BRIX1 promotes ribosome synthesis and enhances glycolysis by selected translation of GLUT1 in colorectal cancer

IF 3.2 4区医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Journal of Gene Medicine Pub Date : 2024-01-16 DOI:10.1002/jgm.3632

Chunhui Jiang, Longci Sun, Siyuan Wen, Yuan Tian, Chunjie Xu, Qing Xu, Hanbing Xue

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引用次数: 0

Abstract

Background

Ribosome biogenesis protein BRX1 homolog (BRIX1) is critically required for the synthesis of the 60S ribosome subunit. However, the role and mechanism of BRIX1 in colorectal cancer (CRC) remain unclear.

Methods

Kyoto Encyclopedia of Gene and Genome pathway and Gene Ontology analyses were used for bioinformatics analysis. The rRNA levels were detected in CRC tissues and cells. Nascent RNA synthesis was detected via cellular immunofluorescence. The correlation was analyzed between patient Positron Emission Tomography-Computed Tomography (PET–CT) values and their BRIX1 expression. The extracellular acidification rate (ECAR) and oxygen consumption rate were determined via live metabolic analyses. Polysome fractions were collected for BRIX1 mRNA used in translation. The orthotopic model and Cell Counting Kit-8 (CCK8) assay were used to assess BRIX1 function in CRC.

Results

BRIX1 is a core protein involved in ribosome-related pathway changes in CRC. Gene Ontology analysis showed that BRIX1 was primarily enriched in ribosome assembly and ribosome biogenesis pathways. In fresh CRC tissue, rRNA levels (5S, 5.8S, 18S and 28S) were higher in the BRIX1 high-expression group than in the BRIX1 low-expression group. Similarly, BRIX1 knockdown significantly decreased rRNA levels for 5S, 5.8S, 18S and 28S in CRC cells, whereas overexpression of BRIX1 significantly increased these levels. In addition, BRIX1 knockdown inhibited nascent RNA synthesis in CRC cells. In clinical data analysis, BRIX1 expression was related to the glucose uptake in PET–CT. BRIX1 knockdown significantly decreased the ECAR value, glucose uptake and lactic acid production in CRC cells, whereas BRIX1 overexpression significantly increased these. Furthermore, BRIX1 knockdown significantly decreased the protein expression of GLUT1, whereas BRIX1 overexpression significantly increased this; however, expression of BRIX1 mRNA was unaffected in either case. Blocking glycolysis by si-GLUT1 or galactose reversed BRIX1 promotion of glycolysis and cell proliferation in CRC cells.

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BRIX1 通过选择 GLUT1 的翻译促进结直肠癌中核糖体的合成并增强糖酵解作用

背景核糖体生物发生蛋白 BRX1 同源物（BRIX1）是合成 60S 核糖体亚基的关键所在。然而，BRIX1 在结直肠癌（CRC）中的作用和机制仍不清楚。方法采用京都基因百科全书和基因本体分析进行生物信息学分析。检测 CRC 组织和细胞中的 rRNA 水平。通过细胞免疫荧光检测新生 RNA 的合成。分析了患者正电子发射断层扫描（PET-CT）值与 BRIX1 表达之间的相关性。通过活体代谢分析确定了细胞外酸化率（ECAR）和耗氧量。收集多聚体碎片，以检测用于翻译的 BRIX1 mRNA。采用正位模型和细胞计数试剂盒-8（CCK8）检测法评估 BRIX1 在 CRC 中的功能。结果 BRIX1 是参与 CRC 核糖体相关通路变化的核心蛋白。基因本体分析表明，BRIX1主要富集于核糖体组装和核糖体生物发生通路。在新鲜的 CRC 组织中，BRIX1 高表达组的 rRNA 水平（5S、5.8S、18S 和 28S）高于 BRIX1 低表达组。同样，在 CRC 细胞中，BRIX1 基因敲除会显著降低 5S、5.8S、18S 和 28S 的 rRNA 水平，而过表达 BRIX1 则会显著提高这些水平。此外，BRIX1 基因敲除抑制了 CRC 细胞中新生 RNA 的合成。在临床数据分析中，BRIX1的表达与PET-CT的葡萄糖摄取有关。敲除 BRIX1 会明显降低 CRC 细胞的 ECAR 值、葡萄糖摄取量和乳酸生成量，而过表达 BRIX1 则会明显增加这些指标。此外，BRIX1敲除会明显降低GLUT1的蛋白表达，而BRIX1过表达则会明显提高GLUT1的蛋白表达。通过 si-GLUT1 或半乳糖阻断糖酵解可逆转 BRIX1 对 CRC 细胞糖酵解和细胞增殖的促进作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Gene Medicine 医学-生物工程与应用微生物

CiteScore

6.40

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： The aims and scope of The Journal of Gene Medicine include cutting-edge science of gene transfer and its applications in gene and cell therapy, genome editing with precision nucleases, epigenetic modifications of host genome by small molecules, siRNA, microRNA and other noncoding RNAs as therapeutic gene-modulating agents or targets, biomarkers for precision medicine, and gene-based prognostic/diagnostic studies. Key areas of interest are the design of novel synthetic and viral vectors, novel therapeutic nucleic acids such as mRNA, modified microRNAs and siRNAs, antagomirs, aptamers, antisense and exon-skipping agents, refined genome editing tools using nucleic acid /protein combinations, physically or biologically targeted delivery and gene modulation, ex vivo or in vivo pharmacological studies including animal models, and human clinical trials. Papers presenting research into the mechanisms underlying transfer and action of gene medicines, the application of the new technologies for stem cell modification or nucleic acid based vaccines, the identification of new genetic or epigenetic variations as biomarkers to direct precision medicine, and the preclinical/clinical development of gene/expression signatures indicative of diagnosis or predictive of prognosis are also encouraged.