An Effect of Culture Media on Epithelial Differentiation Markers in Breast Cancer Cell Lines MCF7, MDA-MB-436 and SkBr3

Medicina (Lithuania) - Tập 54 Số 2 - Trang 11
Valdis Pirsko1, Inese Čakstiņa1, Marta Priedite1, Rasma Dortane1, Linda Feldmane1, Miki Nakazawa1, Zanda Daneberga1, Jānis Gardovskis1, Edvīns Miklaševičs1
1Institute of Oncology, Riga Stradins University, LV1086 Riga, Latvia

Tóm tắt

Background and objectives: Cell culture is one of the mainstays in the research of breast cancer biology, although the extent to which this approach allows to preserve the original characteristics of originating tumor and implications of cell culture findings to real life situations have been widely debated in the literature. The aim of this study was to determine the role of three cell culture media on transcriptional expression of breast cancer markers in three breast cancer reference cell lines (MCF7, SkBr3 and MDA-MB-436). Materials and methods: Cell lines were conditioned in three studied media (all containing 5% fetal bovine serum (FBS) + hormones/growth factors; different composition of basal media) for four passages. Population growth was characterized by cumulative population doubling levels, average generation time, cell yield and viability at the fourth passage. Transcriptional expression of breast cancer differentiation markers and regulatory transcriptional programs was measured by qPCR. Results: Differences in the composition of growth media significantly influenced the growth of studied cell lines and the expression of mammary lineage governing transcriptional programs and luminal/basal markers. Effects of media on transcriptional expression were more pronounced in luminal cell lines (MCF7, SkBr3), than in the basal cell line (MDA-MB-436). Changes in growth media in terms of supplementation and basal medium delayed growth of cells, but improved cell yields. Conclusions: The expression of breast cancer cell differentiation phenotypic markers depends on the composition of cell growth medium, therefore cell culture as a tool in phenotypic studies should be used considering this effect. The findings of such studies should always be interpreted with caution. The formulation of cell growth media has greater effect on the expression of phenotypic markers in luminal, rather than basal cell lines. Media containing mitogens and higher vitamin content improved efficacy of cell culture in terms of cell yields, although greatly increased growth times.

Từ khóa


Tài liệu tham khảo

Rosen, 2007, Modelling breast cancer: One size does not fit all, Nat. Rev. Cancer, 7, 659, 10.1038/nrc2193

Holliday, 2011, Choosing the right cell line for breast cancer research, Breast Cancer Res., 13, 215, 10.1186/bcr2889

Eccles, 2013, Critical research gaps and translational priorities for the successful prevention and treatment of breast cancer, Breast Cancer Res., 15, R92, 10.1186/bcr3493

Lacroix, 2004, Relevance of breast cancer cell lines as models for breast tumors: An update, Breast Cancer Res. Treat., 83, 249, 10.1023/B:BREA.0000014042.54925.cc

Vinci, M., Gowan, S., Boxall, F., Patterson, L., Zimmermann, M., Court, W., Lomas, C., Mendiola, M., Hardisson, D., and Eccles, S.A. (2012). Advances in establishment and analysis of three-dimensional tumor spheroid-based functional assays for target validation and drug evaluation. BMC Biol., 10.

Prat, 2013, Characterization of cell lines derived from breast cancers and normal mammary tissues for the study of the intrinsic molecular subtypes, Breast Cancer Res. Treat., 142, 237, 10.1007/s10549-013-2743-3

Neve, 2006, A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes, Cancer Cell, 10, 515, 10.1016/j.ccr.2006.10.008

Lim, 2009, Aberrant luminal progenitors as the candidate target population for basal tumor development in BRCA1 mutation carriers, Nat. Med., 15, 907, 10.1038/nm.2000

Shoemaker, 2006, The NCI60 human tumor cell line anticancer drug screen, Nat. Rev. Cancer, 6, 813, 10.1038/nrc1951

