroblasts prepared from squamous cell carcinoma (SCCF, B). Quantitative analysis (C) demonstrated that there is no difference in the expression of Ki67 between EM-G3 cells cultured without feeder cells and those co-cultured with any type of the tested fibroblasts. (TIFF 5467 kb)
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Dvořánková, B., Szabo, P., Lacina, L. et al. Fibroblasts prepared from different types of malignant tumors stimulate expression of luminal marker keratin 8 in the EM-G3 breast cancer cell line. Histochem Cell Biol 137, 679–685 (2012). https://doi.org/10.1007/s00418-012-0918-3
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DOI: https://doi.org/10.1007/s00418-012-0918-3