Profiling of direct mRNA targets of miR-130a, miR-203 and miR-205 in prostate cancer cell line LNCaP. Homo sapiens

Micro RNAs (miRNAs) miR-130a, miR-203 and miR-205 are jointly downregulated in prostate cancer and act as repressors of AR-signaling. MiRNAs are small non-coding RNAs that regulate the expression of specific mRNA targets mainly by translational repression, mRNA deadenylation or cleavage. Reconstitution of these lost miRNAs in the LNCaP PCa cell line cause morphology changes, growth arrest, and apoptosis, increasing when the miRNAs were co-expressed. This series identifies direct targets of miR-130a, miR-203, and miR-205 by AGO2-RNA co-immunoprecipitation as described by (Beitzinger et al. 2007) upon miRNA reconstitution in LNCaP cells and analyzing AGO2-bound mRNAs using Affymetrix Genechips. Relative levels of AGO2 bound versus total RNA expression were compared between miRNA reconstituted and miR-scr transfected samples. Overall design: Three arrays each for AGO2-bound RNA upon reconstitution of miR-130a, miR-203, miR-205, a scramble miRNA, and three arrays each for total RNA upon reconstitution of miR-130a, miR-203, miR-205, a scramble miRNA.

微小核糖核酸（Micro RNAs，miRNAs）miR-130a、miR-203与miR-205在前列腺癌中共同下调，并作为雄激素受体（AR）信号通路的抑制因子发挥作用。miRNAs是一类小型非编码RNA，主要通过翻译抑制、mRNA脱腺苷化或mRNA裂解的方式调控特定靶mRNA的表达。在LNCaP前列腺癌（PCa）细胞系中恢复表达这些缺失的miRNAs，会引发细胞形态改变、生长停滞与细胞凋亡，且当这些miRNAs共表达时，上述效应会进一步增强。本数据集通过AGO2-RNA免疫共沉淀技术（参考Beitzinger等2007年的研究方法），在LNCaP细胞中恢复表达miRNAs后，利用Affymetrix Genechips基因芯片分析AGO2结合的mRNA，从而鉴定miR-130a、miR-203与miR-205的直接靶标。本研究对miRNA恢复表达组与miR-scr转染对照样本中，AGO2结合RNA的相对水平与总RNA表达量进行了比较。实验整体设计：针对miR-130a、miR-203、miR-205以及阴性对照scramble miRNA的恢复表达组，分别设置3个AGO2结合RNA样本的芯片检测重复；同时针对上述各组的总RNA样本，同样各设置3个芯片检测重复。