Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, School of Life Science, Jiangsu Normal University, Xuzhou, China.
College of Health Sciences, Jiangsu Normal University, Xuzhou, Jiangsu, China.
J Cell Mol Med. 2018 Oct;22(10):4963-4974. doi: 10.1111/jcmm.13760. Epub 2018 Jul 19.
This study was designed to explore the relationship between miR-1275 and SERPINE1 and its effects on glioma cell proliferation, migration, invasion and apoptosis. Differentially expressed miRNAs and mRNAs in glioma tissues were screened out by bioinformatic analysis. Dual-luciferase reporter gene assay was used to validate the targeted relationship between miR-1275 and SERPINE1. qRT-PCR was used to detect the expression of miR-1275 and SERPINE1 in glioma tissues. The expressions of SERPINE1 and p53 pathway-related proteins in glioma cells were detected by western blot. Glioma cell proliferation, apoptosis, migration and invasion were respectively detected by CCK-8 assay, flow cytometry, wound healing assay and transwell assay. Tumour xenograft model was developed to study the influence of miR-1275 and SERPINE1 on glioma growth in vivo. The results of microarray analysis, qRT-PCR and western blot showed that miR-1275 was low-expressed while SERPINE1 was high-expressed in glioma. Dual-luciferase assay showed that miR-1275 could bind to SERPINE1. Overexpression of miR-1275 could promote the p53 pathway-related proteins' expression. Highly expressed miR-1275 could repress the migration, proliferation and invasion of glioma cells while highly expressed SERPINE1 had inverse effects. Tumour xenograft showed that up-regulated miR-1275 or down-regulated SERPINE1 could repress glioma growth in vivo. Up-regulation of miR-1275 activated p53 signalling pathway via regulating SERPINE1 and therefore suppressed glioma cell proliferation, invasion and migration, whereas promoted cell apoptosis.
本研究旨在探讨 miR-1275 与 SERPINE1 之间的关系及其对神经胶质瘤细胞增殖、迁移、侵袭和凋亡的影响。通过生物信息学分析筛选出神经胶质瘤组织中差异表达的 miRNAs 和 mRNAs。双荧光素酶报告基因实验验证 miR-1275 与 SERPINE1 的靶向关系。qRT-PCR 检测神经胶质瘤组织中 miR-1275 和 SERPINE1 的表达。Western blot 检测神经胶质瘤细胞中 SERPINE1 和 p53 通路相关蛋白的表达。CCK-8 法检测神经胶质瘤细胞增殖，流式细胞术检测细胞凋亡，划痕愈合实验和 Transwell 实验分别检测细胞迁移和侵袭。建立肿瘤异种移植模型，研究 miR-1275 和 SERPINE1 对神经胶质瘤体内生长的影响。微阵列分析、qRT-PCR 和 Western blot 结果显示，miR-1275 在神经胶质瘤中低表达，而 SERPINE1 高表达。双荧光素酶实验表明，miR-1275 可以与 SERPINE1 结合。过表达 miR-1275 可以促进 p53 通路相关蛋白的表达。高表达 miR-1275 可以抑制神经胶质瘤细胞的迁移、增殖和侵袭，而高表达 SERPINE1 则具有相反的作用。肿瘤异种移植显示，上调 miR-1275 或下调 SERPINE1 可以抑制体内神经胶质瘤的生长。上调 miR-1275 通过调节 SERPINE1 激活 p53 信号通路，从而抑制神经胶质瘤细胞的增殖、侵袭和迁移，同时促进细胞凋亡。
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