Zhang Jidong, You Qiuxiang, Wang Yutao, Ji Jianwen
Center for Neurological Diseases, The Third Affiliated Hospital of Chongqing Medical University, Chongqing, 401120, People's Republic of China.
Biologics. 2024 May 23;18:129-142. doi: 10.2147/BTT.S454058. eCollection 2024.
The main purpose of this paper is to explore the interaction between GAS5 and miR-135b-5p to understand their function in the metastasis, invasion, and proliferation of glioma. This may provide new ideas for the pathogenesis and treatment of glioma.
Western blotting assays and RT‑qPCR were employed to investigate the expression of related genes in glioma tissues or cell lines. CCK-8 was used to examine the impact of GAS5 on cell viability. Motile activities were adopted by the transwell and wound healing experiments. A double luciferase experiment was performed to elucidate transcriptional regulation.
GAS5 showed low expression in glioma cells and tissues, and up-regulation of GAS5 could depress the invasion, proliferation, and metastasis of glioma. GAS5 negatively regulates miR-135b-5p, which can counteract the cellular effects caused by GAS5. APC was the target of miR-135b-5p, and GAS5 can regulate the expression of APC by sponging miR-135b-5p. APC overexpression reversed the effects of miR-135b-5p promotion on glioma cells, while miR-135b-5p has the opposite function. As a downstream target gene of GAS5, miR-135b-5p was negatively regulated by GAS5. The restoration of miR-135b-5p can remarkably reverse the impact of GAS5 on glioma cells. In addition, GAS5 increased the expression of APC in glioma cells by inhibiting miR-135b-5p.
GAS5 increased APC expression by restraining miR-135b-5p and partially blocked the progression of glioma, suggesting that it could be an advantageous therapeutic target for glioma intervention.
本文的主要目的是探讨GAS5与miR-135b-5p之间的相互作用,以了解它们在胶质瘤转移、侵袭和增殖中的作用。这可能为胶质瘤的发病机制和治疗提供新思路。
采用蛋白质免疫印迹法和逆转录定量聚合酶链反应(RT-qPCR)检测胶质瘤组织或细胞系中相关基因的表达。使用细胞计数试剂盒-8(CCK-8)检测GAS5对细胞活力的影响。通过Transwell实验和伤口愈合实验检测细胞的运动活性。进行双荧光素酶实验以阐明转录调控。
GAS5在胶质瘤细胞和组织中表达较低,上调GAS5可抑制胶质瘤的侵袭、增殖和转移。GAS5负向调节miR-135b-5p,而miR-135b-5p可抵消GAS5引起的细胞效应。腺瘤性息肉病 coli(APC)是miR-135b-5p的靶标,GAS5可通过吸附miR-135b-5p来调节APC的表达。APC过表达可逆转miR-135b-5p促进胶质瘤细胞的作用,而miR-135b-5p则具有相反的功能。作为GAS5的下游靶基因,miR-135b-5p受到GAS5的负向调节。miR-135b-5p的恢复可显著逆转GAS5对胶质瘤细胞的影响。此外,GAS5通过抑制miR-135b-5p增加胶质瘤细胞中APC的表达。
GAS5通过抑制miR-135b-5p增加APC表达,部分阻断胶质瘤的进展,提示其可能是胶质瘤干预的有利治疗靶点。