UBE2T 是一种预后生物标志物，与视网膜母细胞瘤中的 Th2 细胞浸润相关。

UBE2T is a prognostic biomarker and correlated with Th2 cell infiltrates in retinoblastoma.

机构信息

The Fourth School of Clinical Medicine, The Affiliated Eye Hospital, Nanjing Medical University, Nanjing, China.

Shanghai Medical College, Eye and ENT Hospital, Fudan University, Shanghai, China.

出版信息

Biochem Biophys Res Commun. 2022 Jul 23;614:138-144. doi: 10.1016/j.bbrc.2022.04.096. Epub 2022 May 6.

DOI:10.1016/j.bbrc.2022.04.096

PMID:35594577

Abstract

OBJECTIVE

This study aimed to screen anaplasia-related genes that influence the progression of retinoblastoma (RB) and to identify immune cells associated with the poor prognosis.

METHODS

Differentially expressed genes (DEGs) between retina and RB samples were acquired from gene expression omnibus (GEO) database. Candidate hub genes were screened by taking intersections among the co-expressed genes, the hub nodes, and DEGs of the validation set. The hub genes were identified by receiver operating characteristic (ROC) and quantitative real-time PCR (qPCR). Immune infiltration levels of RB tissues were estimated using single-sample gene set enrichment analysis (ssGSEA). The functions of RB cells were detected by CCK8, EDU and flow cytometry assays.

RESULTS

665 DEGs involved in the genesis and progression of RB were acquired from GEO database. 29 candidate hub genes were screened by examining 43 co-expressed genes and 63 hub nodes. 9 hub genes (CHEK1, EXO1, FANCI, GTSE1, MELK, MKI67, NCAPH, PRC1, and UBE2T) strongly related to the anaplastic grades were validated by ROC curve analysis (AUC >0.8). Based on the ssGSEA scores, the immune infiltration levels of Th2 cells were positively associated with anaplastic grade. qPCR assay showed that 9 hub genes were upregulated in RB cells, and UBE2T expressed remarkably high. CCK 8, EDU, and flow cytometry assays revealed that UBE2T silencing inhibited the proliferation of RB cells and incited apoptosis.

CONCLUSIONS

The increased infiltration of Th2 cells and upregulated expression of 9 hub genes predict a poor prognosis of RB. UBE2T can be a therapeutic target for RB treatment.

摘要

目的

本研究旨在筛选与视网膜母细胞瘤（RB）进展相关的去分化相关基因，并鉴定与预后不良相关的免疫细胞。

方法

从基因表达综合数据库（GEO）中获取视网膜和 RB 样本之间的差异表达基因（DEGs）。通过取验证集中的共表达基因、枢纽节点和 DEGs 的交集来筛选候选枢纽基因。通过接受者操作特征（ROC）和实时定量 PCR（qPCR）来鉴定枢纽基因。使用单样本基因集富集分析（ssGSEA）估计 RB 组织的免疫浸润水平。通过 CCK8、EDU 和流式细胞术检测 RB 细胞的功能。

结果

从 GEO 数据库中获得了 665 个涉及 RB 发生和进展的 DEGs。通过检查 43 个共表达基因和 63 个枢纽节点，筛选出 29 个候选枢纽基因。通过 ROC 曲线分析（AUC > 0.8）验证了 9 个与去分化程度密切相关的枢纽基因（CHEK1、EXO1、FANCI、GTSE1、MKI67、NCAPH、PRC1 和 UBE2T）。基于 ssGSEA 评分，Th2 细胞的免疫浸润水平与去分化程度呈正相关。qPCR 检测显示，9 个枢纽基因在 RB 细胞中上调，UBE2T 表达显著升高。CCK8、EDU 和流式细胞术检测表明，UBE2T 沉默抑制 RB 细胞增殖并诱导细胞凋亡。