SciLifeLab, Department of Protein Science, KTH-Royal Institute of Technology, Solna, Sweden.
Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden.
Methods Mol Biol. 2022;2418:313-343. doi: 10.1007/978-1-0716-1920-9_18.
MicroRNAs play critical roles through their impact on posttranscriptional gene regulation. In cancer, they can act as oncogenes or tumor suppressors and can also function as biomarkers. Here, we describe a method for robust characterization of estrogen-regulated microRNA profiles. The activity of estrogen is mediated by two nuclear receptors, estrogen receptor alpha and estrogen receptor beta, and a transmembrane G-protein coupled estrogen receptor 1. This chapter details how to prepare cells for optimal estrogen response, directions for estrogen treatment, RNA extraction, different microRNA profiling approaches, and subsequent confirmations.
MicroRNAs 通过对转录后基因调控的影响发挥关键作用。在癌症中，它们可以作为癌基因或肿瘤抑制因子发挥作用，也可以作为生物标志物。在这里，我们描述了一种用于稳健表征雌激素调节的 microRNA 谱的方法。雌激素的活性由两种核受体，即雌激素受体 α 和雌激素受体 β，以及一种跨膜 G 蛋白偶联雌激素受体 1 介导。本章详细介绍了如何为最佳雌激素反应准备细胞，雌激素处理的方向，RNA 提取，不同的 microRNA 分析方法，以及随后的验证。
