由BHLHE40诱导的长链非编码RNA NEAT1激活Wnt/β-连环蛋白信号通路并促进结直肠癌进展。

Long non-coding RNA NEAT1 induced by BHLHE40 activates Wnt/β-catenin signaling and potentiates colorectal cancer progression.

作者信息

Ji Anlong, Li Hui, Fu Xiangwei, Zhang Yourong, Liu Yanhe

机构信息

Department of General Surgery, The Second Affiliated Hospital of Hainan Medical University, No. 368, Yehai Avenue, Haikou, 570216, Hainan, People's Republic of China.

Department of Geriatrics, The Second Affiliated Hospital of Hainan Medical University, No. 368, Yehai Avenue, Haikou, 570216, Hainan, People's Republic of China.

出版信息

Cell Div. 2024 Aug 4;19(1):25. doi: 10.1186/s13008-024-00129-7.

DOI:10.1186/s13008-024-00129-7

PMID:39098910

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11299305/

Abstract

BACKGROUND

Nuclear-enriched abundant transcript 1 (NEAT1), a long noncoding RNA (lncRNA), has been implicated in the colorectal cancer (CRC) progression. However, its upstream mechanism has not been well studied. In the present study, the functions and mechanisms of NEAT1 in CRC were investigated.

METHODS

The NEAT1 expression in CRC tissues and CRC cells was analyzed by RT-qPCR. The genes co-expressed with NEAT1 in CRC were obtained from UALCAN, which were intersected with the transcription factors targeting NEAT1 from hTFtarget. Dual-luciferase assay, RT-qPCR, and ChIP were conducted to analyze the transcriptional regulatory relationship between BHLHE40 and NEAT1. LoVo and HCT-15 cells knocking down BHLHE40 and overexpressing NEAT1 were subjected to MTT, Transwell, Western blot, and flow cytometry to examine the malignant aggressiveness of CRC cells. The effects of knocking down BHLHE40 and overexpressing NEAT1 on tumor and lung metastasis were investigated in mice using HE and immunohistochemical analyses.

RESULTS

NEAT1 and BHLHE40 were significantly overexpressed in CRC tissues and cells. BHLHE40 has a binding relationship with the NEAT1 promoter. Knockdown of BHLHE40 resulted in a reverted malignant phenotype in vitro and slowed tumor growth and metastasis dissemination in vivo, which were reversed by NEAT1 overexpression. Overexpression of BHLHE40 increased Wnt/β-catenin pathway activity, but knockdown of NEAT1 decreased Wnt/β-catenin pathway activity.

CONCLUSIONS

BHLHE40 mediates the transcriptional activation of NEAT1, which activates the Wnt/β-catenin pathway and promotes the CRC progression.

摘要

背景

富含核仁的丰富转录本1（NEAT1）是一种长链非编码RNA（lncRNA），与结直肠癌（CRC）进展有关。然而，其上游机制尚未得到充分研究。在本研究中，对NEAT1在CRC中的功能和机制进行了研究。

方法

通过RT-qPCR分析CRC组织和CRC细胞中NEAT1的表达。从UALCAN获得与CRC中NEAT1共表达的基因，并与hTFtarget中靶向NEAT1的转录因子进行交叉分析。进行双荧光素酶测定、RT-qPCR和染色质免疫沉淀分析BHLHE40与NEAT1之间的转录调控关系。对敲低BHLHE40并过表达NEAT1的LoVo和HCT-15细胞进行MTT、Transwell、蛋白质免疫印迹和流式细胞术检测，以研究CRC细胞的恶性侵袭性。使用苏木精-伊红（HE）和免疫组织化学分析，研究敲低BHLHE40和过表达NEAT1对小鼠肿瘤和肺转移的影响。

结果

NEAT1和BHLHE40在CRC组织和细胞中显著过表达。BHLHE40与NEAT1启动子存在结合关系。敲低BHLHE40可在体外逆转恶性表型，并在体内减缓肿瘤生长和转移扩散，而NEAT1过表达可逆转这些作用。BHLHE40的过表达增加了Wnt/β-连环蛋白信号通路的活性，但敲低NEAT1则降低了Wnt/β-连环蛋白信号通路的活性。