miRNA-383-5p 通过靶向 VEGFA 作用于 Akt/mTOR 信号通路在胶质瘤中发挥抑癌基因作用。

mirna-383-5p Functions as an Anti-oncogene in Glioma through the Akt/mTOR Signaling Pathway by Targeting VEGFA.

机构信息

Department of Neurosurgery, Hunan Cancer Hospital and the Affiliated Cancer Hospital of Xiangya School of Medicine, Central South University, No. 283 Tongzipo Road, Yuelu District, Changsha, 410006, Hunan, China.

Department of Neurology, Changsha Central Hospital, University of South China, No.161 Shaoshan Road, Yuhua District, Changsha, 410007, Hunan, China.

出版信息

Curr Cancer Drug Targets. 2024;24(4):463-475. doi: 10.2174/1568009623666230817102104.

DOI:10.2174/1568009623666230817102104

PMID:37592783

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10964077/

Abstract

BACKGROUND

Previously, we have screened 59 differentially expressed miRNAs and 419 mRNAs in the glioblastoma samples that have been compared to the peritumoral tissues using bioinformatics analyses, which included miRNA-383-5p and vascular endothelial growth factor A (VEGFA). miRNA-383-5p and VEGFA/Akt/mTOR pathway play important regulatory roles in the malignant biological behavior of glioma.

METHODS

Glioma cell lines, U87 and U251, were collected for in vitro experiments. miRNA-383-5p and VEGFA expression levels were detected with qRT-PCR and WB. The protein expressions of Akt, mTOR, and VEGFR in U87 and U251 were detected with WB. The effect of miRNA-383-5p on the VEGFA activity was verified by dual-luciferase reporter assay. CCK-8 was used to examine the U87 and U251 cells' inhibition. Flow cytometry and transwell assays were used to detect cell apoptosis and invasion, respectively.

RESULTS

Our research data indicated overexpression of miRNA-383-5p to suppress malignant biological behavior, which was manifested as promoting the apoptosis of U87 and U251 cells and inhibiting invasion, proliferation, and metastasis. VEGFA is one of the downstream target genes of miRNA-383- 5p. miRNA-383-5p could inhibit the expression of VEGFA and Akt/mTOR signaling pathways. Overexpression of VEGFA can reverse the inhibitory effect of miRNA-383-5p and reactivate the Akt/mTOR signaling pathway.

CONCLUSION

Our results indicate that miRNA-383-5p functions as an anti-oncogene by inhibiting the VEGFA/Akt/mTOR signaling pathway in glioma cells. These data provide potential therapeutic targets for glioblastoma.

摘要

背景

此前，我们通过生物信息学分析筛选出胶质瘤样本中 59 个差异表达的 miRNA 和 419 个 mRNA，与肿瘤周围组织进行比较，其中包括 miRNA-383-5p 和血管内皮生长因子 A（VEGFA）。miRNA-383-5p 和 VEGFA/Akt/mTOR 通路在胶质瘤的恶性生物学行为中发挥重要的调节作用。

方法

收集胶质瘤细胞系 U87 和 U251 进行体外实验。qRT-PCR 和 WB 检测 miRNA-383-5p 和 VEGFA 的表达水平。WB 检测 U87 和 U251 中 Akt、mTOR 和 VEGFR 的蛋白表达。双荧光素酶报告实验验证 miRNA-383-5p 对 VEGFA 活性的影响。CCK-8 检测 U87 和 U251 细胞的抑制作用。流式细胞术和 Transwell 实验分别检测细胞凋亡和侵袭。

结果

我们的研究数据表明，miRNA-383-5p 的过表达抑制恶性生物学行为，表现为促进 U87 和 U251 细胞的凋亡，抑制侵袭、增殖和转移。VEGFA 是 miRNA-383-5p 的下游靶基因之一。miRNA-383-5p 可以抑制 VEGFA 和 Akt/mTOR 信号通路的表达。过表达 VEGFA 可以逆转 miRNA-383-5p 的抑制作用并重新激活 Akt/mTOR 信号通路。