Department of Neurosurgery, Fujian Provincial Hospital South Branch, 516 Jinrong South Road, Fuzhou, 350001, China.
Department of Neurosurgery, Shengli Clinical Medical College of Fujian Medical University, 516 Jinrong South Road, Fuzhou, 350001, China.
J Transl Med. 2022 Feb 19;20(1):96. doi: 10.1186/s12967-022-03284-z.
Glioblastoma (GBM) is the most common and aggressive malignant primary brain tumor. E3 ligases play important functions in glioma pathogenesis. CRISPR system offers a powerful platform for genome manipulation, while the screen of E3 ligases in GBM still remains to be explored. Here, we first constructed an E3 ligase small guide RNA (sgRNAs) library for glioma cells growth screening. After four passages, 299 significantly enriched or lost genes (SELGs) were compared with the initial state. Then the clinical significance of SELGs were validated and analyzed with TCGA glioblastoma and CGGA datasets. As RNF185 showed lost signal, decreased expression and favorable prognostic significance, we chose RNF185 for functional analysis. In vitro overexpressed cellular phenotype showed that RNF185 was a tumor suppressor in two glioma cell lines. Finally, the molecular mechanism of decreased RNF185 expression was investigated and increased miR-587 expression and DNA hypermethylation was evaluated. This study would provide a link between the molecular basis and glioblastoma pathogenesis, and a novel perspective for glioblastoma treatment.
胶质母细胞瘤（GBM）是最常见和侵袭性最强的原发性脑肿瘤。E3 连接酶在神经胶质瘤发病机制中发挥着重要作用。CRISPR 系统为基因组操作提供了一个强大的平台，而 GBM 中 E3 连接酶的筛选仍有待探索。在这里，我们首先构建了一个用于神经胶质瘤细胞生长筛选的 E3 连接酶小向导 RNA（sgRNA）文库。经过四个传代，与初始状态相比，有 299 个明显富集或丢失的基因（SELGs）被筛选出来。然后，我们利用 TCGA 胶质母细胞瘤和 CGGA 数据集验证和分析了 SELGs 的临床意义。由于 RNF185 表现出丢失信号、表达降低和有利的预后意义，我们选择 RNF185 进行功能分析。体外过表达细胞表型表明，RNF185 在两种神经胶质瘤细胞系中是一种肿瘤抑制因子。最后，研究了 RNF185 表达降低的分子机制，并评估了 miR-587 表达增加和 DNA 超甲基化。本研究将为神经胶质瘤发病机制的分子基础提供联系，并为神经胶质瘤的治疗提供新的视角。