Monks, 1991, Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines, J. Natl. Cancer Inst., 83, 757, 10.1093/jnci/83.11.757

Prat, 2011, Deconstructing the molecular portraits of breast cancer, Mol. Oncol., 5, 5, 10.1016/j.molonc.2010.11.003

Sorlie, 2001, Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications, Proc. Natl. Acad. Sci. USA, 98, 10869, 10.1073/pnas.191367098

Hu, Z., Fan, C., Oh, D.S., Marron, J.S., He, X., Qaqish, B.F., Livasy, C., Carey, L.A., Reynolds, E., and Dressler, L. (2006). The molecular portraits of breast tumors are conserved across microarray platforms. BMC Genom., 7.

Prat, 2010, Phenotypic and molecular characterization of the claudin-low intrinsic subtype of breast cancer, Breast Cancer Res., 12, R68, 10.1186/bcr2635

Mackay, 2009, A high-resolution integrated analysis of genetic and expression profiles of breast cancer cell lines, Breast Cancer Res. Treat., 118, 481, 10.1007/s10549-008-0296-7

Kao, J., Salari, K., Bocanegra, M., Choi, Y.-L., Girard, L., Gandhi, J., Kwei, K.A., Hernandez-Boussard, T., Wang, P., and Gazdar, A.F. (2009). Molecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PLoS ONE, 4.

Ross, 2001, A comparison of gene expression signatures from breast tumors and breast tissue derived cell lines, Dis. Mark., 17, 99, 10.1155/2001/850531

Perou, 2000, Molecular portraits of human breast tumors, Nature, 406, 747, 10.1038/35021093

Parker, 2009, Supervised risk predictor of breast cancer based on intrinsic subtypes, J. Clin. Oncol., 27, 1160, 10.1200/JCO.2008.18.1370

Rocha, 2011, Distinct stem cells contribute to mammary gland development and maintenance, Nature, 479, 189, 10.1038/nature10573

Shehata, 2012, Phenotypic and functional characterisation of the luminal cell hierarchy of the mammary gland, Breast Cancer Res., 14, R134, 10.1186/bcr3334

Granit, R.Z., Slyper, M., and Ben-Porath, I. (2014). Axes of Differentiation in Breast Cancer: Untangling Stemness, Lineage Identity, and the Epithelial to Mesenchymal Transition, John Wiley & Sons, Inc.. Wiley Interdisciplinary Reviews. Systems Biology and Medicine.

Chou, 2010, GATA3 in development and cancer differentiation: Cells GATA have it!, J. Cell. Physiol., 222, 42, 10.1002/jcp.21943

Kim, 2008, GATA-3 and the regulation of the mammary luminal cell fate, Curr. Opin. Cell Biol., 20, 164, 10.1016/j.ceb.2008.02.003

Douglas, 2013, The T-box transcription factors TBX2 and TBX3 in mammary gland development and breast cancer, J. Mammary Gland Biol. Neoplasia, 18, 143, 10.1007/s10911-013-9282-8

Bouras, 2008, Notch signaling regulates mammary stem cell function and luminal cell-fate commitment, Cell Stem Cell, 3, 429, 10.1016/j.stem.2008.08.001

Visvader, 2014, Mammary stem cells and the differentiation hierarchy: Current status and perspectives, Genes Dev., 28, 1143, 10.1101/gad.242511.114

Heiser, 2012, Subtype and pathway specific responses to anticancer compounds in breast cancer, Proc. Natl. Acad. Sci. USA, 109, 2724, 10.1073/pnas.1018854108

Niepel, 2013, Profiles of basal and stimulated receptor signaling networks predict drug response in breast cancer lines, Sci Signal., 6, ra84, 10.1126/scisignal.2004379

Liu, 2012, ROCK inhibitor and feeder cells induce the conditional reprogramming of epithelial cells, Am. J. Pathol., 180, 599, 10.1016/j.ajpath.2011.10.036

Ethier, 1993, Differential isolation of normal luminal mammary epithelial cells and breast cancer cells from primary and metastatic sites using selective media, Cancer Res., 53, 627

Ip, M.M., and Asch, B.B. (2000). Isolation and Culture of Human Breast Cancer Cells from Primary Tumors and Metastases. Methods in Mammary Gland Biology and Breast Cancer Research, Springer.

