抑制长链非编码 RNA NEAT1 通过降低连接蛋白 43 的表达增加胶质母细胞瘤对 TMZ 的反应。

Inhibiting lncRNA NEAT1 Increases Glioblastoma Response to TMZ by Reducing Connexin 43 Expression.

机构信息

Guangxi Key Laboratory of Traditional Chinese Medicine Quality Standards, Research Center of Traditional Chinese Medicine and Ethnic Medicine, Guangxi Institute of Chinese Medicine and Pharmaceutical Science, Nanning, China.

Pharmaceutical College, Guangxi Medical University, Nanning, China.

出版信息

Cancer Rep (Hoboken). 2024 Oct;7(10):e70031. doi: 10.1002/cnr2.70031.

DOI:10.1002/cnr2.70031

PMID:39453684

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11505515/

Abstract

OBJECTIVES

Glioblastoma multiforme (GBM) is considered the most assailant subtype of gliomas, presenting a formidable obstacle because of its inherent resistance to temozolomide (TMZ). This study aimed to characterize the function of lncRNA NEAT1 in facilitating the advancement of gliomas.

METHODS

The expression level of NEAT1 in glioma tissues and cells was detected by qRT-PCR. RNA interference experiment, cell proliferation assay, FITC/PI detection assay, immunoblotting, bioinformatics prediction, a double luciferase reporter gene assay, RNA immunoprecipitation (RIP) assay, SLDT assay and correlation analysis of clinical samples were performed to explore the regulatory effects of NEAT1, miR-454-3p and Cx43 and their role in malignant progression of GBM. The role of NEAT1 in vivo was investigated by an intracranial tumor formation experiment in mice.

RESULTS

The results showed that recurring gliomas displayed elevated levels of NEAT1 compared to primary gliomas. The suppression of NEAT1 led to a restoration of sensitivity in GBM cells to TMZ. NEAT1 functioned as a competitive endogenous RNA against miR-454-3p. Connexin 43 was identified as a miR-454-3p target. NEAT1 was found to regulate gap junctional intercellular communication by modulating Connexin 43, thereby impacting the response of GBM cells to TMZ chemotherapy. Downregulation of NEAT1 resulted in enhanced chemosensitivity to TMZ and extended the survival of mice.

CONCLUSIONS

Overall, these results indicated that the NEAT1/miR-454-3p/Connexin 43 pathway influences GBM cell response to TMZ and could offer a potential new strategy for treating GBM.

摘要

目的

多形性胶质母细胞瘤（GBM）被认为是最具侵袭性的神经胶质瘤亚型，由于其对替莫唑胺（TMZ）的固有耐药性，成为治疗的巨大障碍。本研究旨在研究 lncRNA NEAT1 在促进神经胶质瘤进展中的作用。

方法

采用 qRT-PCR 检测神经胶质瘤组织和细胞中 NEAT1 的表达水平。通过 RNA 干扰实验、细胞增殖实验、FITC/PI 检测实验、免疫印迹实验、生物信息学预测、双荧光素酶报告基因实验、RNA 免疫沉淀（RIP）实验、SLDT 实验以及临床样本的相关性分析，探讨 NEAT1、miR-454-3p 和 Cx43 的调控作用及其在 GBM 恶性进展中的作用。通过在小鼠颅内肿瘤形成实验中研究 NEAT1 的体内作用。

结果

结果表明，复发性神经胶质瘤中 NEAT1 的水平高于原发性神经胶质瘤。抑制 NEAT1 可使 GBM 细胞对 TMZ 恢复敏感性。NEAT1 作为竞争性内源性 RNA 与 miR-454-3p 相互作用。连接蛋白 43 被鉴定为 miR-454-3p 的靶基因。通过调节 Connexin 43，NEAT1 被发现调节缝隙连接细胞间通讯，从而影响 GBM 细胞对 TMZ 化疗的反应。下调 NEAT1 可增强对 TMZ 的化疗敏感性并延长小鼠的存活时间。