CRISPR screening of E3 ubiquitin ligases reveals Ring Finger Protein 185 as a novel tumor suppressor in glioblastoma repressed by promoter hypermethylation and miR-587

Journal of Translational Medicine

De Wei¹  Pengfeng Zheng²  Guangming Zeng²  Shizhong Wu²  Kun Lin²  Feng Lu²  Jingwei Liao²  Xiaohang Jiang²  Shang-Hang Shen³ 

[1] Department of Neurosurgery, Fujian Provincial Hospital South Branch, 516 Jinrong South Road, 350001, Fuzhou, China;Department of Neurosurgery, Fujian Provincial Hospital, 134 East Street, 350001, Fuzhou, China;Department of Neurosurgery, Shengli Clinical Medical College of Fujian Medical University, 134 East Street, 350001, Fuzhou, China;Department of Neurosurgery, Fujian Provincial Hospital South Branch, 516 Jinrong South Road, 350001, Fuzhou, China;Department of Neurosurgery, Shengli Clinical Medical College of Fujian Medical University, 516 Jinrong South Road, 350001, Fuzhou, China;Department of Neurosurgery, The First Affiliated Hospital of Xiamen University, Medical College of Xiamen University, 361003, Xiamen, China;

关键词: Glioblastoma;    CRISPR screening;    E3 ubiquitin ligases;    Ring Finger Protein 185;    Promoter hypermethylation;    miR-587;   

DOI  :  10.1186/s12967-022-03284-z

来源: Springer

【 摘 要 】

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.

【 授权许可】

CC BY