MDA-MB-231 VIM RFP
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Mesenchymal to epithelial transition, anti-EMT drug screening, metastatic breast cancer drug screening, vimentin intermediate filament dynamics. See Technical Data Sheet.
The MDA-MB-231 VIM RFP reporter cell line provides a convenient and sensitive platform for research on the mechanisms of metastasis in vitro and the development of new anti-EMT drugs for metastatic breast cancer.
Breast cancer is the most aggressive form of all cancers, with high incidence and mortality rates. Although epithelial-to-mesenchymal transition (EMT) and mesenchymal-to-epithelial transition (MET) have been implicated in the incidence of cancer metastasis and drug resistance, their impact in cancer progression and patient survival is not fully understood (NIETO et al. 2016). During EMT, epithelial cells lose their polarity, as well as their cell-cell adhesions, and acquire the motile and invasive characteristics of mesenchymal cells (HAY 1995). Proteins such as vimentin (VIM) intermediate filament (IF) are generally upregulated when the cell is in the mesenchymal relative to the epithelial status (GILLES et al. 1999; THIERY and SLEEMAN 2006; RICHARDSON et al. 2012; LAMOUILLE et al. 2014).
The VIM RFP reporter cell line (ATCC HTB-26MET) was created using CRISPR/Cas9 gene editing and the parental MDA-MB-231 breast adenocarcinoma cell line (ATCC HTB-26). HTB-26MET harbors a C-terminal red fluorescent protein (RFP) tag on the vimentin gene. This enables the tracking of the EMT status of cells in vitro by monitoring RFP expression. The integrity of the VIM RFP knock-in has been verified at the genomic, mRNA, and protein level for sequence and expression. Functional evaluation of HTB-26MET shows sensitivity to metastatic breast cancer drugs axitinib (tyrosine kinase inhibitor) and U0126 (MEK1/2 inhibitor) via the inhibition of the inherent signaling pathways which impact EMT.
The base medium for this cell line is Eagle's Minimum Essential Medium (EMEM; ATCC 30-2003). To make the complete medium, add the following components to the base medium at the indicated final concentrations:
To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability.
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Thiery JP, Sleeman JP. Complex networks orchestrate epithelial-mesenchymal transitions. Nat Rev Mol Cell Biol 7(2): 131-142, 2006. PubMed: 16493418
Richardson F, et al. The evaluation of E-Cadherin and vimentin as biomarkers of clinical outcomes among patients with non-small cell lung cancer treated with erlotinib as second- or third-line therapy. Anticancer Res 32(2): 537-552, 2012. PubMed: 22287743
Gilles C, et al. Vimentin contributes to human mammary epithelial cell migration. J Cell Sci 112 (Pt 24): 4615-4625, 1999. PubMed: 10574710