Profiling and functional analysis of microRNAs in follicular granulosa cells and their clinical application

我们在人粒层细胞来源的细胞系（例如，KGN）。在这些细胞系中，我们发现miR-21占AGO 2（命名为EIFC 2）结合的miRNA的80%以上，表明它在人类GC中作为功能性miRNA发挥着核心作用。我们还在KGN细胞中进行了AGO 2结合mRNA的cDNA克隆，并将COL 4A 1 mRNA鉴定为KGN细胞中可能的miR-21靶标，随后的实验验证证实了这一点。在使用miRNA前体或短干扰RNA的合成模拟物进行的体外功能研究中，我们还发现这些短双链RNA（23-和/或24-核苷酸dsRNA）容易诱导人颗粒细胞肿瘤衍生细胞的凋亡，但不诱导其他细胞类型的凋亡。此外，我们发现，短的双链RNA转录超诱导维甲酸诱导基因-I通过双链RNA激活的蛋白激酶。

We performed large-scale profiling of microRNA (miRNAs) in human granulosa-derived cell lines (e.g., KGN) by high-throughput sequencing. In these cell lines, we found that miR-21 accounted for more than 80% of AGO2 (designated EIFC2) -bound miRNAs, suggesting that it played a central role as a functional miRNA in human GCs. We also performed cDNA cloning of AGO2-bound mRNAs in KGN cells and identified COL4A1 mRNA as a possible miR-21 target in KGN cells, which was corroborated by subsequent experimental validation. In proceeding in vitro functional studies using synthetic mimics of miRNA precursors or short-interference RNAs, we also found that these short double-strand RNAs (23- and/or 24-nucleotide dsRNAs) readily induced apoptosis in human granulosa cell tumor-derived cells but not in other cell types. Furthermore, we revealed that the short dsRNAs transcriptionally hyper-induce retinoic acid-inducible gene-I through dsRNA-activated protein kinase.