Wallden, B., Storhoff, J., Nielsen, T., Dowidar, N., Schaper, C., Ferree, S., Liu, S., Leung, S., Geiss, G., and Snider, J. (2015). Development and verification of the PAM50-based Prosigna breast cancer gene signature assay. BMC Med. Genom., 8.

Waltz, 2007, Estrogen promotes reversible epithelial-to-mesenchymal-like transition and collective motility in MCF-7 breast cancer cells, J. Steroid Biochem. Mole. Biol., 104, 11, 10.1016/j.jsbmb.2006.09.039

Du, 2010, Overexpression of DeltaNp63alpha induces a stem cell phenotype in MCF7 breast carcinoma cell line through the Notch pathway, Cancer Sci., 101, 2417, 10.1111/j.1349-7006.2010.01700.x

Subramanyam, 2011, Notch signaling pathway as a therapeutic target in breast cancer, Mol. Cancer Ther., 10, 9, 10.1158/1535-7163.MCT-10-0677

Ouzounova, 2015, Notch reporter activity in breast cancer cell lines identifies a subset of cells with stem cell activity. Molecular Cancer, Therapeutics, 14, 779

Mohammed, 2015, Progesterone receptor modulates ERalpha action in breast cancer, Nature, 523, 313, 10.1038/nature14583

Zheng, 2005, A novel antiestrogenic mechanism in progesterone receptor-transfected breast cancer cells, J. Biol. Chem., 280, 17480, 10.1074/jbc.M501261200

Fan, 2004, TBX3 and its isoform TBX3+2a are functionally distinctive in inhibition of senescence and are overexpressed in a subset of breast cancer cell lines, Cancer Res., 64, 5132, 10.1158/0008-5472.CAN-04-0615

Fillmore, 2010, Estrogen expands breast cancer stem-like cells through paracrine FGF/Tbx3 signaling, Proc. Natl. Acad. Sci. USA, 107, 21737, 10.1073/pnas.1007863107

Pendse, 2017, Information-dependent enrichment analysis reveals time-dependent transcriptional regulation of the estrogen pathway of toxicity, Arch. Toxicol., 91, 1749, 10.1007/s00204-016-1824-6

Stender, 2007, Estrogen-Regulated Gene Networks in Human Breast Cancer Cells: Involvement of E2F1 in the Regulation of Cell Proliferation, Mol. Endocrinol., 21, 2112, 10.1210/me.2006-0474

Frasor, 2003, Profiling of Estrogen Up- and Down-Regulated Gene Expression in Human Breast Cancer Cells: Insights into Gene Networks and Pathways Underlying Estrogenic Control of Proliferation and Cell Phenotype, Endocrinology, 144, 4562, 10.1210/en.2003-0567

Nagai, 2015, Gene Expression Profiles in Breast Cancer to Identify Estrogen Receptor Target Genes, Recent Adv. Med. Chem., 1, 381

Yamaga, 2013, RNA sequencing of MCF-7 breast cancer cells identifies novel estrogen-responsive genes with functional estrogen receptor-binding sites in the vicinity of their transcription start sites, Hormones Cancer, 4, 222, 10.1007/s12672-013-0140-3

Omoto, 2002, A study of estrogen signaling using DNA microarray in human breast cancer, Breast Cancer, 9, 308, 10.1007/BF02967609

Thông tin
Thông tin xuất bản

Medicina (Lithuania)

Tập 54 Số 2

11

Thông tin tác giả