[UBE2S高表达通过增加癌细胞干性促进肝细胞癌进展]

[High expression of UBE2S promotes progression of hepatocellular carcinoma by increasing cancer cell stemness].

作者信息

Chen H, Li Z, Wang M, Lu L, Tang Q, Luo L

机构信息

Postdoctoral Research Station of Clinical Medicine, Jinan University, Guangzhou 510632, China.

Graduate School, Youjiang Medical University for Nationalities, Baise 533000, China.

出版信息

Nan Fang Yi Ke Da Xue Xue Bao. 2024 Mar 20;44(3):455-464. doi: 10.12122/j.issn.1673-4254.2024.03.06.

DOI:10.12122/j.issn.1673-4254.2024.03.06

PMID:38597436

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11006698/

Abstract

OBJECTIVE

To investigate the expression of the ubiquitination enzyme UBE2S in different cell types in hepatocellular carcinoma (HCC) microenvironment and its impact on proliferation and stemness of HCC cells.

METHODS

TCGA and CPTAC database were used to analyze the transcriptional and promoter methylation levels and protein expressions of UBE2S in HCC. Specific expression patterns of UBE2S, intercellular communication and key transcription factors in different cell types were analyzed based on single-cell sequencing data from TISCH website. We further examined UBE2S expressions in clinical samples of HCC tissues, HCC cells and T cells using immunohistochemistry and immunofluorescence staining. We also tested the effects of UBE2S knockdown on stemness of HCC-LM3 and HepG2 cells using clone formation experiments and sphere formation assay.

RESULTS

Analysis based on TCGA database suggested significant overexpression of UBE2S in both paired and non-paired tumor tissues ( < 0.001), and its transcriptional level increased with tumor grades. The methylation level of UBE2S promoter was significantly decreased in HCC ( < 0.001), and its transcription level increased obviously in HCC with TP53 mutation ( < 0.001). Analysis of CPTAC database also demonstrated overexpression of UBE2S protein in HCC tissues ( < 0.001). Three prognostic models suggested that HCC patients with high UBE2S expression had poorer prognosis ( < 0.001). Single-cell sequencing data analysis revealed high expressions of UBE2S in T cells and high intensities of interaction between endothelial cells, epithelial cells and fibroblasts in HCC microenvironment. Immunohistochemistry and immunofluorescence staining demonstrated high UBE2S expressions in clinical samples of HCC tissues, HCC cells and T cells. In HCC-LM3 and HepG2 cells, UBE2S knockdown significantly inhibited cell clone formation and tumor sphere formation ( < 0.05).

CONCLUSION

UBE2S is highly expressed in T cells in HCC microenvironment in close correlation with a poor prognosis. High UBE2S expression promotes the stemness of HCC cells.

摘要

目的

研究泛素化酶UBE2S在肝细胞癌（HCC）微环境中不同细胞类型中的表达及其对HCC细胞增殖和干性的影响。

方法

利用TCGA和CPTAC数据库分析HCC中UBE2S的转录水平、启动子甲基化水平及蛋白表达。基于TISCH网站的单细胞测序数据，分析UBE2S在不同细胞类型中的特异性表达模式、细胞间通讯及关键转录因子。我们还采用免疫组织化学和免疫荧光染色法检测HCC组织、HCC细胞和T细胞临床样本中UBE2S的表达。我们还通过克隆形成实验和球体形成试验检测了UBE2S敲低对HCC-LM3和HepG2细胞干性的影响。

结果

基于TCGA数据库的分析表明，UBE2S在配对和未配对的肿瘤组织中均显著过表达（<0.001），且其转录水平随肿瘤分级增加。HCC中UBE2S启动子的甲基化水平显著降低（<0.001），在TP53突变的HCC中其转录水平明显升高（<0.001）。CPTAC数据库分析也显示HCC组织中UBE2S蛋白过表达（<0.001）。三种预后模型表明，UBE2S高表达的HCC患者预后较差（<0.001）。单细胞测序数据分析显示，T细胞中UBE2S高表达，且HCC微环境中内皮细胞、上皮细胞和成纤维细胞之间的相互作用强度较高。免疫组织化学和免疫荧光染色显示，HCC组织、HCC细胞和T细胞临床样本中UBE2S高表达。在HCC-LM3和HepG2细胞中，UBE2S敲低显著抑制细胞克隆形成和肿瘤球体形成（<0.05）。