一种新型长链非编码RNA lncRNA - 4045通过影响AKR1B10的表达促进肝细胞癌进展。

A Novel lncRNA lncRNA-4045 Promotes the Progression of Hepatocellular Carcinoma by Affecting the Expression of AKR1B10.

作者信息

Tan Chao, Zeng Xi, Guo Xuefeng, Mo Meile, Ma Xiaoyun, Liu Bihu, Liu Shun, Zeng Xiaoyun, Huang Dongping, Qiu Xiaoqiang

机构信息

Department of Epidemiology and Health Statistics, School of Public Health, Guilin Medical University, Zhiyuan Road, Guilin, 541199, Guangxi, People's Republic of China.

Department of Occupational and Environmental Health, School of Public Health, Guilin Medical University, Zhiyuan Road, Guilin, 541199, Guangxi, People's Republic of China.

出版信息

Dig Dis Sci. 2024 Jul;69(7):2502-2521. doi: 10.1007/s10620-024-08383-z. Epub 2024 Apr 25.

DOI:10.1007/s10620-024-08383-z

PMID:38662158

Abstract

BACKGROUND

Long noncoding RNAs (lncRNAs) have been shown to be related to the occurrence and development of a variety of cancers including hepatocellular carcinoma (HCC). However, a large number of potential HCC-related lncRNAs remain undiscovered and are yet to be fully understood.

METHODS

Differentially expressed lncRNAs were first obtained from the tumor tissues and adjacent normal tissues of five HCC patients using high-throughput microarray chips. Then the expression levels of 10 differentially expressed lncRNAs were verified in 50 pairs of tissue samples from patients with HCC by quantitative real-time PCR (qRT-PCR). The oncogenic effects of lncRNA-4045 (ENST00000524045.6) in HCC cell lines were verified through a series of in vitro experiments including CCK-8 assay, plate clone formation assay, transwell assay, scratch assay, and flow cytometry. Subsequently, the potential target genes of lncRNA-4045 were predicted by bioinformatics analysis, fluorescence in situ hybridization assay, and RNA sequencing. The mechanism of lncRNA-4045 in HCC was explored by WB assay as well as rescue and enhancement experiments.

RESULTS

The results from microarray chips showed 1,708 lncRNAs to have been significantly upregulated and 2725 lncRNAs to have been significantly downregulated in HCC tissues. Via validation in 50 HCC patients, a novel lncRNA lncRNA-4045 was found significantly upregulated in HCC tissues. Additionally, a series of in vitro experiments showed that lncRNA-4045 promoted the proliferation, invasion, and migration of HCC cell lines, and inhibited the apoptosis of HCC cell lines. The results of qRT-PCR in HCC tissues showed that the expression levels of AKR1B10 were significantly positively correlated with lncRNA-4045. LncRNA-4045 knockdown significantly down-regulated AKR1B10 protein expression, and overexpression of lncRNA-4045 led to significant up-regulation of AKR1B10 protein in HCC cell lines. Lastly, down-regulation of AKR1B10 could partially eliminate the enhancement of cell proliferation induced by lncRNA-4045 overexpression, while up-regulation of AKR1B10 was shown to enhance those effects.

CONCLUSION

LncRNA-4045 may promote HCC via enhancement of the expression of AKR1B10 protein.

摘要

背景

长链非编码RNA（lncRNAs）已被证明与包括肝细胞癌（HCC）在内的多种癌症的发生和发展有关。然而，大量潜在的与HCC相关的lncRNAs仍未被发现，有待充分了解。

方法

首先使用高通量微阵列芯片从5例HCC患者的肿瘤组织和相邻正常组织中获得差异表达的lncRNAs。然后通过定量实时PCR（qRT-PCR）在50对HCC患者组织样本中验证10种差异表达lncRNAs的表达水平。通过一系列体外实验，包括CCK-8检测、平板克隆形成检测、Transwell检测、划痕检测和流式细胞术，验证lncRNA-4045（ENST00000524045.6）在HCC细胞系中的致癌作用。随后，通过生物信息学分析、荧光原位杂交检测和RNA测序预测lncRNA-4045的潜在靶基因。通过蛋白质免疫印迹法（WB）以及拯救和增强实验探索lncRNA-4045在HCC中的作用机制。

结果

微阵列芯片结果显示，HCC组织中有1708种lncRNAs显著上调，2725种lncRNAs显著下调。通过对50例HCC患者的验证，发现一种新型lncRNA lncRNA-4045在HCC组织中显著上调。此外，一系列体外实验表明，lncRNA-4能促进HCC细胞系的增殖、侵袭和迁移，并抑制HCC细胞系的凋亡。HCC组织中qRT-PCR结果显示，AKR1B10的表达水平与lncRNA-4045显著正相关。lncRNA-4045敲低显著下调AKR1B10蛋白表达，lncRNA-4045过表达导致HCC细胞系中AKR1B10蛋白显著上调。最后，下调AKR1B10可部分消除lncRNA-4045过表达诱导的细胞增殖增强，而上调AKR1B10则增强这些作用。