Neurosurgery Department, Nanyang Second General Hospital, China.
Department of Rheumatology and Immunology, Affiliated Hospital of Southwest Medical University, China.
Brain Res Bull. 2021 Aug;173:82-96. doi: 10.1016/j.brainresbull.2021.05.010. Epub 2021 May 13.
A strong relationship between long intergenic non-protein coding RNA 511 (LINC00511) and glioma has been previously reported but the mechanism of LINC00511 in glioma is yet to be determined. This study examined the mechanism of LINC00511 in glioma.
The expression of LINC00511 in glioma was determined by bioinformatics analysis and real-time quantitative PCR (RT-qPCR) analysis. The target relationship between genes was predicted by starBase, TargetScan, and was verified by dual-luciferase. Subsequently, siRNA targeting LINC00511 (siLINC00511) and miR-15a-5p mimic were transfected into glioma cells to examine the effect on biological characteristics using cell counting kit-8, clone formation, flow cytometry, wound-healing, and transwell. MiR-15a-5p inhibitor and AEBP1 were used for in vitro rescue experiments, and tumorigenesis assay and immunohistochemical assays were performed for in vivo experiments. Epithelial-mesenchymal transition (EMT) and p65 phosphorylation were examined by Western blot.
LINC00511 was predicted and verified to be up-regulated in glioma. SiLINC00511 suppressed cell viability, proliferation, migration and invasion, accelerated apoptosis of glioma cells. Mechanically, siLINC00511 promoted E-cadherin expression but suppressed N-cadherin and Snail expressions. MiR-15a-5p bound to LINC00511, and miR-15a-5p inhibitor partially reversed the effect and regulation of siLINC00511 on glioma cells. AEBP1, a target gene of miR-15a-5p, could activate p65 phosphorylation to promote EMT protein expression and partially reverse the inhibitory effect of miR-15a-5p mimic on the malignant phenotype of glioma cells. SiLINC00511 inhibited tumor growth, down-regulated miR-15a-5p expression and up-regulated AEBP1 and Ki67 expressions in vivo.
LINC00511 knockdown inhibits glioma cell progression via miR-15a-5p/AEBP1 axis.
长链非编码 RNA 511（LINC00511）与神经胶质瘤之间存在很强的关系，但 LINC00511 在神经胶质瘤中的作用机制尚不清楚。本研究探讨了 LINC00511 在神经胶质瘤中的作用机制。
通过生物信息学分析和实时定量 PCR（RT-qPCR）分析检测神经胶质瘤中 LINC00511 的表达。通过 starBase、TargetScan 预测基因之间的靶关系，并通过双荧光素酶报告基因进行验证。随后，将靶向 LINC00511 的 siRNA（siLINC00511）和 miR-15a-5p 模拟物转染入神经胶质瘤细胞中，通过细胞计数试剂盒-8、集落形成、流式细胞术、划痕愈合和 Transwell 实验检测其对细胞生物学特性的影响。用 miR-15a-5p 抑制剂和 AEBP1 进行体外挽救实验，进行体内实验的肿瘤发生测定和免疫组化分析。通过 Western blot 检测上皮间质转化（EMT）和 p65 磷酸化。
预测并验证 LINC00511 在神经胶质瘤中上调。SiLINC00511 抑制神经胶质瘤细胞活力、增殖、迁移和侵袭，促进细胞凋亡。机制上，siLINC00511 促进 E-钙黏蛋白表达，抑制 N-钙黏蛋白和 Snail 表达。MiR-15a-5p 与 LINC00511 结合，miR-15a-5p 抑制剂部分逆转了 siLINC00511 对神经胶质瘤细胞的作用和调控。AEBP1 是 miR-15a-5p 的靶基因，可激活 p65 磷酸化，促进 EMT 蛋白表达，并部分逆转 miR-15a-5p 模拟物对神经胶质瘤细胞恶性表型的抑制作用。体内实验中，siLINC00511 抑制肿瘤生长，下调 miR-15a-5p 表达，上调 AEBP1 和 Ki67 表达。
LINC00511 敲低通过 miR-15a-5p/AEBP1 轴抑制神经胶质瘤细胞的进展。
