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lncRNA CHRM3-AS2表达沉默通过靶向microRNA-370-5p/KLF4发挥抗胶质瘤肿瘤作用。

Silencing of lncRNA CHRM3-AS2 Expression Exerts Anti-Tumour Effects Against Glioma Targeting microRNA-370-5p/KLF4.

作者信息

Wang Dong, Chen Qiang, Liu Jun, Liao Yuqing, Jiang Qiuhua

机构信息

Department of Neurosurgery, Ganzhou People's Hospital, Ganzhou, China.

出版信息

Front Oncol. 2022 Mar 11;12:856381. doi: 10.3389/fonc.2022.856381. eCollection 2022.

DOI:10.3389/fonc.2022.856381
PMID:35359381
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8962832/
Abstract

OBJECTIVES

Long non-coding RNAs (lncRNAs) are key regulators involved in the progression of glioma, and many functional lncRNAs are yet to be identified. This study aimed to explore the function of CHRM3-AS2, a rarely reported lncRNA, in glioma, as well as the underlying mechanisms involving miR-370-5p/KLF4.

METHODS

Differentially expressed RNAs (DERs) were screened from two gene expression profiles of glioblastoma (GBM). Fluorescence hybridisation was performed to determine the subcellular localisation of CHRM3-AS2. Cell viability, colony formation, apoptosis, migration, and invasion were evaluated using cell counting kit-8, colony counts, flow cytometry, wound healing, and Transwell assays, respectively. mRNA and protein expression of specific genes were measured using quantitative real-time polymerase chain reaction and western blotting, respectively. Dual luciferase reporter gene, RNA immunoprecipitation, and RNA pull-down assays were performed to identify the target relationships. A mouse xenograft model was established for validation.

RESULTS

CHRM3-AS2 was screened as a prognosis-associated DER in GBM. CHRM3-AS2 expression was up-regulated in glioma cells, and CHRM3-AS2 was localised in the cytoplasm. Silencing of CHRM3-AS2 expression inhibited cell viability, colony formation, migration, and invasion and promoted apoptosis of U251 and SHG-44 cells. In addition, CHRM3-AS2 targeted miR-370-5p/KLF4 in glioma cells. The anti-tumour effect of CHRM3-AS2 silencing was weakened by miR-370-5p silencing or KLF4 overexpression. , silencing of CHRM3-AS2 expression inhibited tumour growth and Ki67 expression in mice. Overexpression of KLF4 also weakened the anti-tumour effect of CHRM3-AS2 silencing in mice.

CONCLUSIONS

Silencing of CHRM3-AS2 expression inhibited the malignant progression of glioma by regulating miR-370-5p/KLF4 expression.

摘要

目的

长链非编码RNA（lncRNAs）是参与胶质瘤进展的关键调节因子，许多功能性lncRNAs尚待鉴定。本研究旨在探讨一种报道较少的lncRNA CHRM3-AS2在胶质瘤中的功能，以及涉及miR-370-5p/KLF4的潜在机制。

方法

从胶质母细胞瘤（GBM）的两个基因表达谱中筛选差异表达RNA（DERs）。进行荧光杂交以确定CHRM3-AS2的亚细胞定位。分别使用细胞计数试剂盒-8、集落计数、流式细胞术、伤口愈合和Transwell实验评估细胞活力、集落形成、凋亡、迁移和侵袭。分别使用定量实时聚合酶链反应和蛋白质印迹法测量特定基因的mRNA和蛋白质表达。进行双荧光素酶报告基因、RNA免疫沉淀和RNA下拉实验以鉴定靶标关系。建立小鼠异种移植模型进行验证。

结果

CHRM3-AS2被筛选为GBM中与预后相关的DER。CHRM3-AS2在胶质瘤细胞中表达上调，且CHRM3-AS2定位于细胞质中。沉默CHRM3-AS2表达可抑制U251和SHG-44细胞活力、集落形成、迁移和侵袭，并促进其凋亡。此外，CHRM3-AS2在胶质瘤细胞中靶向miR-370-5p/KLF4。沉默miR-370-5p或过表达KLF4可削弱CHRM3-AS2沉默的抗肿瘤作用。此外，沉默CHRM3-AS2表达可抑制小鼠肿瘤生长和Ki67表达。过表达KLF4也削弱了CHRM3-AS2沉默在小鼠中的抗肿瘤作用。

结论

沉默CHRM3-AS2表达通过调节miR-370-5p/KLF4表达抑制胶质瘤的恶性进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4665/8962832/0f2027e93108/fonc-12-856381-g008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4665/8962832/9084339b57b8/fonc-12-856381-g005.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4665/8962832/0f2027e93108/fonc-12-856381-g008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4665/8962832/9084339b57b8/fonc-12-856381-g005.jpg
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