Abstract
Molecular signaling through both estrogen and microRNAs are critical for breast cancer development and growth. The activity of estrogen is mediated by transcription factors, the estrogen receptors. Here we describe a method for robust characterization of estrogen-regulated microRNA profiles. The method details how to prepare cells for optimal estrogen response, directions for estrogen treatment, RNA extraction, microRNA large-scale profiling, and subsequent confirmations.
Keywords:
Breast cancer; Estrogen; Estrogen receptor; MicroRNA; Microarray; qPCR.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Breast Neoplasms / genetics*
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Breast Neoplasms / metabolism
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Estradiol / pharmacology*
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Estrogen Receptor alpha / agonists
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Estrogen Receptor alpha / metabolism
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Estrogens / pharmacology*
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Female
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Gene Expression Profiling / methods*
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Gene Expression Regulation, Neoplastic / drug effects*
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Humans
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MCF-7 Cells
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MicroRNAs / genetics*
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MicroRNAs / metabolism
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Oligonucleotide Array Sequence Analysis*
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Real-Time Polymerase Chain Reaction
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Reproducibility of Results
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Time Factors
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Workflow
Substances
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ESR1 protein, human
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Estrogen Receptor alpha
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Estrogens
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MicroRNAs
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Estradiol