Differential microRNA expression in adherent versus suspended MCF10A mammary epithelial cells

The ability to survive following loss of adhesion underpins cancer metastasis and significantly contributes triple negative breast cancer (TNBC) morbidity. Epithelial cells that form solid tumors must evade anoikis, a special type of cell death that occurs upon detachment from the extracellular matrix in order to colonize in distal sites. While there have been several advances in the understanding of the mechanisms by which cells undergo anoikis, the role for noncoding RNAs (ncRNAs) in this process is less well understood. An important role for microRNAs (miRNAs) in regulating anoikis is beginning to emerge, however in general the mechanisms by which miRNAs regulate anoikis are largely unknown. Thus we utilized the anoikis-sensitive immortalized mammary epithelial cell line, MCF10A to unbiasedly determine the microRNAs differentially regulated upon detachment. Overall design: Total RNA (50 µg) was extracted from MCF10A cultured for 24 hours on uncoated tissue culture plates (attached) or on tissue culture plates coated with 2% Poly(2-hydroxyethyl methacrylate) in 95% ethanol(detached). Small RNAs 18-24 nucleotides in length were size selected and gel purified.